@marroweditionénotes

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BIOCHEMISTRY OF FED STATE
Fed state/absorptive phase :
• fiso called as post prandial state.

• Starts within 3-4 hours of food intake.

• Digeston § absorption of food fares place.

• Plasma level of glucose, amino acids, Fatty acids 5 fern ‘yee level increases.

• chylomicrons which canny triacy gycerol alo increases.

Insulin secretion 002s

Hormone of fed state : Insulin.
sulin starts 1 rie when Hood glucose level» 39 rmal/ Lor maf
Glucose ronsparted fo beta cel of pancreas through
bly Low ait br Gm yroweditionénotes once see te beta cells encase | econinase cuense
pesghote
Pyrite > arp
ATP/ROP ratio increases which results in closure of ATP Sensitive K' channels.
Resulting in depolarisation of membrane.
Opening of voitage goted caleum chamels opens, resulting in ca* influx.
Calcium stimulote secretory vesicles carrying nul, resulting in ts secretion.
C peptide co-secreted along wih insulin.
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BIOCHEMISTRY OF FASTING STATE
Stages of post absorptive state 00:00:55,
Early fasting : metabolic changes + + 4 -lehours without food intake. * Source oF blood
glucose : siyeogenclysis. + end product of glyeogenolyss is glucose (hepatic aiyregensysis).
. Ss ans aes ait for motes « bea.
souree of blood glucose in this state. . Weiecty ule provides blood use. + In le-48 hours,
glycogen stores are depleted.
Fasting : Metabolic changes co06.02
{w hours to 48 hours without food intake. ~-Saurce of blood alurase :
Non carbohydrate substrates are converted to glucose
Guconeogenesis).
Sourde of ATP for gluconeogenesis: The gets converted into
fatty acids + elyceral
Fatty acid undergoes beta. oxidation providing ATP.
‘@lyceral: Non carbohydrate substrate which can be
converted into glucose.
Prolonged fasting/starvation : metabolic changes : a-S days without food intake. ceereased
gluconeogenesis.
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‘Biochemistry + v4.0 + Manow 6.0 - 2022
‘Only source : Ta gets converted into FA + glycerol
TAG + FA + glycerol
dation feetyl Con Le. TCA cycle Ketone body synthesis Prolonged starvation : Metabolic
changes 00:10:52
more than 5 days without food intake, all TEA stores are depleted, fatty acids ore lou,
thetone body synthesis 9
muscle proteolysis > Cact ‘Finarrowedition 6notes mechanism of action of glucagon and
epinephrine + Glucagon is secreted when blood glucase level is FRR | FAD @ucose | FFA
major metabole fuel used : Free Satty acid > Glucose, He lyss.
Exceptions RBC and liver.
Grain camot use free fatty acid
A person had his dinner at € pm. Before having his breakfast 4s Y orTTnes eva gUtOse Wns
checked, it wos 100 ma/d hich pathway is the source of his blood glucose?
• eluconeogenesis.
• Hepatic glysogenclysis. “ masse chregerolse
‘Biochemistry «v4.0 Manow 6.0 2022
CONCEPT OF ENZYME REGULATION
Hormonal regulation, Allosteric regulation"
Hormonal regulation oo.o1:18
Well Fed/absorptive/post prondial state -> insulin High rstin agen rate Fasting sate >
ehucagon Lan iu gucagon rat
fetion of glucagon : ‘Glucagon binds to & protein coupled receptor. =
fetateDaengyeypiocenGnotes
cena a pecan oben HS erp
aTP—*> came
[Biochemistry «v4.0 + Mawow 6.0 2022
03 12 General
stony
Tnsulin
influence of insulin is active | influence
stote.
well fed state. Fasteg state. Enaymes active under the | Enzymes active under the
doe. dephosphorjoted, (inthe: phospho fated state:
of glucagon acte
fete state
[Slycolyss Dephosphoryjated state.
Substrate favour Sorward reaction. AA Feed forward
requiaton. Product hibit forward reaction. AeA Feed back hibition. anges ostere actvater_|
tasters evior m— Fructose posgrate [10 (Comert fructose sme Lp prose to Sucre vo curate
beghosgrate) apace ams me spies feetyconcarkouase | chvate efcon Gattg act spies)
Gourcachacery | Gatiyocds conceded fatty | sypivesaed apes) as coemayren dering)
‘Biochemistry v4.0 + Marrow 6.0 + 2022
INTRODUCTION TO ENZYMES
Introduction . o0ots0
Oefinition enzymes are specialised proteins that act as catalyst in biokogjeal reactions.
101850, Louis Pasteur discovered that sugar can be converted 40 alcohol This process is
known as fermentation. ve termed the ely vital hat catalyses this rection as Serments.
101897, edward Buchner discovered that cell free yeast extract could catalyse
fermentation.
Frederick w tiuhne coed the word enzyme which is derived. rom Greek word enzumos
meaning leavened.
Types of enzymes oes @marroweditionénotes
‘Simple enzyme : * Only amino acid residues tae part in the reaction.
complex enaume ©
• Amino acid residue + Chemical component i reagired for #he reacton to occur:

• Amino acid component & Known as apoenzyme.

• Non enayme component can be Inorganic. molecule lie metals : Fe*/Fe”, Cu, 2 > co-
factor Organic. compodient ike organic. molecule or metallo- organic molecule > Co-
enzume.

Trial aah 14 Enzymology 04

Properties of enzyme ones
eneymes are proteins. Exception : Ribozuyme | RNA ith catalyst property, examples of
ribozymes :
• 28 Ra in ribosome has enzymatic actity peptiy) transferase and is required for peptide
bond

• sn@an (spliceosome). ence

• Group I introns (oad.

• @nRse P : Post transcriptional modification of {RNA AL1CK ] iia trogen: a Precptoted

by poten precipitating agents.

4. Heat labile.
Coenzymes 1250
• Meat $88: rowedition6notes

• Low moleculr wieght non-protein organ compounds.

• considered as second substrates or cosubstrates. Examples most of the coenzymes ove

water sou vomins.
ete form Reacton vod tama 61 Thame + Gute decarbonjace | Puophosphate (TPF) | +
Transketdase tame 62 fof — (bose) + Redox reasons harm 63 (raced | nROv/ wre +
oerrogenases tame es on + Trancher of acy groups Geelfheris i) nee tame 6 Pupdocal
prosghate | + Trans reactions ie
i yoo |G) LTrancameaton
i a Transifuraton ham 69 Gok | Tetranukrofoh acd | + One carbon reactors
ochersisty + wi0 + Masow 8.0 2082

f T tama oa fdenosa + Tancher of (Cobalame/ meth 8a atom. afer cobamde enzyme a

Upoate + Oudatrve decarbaatin as part of mukenzyme | comple ke ppusate apracketo
gutarote | ;2etupsgmss, ranched chain keto acd dehyrogenose tama c fecorbate +
gdronjaton reactions Cofactor wae Metals as co-factor © metal
metal i Hight integrated by covalent or non-covalent bonds.
+o the apoenzyre. £4120 carbene arty EETOWedtionEnotes Cun tyrosinase.
metal activated enzyme : metal snot an integral part of the apoenzume, but ts presence s
reauired in the surroundings or the enzyme to act.
£4, Lipase requires presence of Ca” 40 act.
Prosthetic group Co 0258 Chemical component eoenzymes/cofactor) which is tightly
integrated to the apoenzyme by covalent bonds or non-covalent bonds.
Weloenzume fpoenzyme + Chemical component (coenzyme! cofactor).
iochemistry « v4.0 + Mamow 8.0 + 2022
@ dentify the enzyme involved. 1. Glucose ~-POA ———1—— > aiucose. Wao [3
Answer: Glucose - & — Phosphatase.
0 is added and inorganic Phosphate is removed.
An example of Hydrolase is Phosphatase.
Adding substrate 4o the name: Glucose - & ~ Phosphatase.
a. Glucose - & — PO, «> Fructose - b — POA.
Answer: Phosphohexose isomerase.
Glucose and Fructose are somers; hence enzyme is Isomerase. Both are hexose phosphates.
3.6lucose - 6 — PO, > Glucose
eo, (@marrowedition6notes
Answer: Phosphoglucomutase.
A type of isomerase: mutase.
The substrate is Glucose phosphate.
• NAD’ NADH. Pyruvate pest Con Yo Nw o
Answer: Puruvate dehydrogenase. In these reactions: NAD" = NADH: hence the enzyme is
dehydrogenase.
The substrate is Pyruvate.
Pyruvate Su 7 ovloacetate iotin Go.
! i 28 Enaymoloay 05
@ A person consuming Gaus ean develops upoglcema, Tiredness. Reason?
Probable Clinical questions.
Ansiuer Raw eqs contain Avidin, which Inhibits Biotin. Hence all the reactions catalyzed by
Biotin are inhibited.
example : Pyruvate , Oxdloacetate ao Go.
The enzyme Pyruvate carboxjase i inhibited. This reaction is a step of Gluconeogenesis.
Hence hen the blood glucose is reduced, Giycogen cannot be used as an energy source.
This results in Hypoglycemia and tiredness.
REE
Ansuver: Most drugs are acted upon by Cytochrome PASO and Cytochrome bs.
These re Hydroxylation reactions catalyzed by the Monooaygenase encyme.
@ Free radical scavenger.
fosuver: enzymes ore Peroxidase and Catalase.
In these reactions, HO, is converted to H 0.
Catalase is present in peroxisomes.
@ Free raacal generator. fnsuver: enzyme Oxidase. In Oxidation reactions
™, 0, Geduced)” x
od “Wo,
(ondized @09).
Bochemiy 140+ Marow 60-2022 MECHANISM OF ACTION OF ENZYMES
Active site ovore
The reaction i not occurring in the entire enzyme. it takes part only in certain site called
active sie.
Substrate
Retive site
enzyme
_Transiton state
fetivation energy
The difference between the free energy of substrate and he transition state.
The standard free energy change : As,
As,= Aq, - As,
ibs free energy tq
Biochemistry + v4.0 Marow 6.0 2022
TT hoyesoiveastn Aa, - Aa of ucatalysed reaction As - Aq of catalysed reaction ‘enzymes
lower the actotion energy. eraynes doo ater Ac, sandard free energy crorge. B15 es Sep >
erp
ERA WE dition6notes ©: enzyme, S : Substrate, P: Product.
Another way of reaction coordinate of catalysed
reaction 00:19:14. rca reacton cata easton 8 oe i eo i mp
Blochemisty v4.0 Marow 60 2022
[rages trngres entropy (dorderky of system. | Geduce the entropy. Sohaten hello hurogen |
oesovaton of the subst. bonded water around the
absiote
improper alent of reactive | Proper alment of rencte rugs enzgne tothe reaps he act ste fo
the
Substrate. Substrate.
Proper alignment of reactive groups in the active site to the. substrate Is explained by
various theories.
Various theories w2s30
I. emi Fischer's template theory/ Lock and key model: Fixed shape for the active site, so
that substrate. with complementary shape can bind to the active site of the enzyme.
a. witiam Jenk and Linus Bang near on 610 1es
fictive site of the enzyme complementary to the +ransition state 50 that more products are
formed.
3. Induced Ht theory by Dae Kosrlond (959). ‘enzymes undergo conformational chanae
when Substrate binds 10 4. weve sre, wnen induced. boy multiple weak interaction
between substrate and active ste.
Mechanisms by which the reaction takes place 003615
• Cato oy provi,
• Reid base calls.
• Coulent cata.
• metal on cata. §
• Cotas by strain. § I Catal by proximity
• For th reacton ta occur, he reactant shoud come ond forming distance with the active
site.
Biochemistry + 4.0 Marrow 8.0 - 2022 32 Engmoiogy 06
a. fed base catalysis ©
• Catolyfic residues in the acte site, either as a proton donor (Reds) or proton acceptor
(Bases).

• example :fspartate protease ~ they are he enzymes whose catalytic residue in the
active ste © aspartate (Pepsin's act ste ne ncarinte)

3. Catalysis by strain:
• pplicable for lytic reactions (catalust by Liases and. Hydrolases).
• Encyme bound to the substrate in such a ay that ‘strain and weaken the bonds to be
broken.
4. Covalent catalysis
• In Group transfer reactions. Examples : Transterases. Gransaminatior).
• Involves formation of covalent bond between enzyme. andi the substrate transiently,
• example Serine proteases (Chumotrypsin, trypsin, ogpatiroweditionénotes
5. metal ion catalysis
• Usually in case of metaloenzymes.
• example : zine in carbonic anhydrase.
• metal on s helping For the proper algrment of substrate +o the actwe sie.
hich of the folowing regarding lowering of actwation energy ore true except?
A. entropy lowered.
©. Desolation of the substrate. Confirmational change ot the active site during binding of
the substrate.
1D. fetive site of the enzyme and substrate are always complementary to each other.
Blochamisy - v4.0 Marow 60-2022 ENZYME KINETICS
Enzyme kinetics wo120
‘Deals with a things. aa .
Rate of reaction. Factors that affect the rate of enzyme catalijzed reaction.
Rate of reaction: on
Ave «5 peg ra
= Rote constant of |
famiel rene) | fowrdreacton ratPlial r= 6 ETE Ad mt carstant of backword reaction.
At equilerium, i & a dynamic state where forward and backward reaction takes place, at the
same rate.
er,
(aXe) =, (FXG.
FL = , Cauiorium constand.
BEX “va” Take)

,, = Concentration of products / Concentration of substrates.

• enzymes don't alter equilbrium constant (hed).
• enzymes lower activation energy.
• enzumes don't alter free energy state (AG
Ac? -2T log | 2 €ps constant. T+ losolute temperature).
‘Biochemistry - v4.0 - Marrow 8.0 2022 34 Enzymology
07
Factors that affect rate of enzyme
catlyzed reactions ore
• Substrate concentration.
• Temperature.

• W ions Concentration.

• Enzyme concentration.
Substrate Concentration : Increase in substrate concentration => increase in rate of
reaction > Ti # reaches v,__ Then no further increase in ate of reactions.
Te, nm Wo BTABHERBIER CS Substrate increases Rate of reaction also increases).
es Va x5] = 2er0 order Kinetics (ete sites are utiized No increase in rate of reaction). #, +
Michaels constant,
Leonor Michaelis & Maud Leonora Menten's
01507
1S) at Yo vmx = 20mmol = 1_ Find, _ V,__ = 40 mem. 15] = 1 velocity of reaction = X thenv__
= ax.
Significance of tm:
• Independent of enzyme concentration.
• Denotes attinty of an enzyme towards substrate.
iochamieny + wi + Marow 60+ 2002 + unique for an enzyme subsivate pair. £4, im value of
glucose hexokinase differs from K,_ ake of fructose hexoxinase. “Signature? of an enzyme
substrate par. * Higher the 1, Lower is the aSinity of the enzyme towards the substrate.
vm /a Soy Andy va (5) > wa (sl
_¥m+(s]
VX) 0 Vi max (s]
Single reciprocal plot a tade -vostee pot. ©) vanes-uoolf lot =
@ Catalytic constant (A Turnover number) ¢
• Measure of efficiency of a homogenous mixture of ene.
as fava
EEETTI—— 07 rome it 36 Enmymology. 07
ve. 8, (8, Total enzyme concentration).
ep.
To compare efficiency of enzyme : Calculated as teat / (en “Specificity constant.
‘Dissociation constant: ,
Rssociation constant : 1
Q Pancreatic ipase i an on enzyme that digest dietary Sats. fs an enzyme hat folow michaels
menten Kinetics. uo f rg Reg bot decries turncoat Feature of pancreatic Ipase?
velocity fhe: elf meximatishen 100 of substrate binds to ne enue.
8. Velocity of the s half maximal when 50 % of substrate binds to the enzyme.
C. Velocity of enzyme is maximal when SO% of enzyme bound with substrate.
0. Vekocity of enzyme is independent of substrate. concentration.
Q Expression of tissue specific isoenzyme is a method of regulation ofenzymes The glucose
metabolism differ in 26C ‘and Iver as one metabolse gucose and other store gucosedn 6C
rst step use Wexoknase | whereas ner gucoknase. what is the reason behind thi ?
i A. The rim of wn 1s higher than &K.
i 50s svensNoRaES 7mm an Gr The Kim of HA | & same as 61.
ovazss most optimum temperature in Wuman 35° to 40° C.
Blochemisiry - v4.0 Maow 6.0 2022 i -
maximum stabiity 45° 40 55°C.
Temperature Co-efficient @,) every 0° rise in temperature : Rate of reaction i doubled. qa
Rate of reaction is doubled because : * Increased inet energy. * Increased Collision
frequency.
eel shaped curve (Enzymes ore denatured
Optimum temperature Temperature
H* ion concentration 005507
marroweditionénotes
eel shaped curve
pH
Optimum ph Normal 5 40°. Changed state of fino acid residues aries with pH.
pr ston constant. i: “Charge of encable group varies’. fspartate protease > pepsin fete in
acide pH. Rspartate pi, =a. { Intestine = Chymotrupsin Catalytic residue : Histidine (ph, = 6,
active in alkaline pH.
‘Biochemistry 14.0 Marow 6.0 + 2022 38 Enamoloay
07
Enzyme concentration or0200
velocty proportional 4o velocity of reaction.
[5] vate)
Q enzyme with highest catalytic efSiciency (NICET march 3030)
1m of an enzyme = 10 micromole and. fica = 0 per sec. 8. 14m of an enzyme = 3000
ranomole and feat = 50 per sec.
C#m of an enzyme = & mieromole and feat = 300 per sec.
orm eye = and teat = 200 per sec. EEG RE =
@ In on enzyme cotalysed reaction substrate concentration was 1000 times Kimi of
substrate metabolised o 14 micromole of the substrate in @ min. If in the same reaction
mixture enzyme concentration reduced fo 1/3 rd and Substrate concentration is doubled,
How much te is needed for the same amount of substrate fo get converted +o product.
A9mn
8.135 min C8 min. 37min
Biochemistry + v4.0 - Marrow 6.0 2022 ENZYME INHIBITION
0024s
All cellular processes need enzymes. Control of these processes. an be done via. an enzyme
nhibHor. Types: Reversive inhibition : * competitve ichibiton. + uncompetitive hibition. +
miced iohibiton. reversible hibition : + suicide ichibiton. + Transition state analogues. Non-
competitive inhibition can be both reversible and. wreversivie.
Competitive inhibitior@Marrowedition6notes _sooszs
11s a. type of inhibition in which he ihibtor is a structural analogue of the substrate and
hence it competes wih the substrate For binding o the active ste.
eres sep E
ON
Properties of competitive inhibition:
• Inhibitor is a strusturol analogue of the substrate.

• Addition of excess substrate can replace the inhibitor Srom the active site.

Biochemistry v4.0 Marow 80 2022 hited

• 4 @arotigdifghenotes a xotercept + 7 + x-intercept moves towards zero.
V-intercept remain same. Plot is shaped like X.
Case scenario: 85 ear oid chid plagjing in the garden accidentally drank an unknown
solution Kept a bot. Later he was not feeling well. We toi his mother and she took him +o
the nearby hospital and. She olso took the leftover bottle. On the way he vomited tice and
wos experiencing abdominal cramps.
7 the casualty stomach lavage done.
i PR + S0/min and 8 : 80/50. muscle Fosceulations present in the legs.
Blochamtry v4.0 Manow 80-2022
¢ ERED me,
fins. Poison was Found to be malathion, an organophosphate. malathion
• malaxone

• Tohibits ich esterase (competitive ihioiton)

4 feety choline > ——— Acetate + choline i

fecumulates in nervous system + ‘Stimulates autonomic nervous system
i ‘Symptoms like vomiting, sweating, increased muscle tuitching
tially he hibition of Ach esterase by malaxone reversivle,
but 05 the dose of organophosphate increases, the hibition
becomes ireversicie. 3 (@marrowedition6notes
case scenario
1 Hooch tragedy, many people were hospitalised. Ethanol was.
ven as an antidote. why?
fos. methanol was gested in hooch tragedy.
methanol “I Coradenyde ——— Blindness.
Ethanol compeTIGeRy IISA te “ezine alcohol dehydrogenase and forms acetaldehyde which
i relatively less toxic.
excess ethanol replaces methandl from active binding site and prevents formation of toxic
formaldehyde. Non-competitive inhibition 001931
soother swe), SEE * £ © products Fp = po = [pa The inhibitor is rot strustural analogue of
the substrate.
Blochemisty + v4.0 + Maow 8.0 + 2022
[E—
The inhibitor binds to a distinct site. Erscos ep | biter (13 enaume-innbitor comple (6)
>enzume-nhior Substrate complex (619) > £+ © at a very neglgbe rate. Vmax reduces, 1,
remains constant,
he
' Ll
Lineweaver Burk lot of non-competitive inhibitor :
intercept = 17 Shape of V! o
Hinetic properties of non-competitive inhibition : Ves reduced I, (y intercept) increases.
Fam constant > (-D/#, Gc nkercept) unchanged.
examples :
ehioitor | emgrecnbted oon Cyocirome coudase Forde encase. lodoacetate luceraidenyde 3-
70, dehydrogenase
Sushendiy vids Mang 6h 208

0 SE ge Uncompetitive inhibition 0028.03

The hbitor has no a¥Siny towards free enzyme butt binds. 10 the enzyme-subsivate
complex
ers aere 2 [er Iohibitor enauyme-inhibtor complex (£) vr
Vedi | A Veuid
Vou!
q — adtharke sg
Lineueaver Burk lot of uncompetitne histor ©
| on or 6Brotes ]
x-intercept moves away from zero. J-ntercept increases. £9, Placental ALP ichited by
phenyllanine.
case scenario: Followng a short circus, a house was on fre at night. In the morring al he
Sarmiy members were Sound dead but none of the bodies were charred. What & the reason
behind the death of the oll the Famiy members folowing a fre? fos. Fire ~ Increased carbon
monoxide emission.
Blochemisy- v4.0 Marow 60-2022
1. Carbon monoxide inhists cytochrome ¢ oddase (complex V).
a. CO also has higher alinty for Oaggen
Mixed inhibition oss
@marrowedition6notes
9 | mmo. (od sper ns rrr in ~®
(Cd Ad (£3 >
UE
Slochemistry- wi0 + Mawow 602022
Suicide inhibition vas
The inhibitor 1s unreactive, i hijacks the mechanism of the enzuyme to convert self into a
more reactive compound. This compound binds 10 the acte ste of enzyme reversibly. Ao
known os mechanism based inhibition.
examples: + Alopurinol used For treatment of gout inhibits xanthine oxidase.
ante
oxase x0
pantie 5 xanthine —%— ure acd Wreversive hbo
© fopuredl 0, onartoe
• Rsprn ehibits cyoonygenase.
• oro methyl omithine inhibits ornithine decarboxjase. Ws used in the regiment; of
frupmAsHERstEs “Transition state analogue costa yy ——a > Torenste Product ~N
analogue ong)
Substrates gets converted nto a transiton state before forming the end product.
The active ste of the enzyme © complementary to this transiton state of the substrate.
‘rugs hove been developed which are structural analogues of
transition state of substrates.
£4 Penicillin s a structural analogue for the enzyme transpeptidase which i required for cell
wall synthesis of
bacteria.
lochamisty wi - Navow 8.0 3088
46 Enaymoiogy 08
meas.
QP antibiotic newly developed 1s a close structural analogue of a substrate that participate
in DNA synthesis of bacteria. Hence the antibiotic reduces the overall enzyme actwity but it
can be restored if excess of the substrate i added which one: of the Following best describes
the antibiotic?
8. 15 a suicide inhibitor.
©. fn reversible inhibitor.
€. competitive histor.
©. fn uncompetitive inhibitor:
Qmethand is converted to Formaidenude which highly tox.
Patients who have ingested methanal & treated with ethanol to
nbbit methanol oxidation. hich explains the rationale of this
treatment?
A. ethanol a structural analogue of methanal hence it can be a non-competitie inhibitor,
8. Ethandis a sucha) pnoiegug, of methand hence it can compete for the active site of
ADM.
C. ethanol can alter the v,__ of oxlation of oxidation of methanol by ROH.
©. Ethanol compete with the formaidenyde binding Site of the
enzyme.
mixed crease or decrease |v,_'+ v/a Gecreases)
Slochemisty + wi.0 + Mamow 8.0 2022 ENZYME REGULATION
In 19th century, Claude Bernard put forward the concept of
Control of enzyme synthesis wos #50 noun as induction and repression. £9 D Cholesterol
sunthess :
more cholesterd in diet
(@mdyrepressiéson notes Gene Sor Hm CoR reductase (rate Imiting enzyme)
2 vere synthesis Succi cof + skgene | ma spase san i vere. Increased heme ul repress ALA
suhase wharnas docseaced amount of heme will activate the enzyme.
Q Phensbarbitone induces heme synthesis and aggravates porpryia. uh?
Ans. Phendbarbitone gets metabolised with the help of uochromes, wih are heme
containing proteis.
hen cytochromes get used up, amount of heme decreases and turn ncices ALA synthase, Th
causes accumation of porphuyins (termediate), which aggravates porphyria.
Biochemistry + v4.0 - Marrow 60+ 2022 48 Enzymology.
09
Control of enzyme degradation + Short ved proteins eg, regulatory protens fie cupins. +
Roerrant/defective proteins. These proteins must be degraded which takes place in a
proteasomal machinery, These proteins bind to ubiquitin. + ‘Degraded by Ubiquiti-
proteasomal machinery,

ubiuitin-proteasomal machinery is a¥ected, the regulatory

proteins ie cugins are lost, causing dusrequlation of cell cycle which can lead to cancers.
Basic pothology of Azhemer's and Parkinsonism Aberrant proteins. ‘ ‘eSective ubiquitin-
proteasomal machinery, * @marroweditionéotes egpadation does. not occur 4
Accumulation of defective proteins > Disease.
Allosteric regulation
Allos : “other”. Rlosteric enzymes are those enzymes whose activity at Substrate binding
site/octive site modulated by the presence of a modier/efSector. substrate Uy este aoa sie
modter [> i A
Alosterie she
Positive modifer : Retotor (catalyfic activity creases). Negative modifier: Inhibitor (catalyfi
acti decreases),
Slochersiawy + w4.0- Mamow 80+ 2022
Alosteric action Positive modifer (m+) binds to on allosteric: site and. makes Favourable
conformational changes in the
catalyte sie, enabling binding of substrate to the actve site. Rlosteric hibition Negative
moder (1-) binds to onallosteric: Site ond mares unfavourable conformational changes in
the catalytic site, which inhibits binding of substrate to the active
seis] pe 9
m= whl [ro
Nas
Features of allosteric enzymes ( Correlate with hemoglobin) + most allosteric enzymes are
multi-subunit enzyme. + They possess a quaternary structure. + The modifer need not be a
structural anologue of the substrate. @marroweditionénotes + Substrate saturation curve a
sigmoid curve.
Soh,
• Follows a. sigmoid Kinetics as t exhibits cooperative.
binding, Binding of one substrate Favour binding of other
substrates to the same enzyme. ‘Does not Follow michaels-menten hyperbolic Kinetics.
(nich result n a hyperbolic curve).
Biochemistry v4.0 - Marrow 6.0 +2022
50 Ensymology
emmon ox actvaror > Graph moves fo lef, decreases > High afSaty or © state.
‘Addition of inhibitor > Graph moves 10 right, 1, increases Low a8Siniy or tout state.
Allosteric enzymes are of two types | series
increases. constant
v,., constant v,.. decreases,
a ERE 8 rrp ee
snare fetvator | ioniotor prove TT vere repartate warscarbomyase we ce (puradimine
supthes)
He of reductase Croesterst feet con corbosgase ovate Rey con Gatiy acd spthesd
carbon phosphate supthase-t Aeseety
Les spi) — _ |icasals Substrate Savours forward reaction.
Product inhibits foruuard reaction. Covalent modification 03238
‘Definition : Enzymes are regulated by addition or removal of Functional groups by making
or breaking covalent bonds.
Slocharsiy + v4.0 + Murer)
Types reversible: Partial protedlysis Ceynogen activation) + Teypsnagen > Trypsin, y
chemically cleaving polypeptide chain, * Chymatrypsinagen -> Chyratrypen. * In blood
coagulation, Rbrinogen — Fibrin. Reversible :
• Phosphorylation de-phosphor ylation (most versatile).
"i peekpaton
Smethepatin:
• OP ribosyjation.

• Ubiquitin addition mest common site for phosphorylation + OH group-containing,

amino acids Gerine, threonine, and tyrosine).

me
Provein ries “OS
. oR TrTaNSTEMENES %y se
foten peipatse
e-

stragen prosmnaryase ~ a
{ siyeagen degraded
‘ibeiiny*0teQtgeweh-ome
Enzymology
eetyjation Happens in ysy residues.
Enzyme is acety transferase.
Aeetylation happens in histone and other proteins.
Histone acetylated by Histone Aeetyate Transferase (HT)
Positive charges in histone reduced > interaction of histone with on decreases >
Euchromatin formation (less

sendensed form) This is an epigenetic modification.

OP ribosyjation +
‘Donor NADY
1. Cholera, toxin prevents @ protein from binding to adenyly,
cyslase. How?
Cholera, toxin enables AOP ribosyjation of & protein which
prevents it from binding 40 adleny! cyclase. @marrowedition6notes
2. Insulin and glucagon regulate the blood glucose depending,
on our dietary status. How?
Covolent modiReation,
Insulin Ged state) dephosphorylates the rote leiting enzyme.
The enzyme active under insulin is active in dephosphoryjated
state,
‘Glucagon phosphoryjates rate limiting enzyme.
enzyme active under glucagon i active in its phosphorylated,
state.
methylation + Donor of methy| group: $ adenosy) methionine.
Process of methylation epigenetic modification where cytosine residues are methylated >.
Decrease in gene expression
How does Yersinia pests cause plague?
Yersinia pestis induces production of enzyme called tyrosine phosphatase which disables
the phagocytic machinery oF macrophages.
fochaniatey "v4.0 tamew 00-2008
8S a Emme som Eis @ he subsate sous cure gen babu charaterces ‘an allosteric enzyme
system Which of the Following statement is true?
A. Allosteric modifer binds in a. concentration dependant marner# False, binding is not
based on concentration,
©. modifier can affect catalytic site by binding to allosteric site: True.
C Adding more substrate can displace the modifier + False, because they are not structural
analogues.
©. Allosteric moder changes binding constant not Vmax : Fase, as allosteric prune PAR
BAMBI RSETCS oF V series uhere KF V,__ can Vary respectively
Compartmentation 0080-22
Certain pathways take place exclusively in cytoplasm or mitochondria, or both. This
mechanism provides 0 sate space Sor certain reactions. £g, Fatty acid synthesis >
cytoplasm.
Fatty acid degradation > mitochondria.
eenesiens+eeeenee: 54 Enaymology
10 ET CLINICAL ENZYMOLOGY
Isoenzymes 00:18
Physically distinct Sorm of the same enzyme. Catal the same reaction.
Properties
• may be the product of diferent genes. eg." Salivary amyase and pancreatic amuose.

• may be made of different subunits.

4" Lactate dehydrogenase -> 4, Ham, Hama ete. Creatine Kinase > Ci~ me, Cri-mim, Ci- 66.
• Different electrophoretic mobilty. ©, Fastel 5 LorP4p RRL Ere: 5.

• Differ in heat stability, i e+ a forms of alkaline phosphatase > eat stable.

andBéReEBIEdilionBNOtes,
• Oifference in km or substrate specifiy. eg: Different afSinity for hexokinase enumes
from Iver and pancreas.

• Cofactor requirement varies. eg Cytoplasmic socitrate dehydrogenase needs NADP as

coenzyme ; mitochondrial socitrate deydrogenase needs NAD as co-enzyme.

• Different tsue ocolzation 7 LDH in heart, Lov- 3 in R8C.

enzymes Functional enzymes : Hage a Suncton in the blood. eg. Lipoprotein Ipase,
coagulation factors. Non functional enzyme No Function in blood. ‘Seen in Bod due to
normal wear and tear of organs, or injury to them.
aie Si a Rs 00 ees
used as diagnostic biomarkers. * Helps to find the location of the disease. + Reflects the
nature and severity of the disease. 2g, ‘A reversible inflammation in the cells increases cell
membrane permeability, and the cytoplasmic enzymes,
are seen in blood,
Jn iereversible cell injury and necrosis, mitochondrial membrane permeability i¢ also
increased, such that mitochondrial enzymes are olso seen in blood.
Lactate dehydrogenase
eupwate MOE deeregeA8 tt, Location > eyoplasm,
Structure > Tetramer with two types of subunits H and m. Based on the subunits, different
isoenzymes are seen,
soerayre | Sutuns | mokthy a Tbe 8 ata serum
pase tome | Fastest 20 tora fam [Faster fm ob tors fam, [Fast 20 toma fam, [stow © |
Lo & > Seen in severely il patients. Lox/ Loc > Seen in post-pubertal testes. @lood analysis
gives the total amount of LOH activity, To Know
the specific isoforms, electrophoresis s done.
‘Biochemistry + v4.0 + Manow 6.0 - 2022
e 10 gre 56 Enzymoiogy 10
Creatine kinase wie
Create ease
Creatine ME 0 onng phosphate. Location eytoplasm, Structure | Dimer with a subunits, M
and 6.
mit on168) | maximum eran
onaee | ntemedate | veart
oenzume | tectrophoretc | Tesue of orgn | Percentage bod " “ won
[(onsmm | east | saetamicce
CH mi: Creatine Kinase in mitochondria.
Allaline phosphatase (ALP) ME Removes inorganic phosphate buy adding water
(wdroases) in an akalne pi oenaumes
• 9% BHEioweditionnotes Located in epthetal cel of blary canalul.
marker of extra-hepatic cholestasis (marked elevation). eg, + Stone in common bilary duct,
carcinoma. head of pancreas obstructing the common bie duct. * @- a- heat labile ALP:
Located in hepatocytes. marker of parenchymal injury. Mid to moderate elevation. eg viral
hepatitis. * @- a- heat stable ALP: Placental form. most heat stable ALP. Inhibited by phenyl
alanine. « pre-Bap: Seen in osteoblasts. marker of bone formation. Elevated in Paget's
disease, vitamin D deficiency, osteomalacia, ° and 4° hyperparathyrodism. ALP is normal in
Multiple Myeloma.
Slochemiery + v4.0 Marrow 6.0 +2022
10 Gina Enzymof © camma mp: Seen in mucosa of intestinal cells. ‘elevated in Uicerate
colts.
Cleared by sinusoidal cels of Iver. In case of hepatits, it wil be elevated.
• Leucocute rp: marker of leucocyte disorders Ike leukemia. 50 called carcino-placental
enzyme. Siar to placental ALP. erm cell origin. ‘Depressed in conditions of malignant
tumors.
IE capi seen met recs
Cardiac Biomarkers 003038
Rise when there is injury to heart.
enzymes and. pres and none Same we dition Brotes
Name. cise Peak | Return to base ine Soe [Evan we] wen oie [Fata [vim] eos Fugit | toad |,
we | Fas EL [eva | [Cor [ran [vas [ems or ander are abs
cardiac troponins are more specif than Ch- me. First biomarker to rise > Myoglobin (a
hour) => Kon specific.
igh sensitivity monoclonal ib based assay
Detects <1 ng/L of troponin. i Flipped LOH pattern
In normal individual > LDH-3 7 LDH- I INI > LOH-1 ? LOH-2.
Slochemisty + v4.0 Mamow 8.0 2022
58 Eomoony 10 - rain notriuretic peptide (BF) Not a marker of schema. marker of volume
overload and circulatory Failure. Enzyme profile in liver disease wane
© pet.
oy
Slochamishy + ¢« —_— 10 Sor
From the previous table.
ALT & more specific for Iver injury.
(very high values only Sor Iver injury)
FST verge or rar, er ltt sc,
Kidney and pancreatic injuries- non-specific. 7 erty reat FST ate fon toh AST + ALT ratio
(AAR) ca we,
• Chronic vol hepsi
• Non-aleoholic fatty liver disease.
• Toe hepatite.
• scetamingphen toxeity were J — Damage to mtochondria results in rise of ST eves »
opadc enous: ST levels creas fe sadn col 8 er are injured, and AST i not cleared from
the plasma.
• Lier reopiasia aminotransferase level — f- pos La rat . eins u —
feute injury or inflammation of Iver = Marked elevation of te Chronic conditions > mid
elevation of transaminases.
A er enzymes within normal lis in an asymptomati son, ith only mid elevadion of ALT > Fay
er:
Biochemistry + v4.0 Marow 60-2022 — 60 Ensymoloay
10
fale phosphatase (LE) fresent in the cell membrane of epithelol cels of bilary canalicul
aoe
marked elevation signifies extra- hepatic obstruction. Non specific marker.
<' nucleotidase + Specific marker of cholestasis. Present in cell membrane of bikary
canalicular cells.
Gomera, gta transpeptidase (60): Located cel membrane and smooth endofiasmic
reticulum (merosomes) of the hepatobiliary system
Less specific for cholestasis.
Glevated i ntra-hepatic and extra hepatic chokstass. marker of marrowedition6notes
loohole Nepatite. | ovement of microsomes. voy drovers, |”
‘rugs like phenyton and phenobarbitone.
art (=
apt = crest en cuckeotdase 1 1 Enzyme profile in prostate ouss1s
reid phosphatase + Inhibited by tartrate ion > Tortrate bie. (That is found in spleen
leucocytes ~ Tartrate resistant.
Slochomisy + wi0 + Mavew 60+ 2088
>_> 10 re ome Not a specific marker (ako elevated in osteoclastic iguried. Prostate specie
antigen + Aco called Kallikrein related peptidase-3 (aun~ 2. Cut off > cAng/ml
00:56:55
Non specie as itis elevated in other intra-abdominal conditions, salivary gland déorders,
genitourinary lisorders lie ectopic pregnancy,
Lipase +
elevated almost $000 times.
marnerso bones one formation one resorpten
Orgates from osteoblasts. | Orgnates fam ostecclasts.
• Pre- beta we oN co
[sora OTHERS DEN Sos
+ Pro peptide of type reotagen.|* urine free deoupyrdecine
Novel markers of acute kidney injury: + Kidney injury molecule (aim D- * Neutrophil
gelatin associated ipocalin (eA). ore + fr. * Glutathione S- transferase. + sar. + Geta. a
merogbbuln + pha. a macroglobulin + 2etino binding protein. + cxtaine ' + microaloumin. i
Osteopontin. + Liver Fatty acid binding protein. * Sodium hydrogen exchange isoform. *
exosomal Fetuin.
Biochemisty + v4.0 + Marrow 6.0 + 2022 62 Enzymonogy
10
Marker enzyme of cell organelle 01010
Plasma membrane i q rucleotidase. Adenyy cyclase. ; NC ATPase. endoplasmic reticulum >
Gucose-6-phosphatase. Golgi apparatus > Galactosy transferase. mitochondria, ATP
suphase. Lugsosomes = Cathepsin. feid phosphatase.
Clinical scenarios oro120
QA 50 year od man presented with complaints of squeezing type of chest painuhie he
climbed up the stairs. on examination, in anus and restiss, tachycardia present, 67
50/iomm 4 Hs 28 soy of DH evaton what & the rect investigation to be done?
Suspicion of mi. Troponins elevates earlier.
QA Toyear oid male patient was admitted wih generalized weakness, easy fatiquabity,
#ehing and jaundice For one month. On examination, patient was emaciated and seratch
marks were seen all over the body, He was jaundiced and a mass was palpable in he upper
abdomen. which i the best ‘enzyme marker of this patient?
A Rlonine aminotransherase.
6. Akalioe phosphatase.
9 rucleotidose.
©. Aspartate aminotransterose.
Biochemistry - 4.0 Marrow 6.0 2022 est marker ts §' nucleotidase.
Qf leyear oid female patient was admitted in medial ward with fever, anorexia, vomiting,
generalized weakness and jaundice. She gives history of recent travel to Bangalore. Acute
rol hepatitis was diagnosed. which is the non- functional enzyme elevated in this case?
A Rkaline phosphatase.
© Alanine aminotransferase.
©. Gamma. gutamy transferase.
©. Creatine kinase.
© Lactate dehydrogenase.
QA 38ear od lady wos admitted Sollowng Road Traffic Aecident. During her hospital stay an
intern in the ward whie “apmEgatswpy en. cass Rie noticed that her serum ALP wos
‘levated and Skull X-ray showed-Osteopoross cireumscripta, Hhickening of diploic areas)
pad rsslerasisi of portions. of skull ones. Out oF his curiosity he sends a. blood sample to
Biochemistry ab. which of the following s the Biomarker he sent?
A Serum osteocalcin
©. ProBnp:
C. Ane
0. imi.
& Bxosomal Fetuin-A
Q. 40year od female with history of inadvertent use oF NSAIDS develop oluria. Her Serum
creatinine and UN ‘eevated and Route ney Injury Can] was dagrosed. which of the folowing
are novel markers of Al which can be done in this case?
AN ferminal pro exe.
©. Custer.
Neutrophil gelatinase fssociated Lipocalin
©. Cross Inked n-Telopeptide.
Biochemistry + wi.0 + Marrow 8.0 2022
10 Bet 64 Carbohydrates
1
CHEMISTRY OF CARBOHYDRATES
Definition onu Hydrates of carbon. ceneral formula: C, 4.0),
where n the no. of carbon atoms.
They are ldehyde/keto derivates of poor ako.
Classification of carbohydrates 06:15
monosaccharide : Sngle sugar unit ‘Disaccharide : Two sugar unis. Shpsrants omer oi
Polysaccharide
units. @ fo AT
00018,
Blochemisiry - v4.0 - Marrow 8.0 + 2022
0548
Depending on Which carbon HIE BreRRMEER Tr ‘gueosidie bond formation, they are
divided into reducing and nonreducing disaccharides.
Non reducing | Functional groups are engaged in linkage, and 0 Sree group present.
Reducing : Free functional group present.
examples of reducing disaccharides vame | monomer yrs maose | clucose + Gucose | Gi 4
gucose inwage somattose | Glucose + lucose | ai - & gos lnkage Loctose | Galactose +
Glucose | fi - 4 gucoseic Inkage Lactose | saloctose + Fructose
examples of non-reducing disaccharides
Name monomer units Linkage sucrose | lucose + Fructose | a - Ba lrvage Trehokose |
Glucose + Glucose | au -ilnksage.
Polysaccharides oo coon Homopolysaccharide sila
one type of monomer > pe of monemer us. Glycogen
• Storage carbohydrate in humans/onmals.

• made of a gucose.

• Branched polymer.

Blochamisiry + 4.0 Marow 8.0 - 2022

Straight chain OL 4 linkage.
• A branches: 0.1/6 linkage.

• Starting point has a reducing end,

‘muttiple non reducing ends present where glucose. molecules are released present.
Also called non starch polysaccharide.
Definition : Remnants of edible part of plants and analogous carbohydrates that are
resistant to digestion and absorption by human small intestine but completely or partially
fermented by human large intestine.
vuran Hees areas oho he Bi-+ linkage. most oF cellulose is made of BD ghucose Usually
Fermented to short chain fatty acids live acetic acid, ‘butyric acid 4 propionic aria (energy
source).
dietary fores L y Insoluble (erude Sores) + souble: Celulose Pectin Hemicelluose came
Lectin muciage
Lectin connot be digested, absorbed or fermented.
208 + 40 9/2000 Heal. energy released per gram: a Hcol/g of dietary fibre.
Uses: * Ret 0s substrates promoting colonisation of probiotic strains thence are prebiotics).
* Faecal bulking and softening, * Regularity to bowel movement. + Reduce cholesterol by
binding to bile solts.
[Biochemistry « v4.0 « Marrow 6.0 = 2022 1 Ei + Favours satiety,
• Fibre parkiularky aus Genuareek) and pect reduce
postprandial ood glucose level in the ood.
ulin
Fructose, Fructosan.
uli clearance fest Used to access, glomerular Mration
test (deaD but not used now. Chitin In exoskeleton of crustaceans. made of N-acetyl D
glucosamine.
ecto eterapaacchurde. Predominantly made of galacturonic acid,
‘Dextrin Hydrolytic product of starch. ‘Dextran: Polysaccharide of GD glucose.
• used oa plasma. oems expander

• Supthetc docian Se eckson dvomatogaghy

• antl ou. Tsomeriom in carbohydigtes,.. eitonsnotos— 234 Isomers are compounds

with same molecular formula. but different structural organisation.

4 Structural somerim/ optical somerim Stereaisomerism

DL isomerism (enantomers) Destro or evo rotatory
i naan NC —— 4 alencies of carbon atom occupied by 4 diferent oroups. ce “7 I I! c bw 4S y i
wl, 1/4 x i de Kp EE Ww Cm i
lochemiery «vi0 Marnow 80+ 3082 11
Bet fg 4 asymmetric C atoms.
0038s
Tom prion of dicen vue atten corbon tom changes at the penultimate/ reference carbon
atom. They are mirror images.
most carbohydrates exist in © form. most amino acids exist in L form,
in glucose, depends on positon of 1 ant O on °C atom. i
a Lon
L{
n @marroweditionénotes om O-Giperadenyle L-siperaklehyde D-gucose L-gucose
Anomerism 04925
Isomerism at the functional carbon atom. examples + a glucose and B glucose. wo
cron bie enon Wi T ATTN oe WAT Ae on wh I, = owl +o tn wo i Wooo wen ot ccose | nto
Batusose wc —ou | on woe con ha] dq Ni w, "fou MHjem ww oH ow a-o-frutofuranose B-o-
tructoturancse
Cham ST ate Epimerism
Oifference in orientation of and oH
roup at a. singe carbon atom other, {© a
Han the Sunch peru i i te inctonal ond imate WC oun
carbon atom.
Dasteresisomers Difference in orientation of H and OW groups in > | carbon
‘atom other than the functional and atom.
Optical isomerism srr
itty to rotate the plane polarised ight. Occurs due to asymmetric carbon atom. * Glucose:
Right/clockuse direction: Dextrorotation
Cd glucose or +). Hence glucose © known as dextrose. * Fructose : LeM/anticlockusise
direction : Levorotation
4 gusose or). Sucrose tally dextrorototony and after i is hydrolysed, becomes evogatatoru.
Hence aka nverk sugar.
¥
a at ! |
a
Biochemisky + wi0 + Maow 8.0 +2022
70 Crvohygates 11
One liners
[Ee
Carbohydrates uh no asymmetric carbon atom :
‘Dihydroxy acetone. Amino acid with no asymmetric carbon atom © Gycine.
most predominant Sorm of glucose : B D glucopuranose. most carbohydrate exist in ©
form.
most amino acids exist nL form.
(@marrowedition6notes
Biochemistry v4.0 Marow 60+ 2022 GLYGOSAMINOGYCANS AND
MUCOPOLYSACCHARIDOSES
Introduction 00008
Sores”
Gepeating deaccharide unit so called as Weteropolysaccharide/ mucopoksaccharide
Long unbranched heteropolysaccharide consisting of a. repeating disaccharide unit
Properties of epee Negatively charged/Poljarions. + Form hydrogen bond with water. +
fbsorb water and form a hydrated gel acting as a ubricant. L Compressivitty of cartiage. a.
mobility and resiience of joints. + Occupy lorge space —» Molecular sene. + Selective
transport of molecules.
Bochemisy + W0- Marow 80-2022
12 72 Carbohydrates
1238
Chondroitin Sulfate © * Compressibity of cartiage.
Weratn Sulfate © most heterogenous 6AG. + Repeating unt is Galactose and hence no ron
acid + #51: Comea (Comeal transparency). + KSI: Loose cometive tissue.
Dermatan sulfate +
© widely distributed in the skin.
• structure of sclera.

• Atherogenic a : Synthesized from arterial smooth muscle cells, hence attaches to low
density Ipoprotens (DLS, These LoL are carriers of bad cholesterol

Blochamatry v4.0 Manow 60-2022 oo

Heparan Sulfate :
• Present in synaptic vesicles.
*+ Present in the renal basement membrane making it responsible for the charge
selectiveness of the glomerukWeasenientmnenensfe:
• Gives a net negative charge to the basement membrane, therefore doesn't let
negatively charged, Albumin to pass through,

• Anchors lipoprotein Lipase to the endothelial surface.

• ets as plasma membrane receptor.

Heparin +
+The only infracelular 6A6,
• In} Heparin dslodges lipoprotein lipase from its anchoring site.

• Naturally occurring anticoagulant which binds to Anti-Thrombin I

Hyaluronic Reid, Not covolenty bend AP: AMEBtion 6notes * Polysaccharide.

• Help in cet migration ound epat, Embryogenesis, morphogenesis, metastasis).
Proteoglycan Structure ante
Stak
rrtencre i
Biochemistry « v4.0 = Mamow 6.0 - 2022 74 Cosvonyarates
Ge(35#) + Proten(m = Proteoglycan. Bottle brush shape.
Mucopolysaccharidoses (MPS) 02554
es ore supheszed in the endoplasmic Reticulum ond ei) fpporatus.
They are degraded in the Lusosomes.
MPS ave a group of disorders affecting the degradation
of 6a in the Ligsosames, hence leading 10 intra. lysosomal accumuiagifharroweditionénotes
‘They are essentially, Lysosomal Storage Disorders. nneriance is Autosomal Recessive
(Except Hunter's Disease) . Mos commen mS + Sanfippo? Hurler > unter .
‘General features of MPS * Coarse facial features. + Frontal bossing + Depressed nasal
bridge. + Corneal couding + angualhuperoghy + macrogossia leading to recurrent URTIS
and hearing loss. + Clwvond, 3 + intellectual dabilty, P ~cemvevmseRRbeguly leading to
umbiial o a inguinal [I
• Short stature. Radiological Sndings *
• Dysostosis multiplex.
Biocnamatry v4.0 Marow 60 2622 + eaking of vertebrae.
• Tip of metacarpal degeneration leading to
Bullet shaped phalanx.
Hurler's Disease/ MPS, 1-H * mutation of IU Gene.
• Affecting a-L idurondase enue. Seheieks Disease / MPS 1-6 ¢
• mutation of 10UR Gene.
• ¥ecting @-L Iduronase enzyme but the enzyme activity i partially preserved.
• Normal inteligence.
• Presenting age & abter 5 years. vnter Disease/ MPS 1;
• Mutation of 105 Gene.
• Affecting duronate Sulfatase enzyme.
• X-Linked Recessive condition Henpecinky moles are
• affected «Clear vision. (ub corneal couding) Notowiez Syndrome! mPS Ix : AtSecting
Hypluronidase enzyme. 5 (= wap | | oefcency | Cloudng mes 1 Grier) | duronose
present | present res vs Crm) | roe Tsers [preset PSN Gumte) | kuonate Sufotase |
resent | osent 05 1 Gonfigpe) | enzyme that degrades | Present |ssent | — {aparan
 Suftoty. - 06 (ror) | soloctosomine- Gosert | present -suftase, seta-calactosdase wesw
re Present (reroteausc Lame) | -sufotose I
Blochamisy v4.0. Marow 60+ 2022
i 76 Carbonyarates
12
Osease | veceromeaply | short | oxgontone [Laure
Stature |multpex | inckson | ey eos nchir)
MPSin [Present |eresert resent | preseot |
Gurieo)
mPS[Present | present | present | present
(shew)
[mesu Present | Present | present | present
Gunter) |
[mes Present | present [resent | present
Gantippo-
fe mes
mes sent Present [present | osent |
(roca) |
wesw Present | present | Present | present
[—
pe) |
(@marrowedition6notes Treatment Modalities 004303
Stem cell Therapy : MPS 1-H and MPS 1-5. mPs vi.
enzyme Replacement Therapy:
mps 1:
Adurazyme.
MPS 11+ elaprase. + Noglazyme.
mps vi
Stil under trial
Substrate Reduction Therapy * MPS IN Flavinoids.
Bocnamiry 40 Marow 60 +2022 Clinical scenario 1:
6 year od mentally retarded ari uth protuberant abdomen, Umbiical vermia, short stature
Answer : Mucopolysaccharidoses.
Explanation : mental Retardation intelectual diab Umbiical Herma.
Short stature.
Corneal Couding,
Coarse Facil features Frontal bossing, depressed nasal bridge, macrogossia.
Claws bond.
81 0F which are Features of MPs. SE &matioweditionénotes
Clinical scenario a
fle year oid boy usth short stature, coarse facial feature, and hirsutism ahd normal
intelligence
fosuer : Schee's Dsease Explanation: 1 year od boy with Short Stature, Wrsukm, Normal
intelligence, Coarse Facial features,
Cormeal clouding Aodominal protrusion
Claws hand 1 L-lduronidase enzyme s fected. we mn conclude t to be Scheie's Disease.
Biochemistry + v4.0 Marow 60-2022
12 Giycosamino
gvcans. Mucopol
i
Clieal scenario 3 Qa year od male presented with coarse facial features, intellectual
disablity, prominent abdomen and dear Vision, What i the diagnose?
Answer + Hunter Syndrome (x-liked Recessive)
‘explanation : its a. male chid with clear vison which ndicates. Hunter syndrome os it
exclusive spares cornea. and is a X-linked recessive disease.
Clrical scenario 4 6 year od mentally retarded gel ith profuberant abdomen, umbiical
hernia, shork stature along with Solowing Siding seen.
be shaped middle ea ody inclusion pala
Answer : Not MPS IV
explanation Leukocyte inclusion bodes called Reilly bodies are seen. eullet shaped middle
phalanx also seen. These features are exclusively absent n MPS IV. GLUCOSE
TRANSPORTERS
Types of Glucose transporters 00154
‘Soduim dependent Glucose transporters : SGLT. Sodurm independent Glucose transporters
| GLUT,
hucose a huydrophiic compound, t must be transported. Hough the hydrophobic plasma
membrane. Hence Transporter is required.
SGLT 00:04:28.
Tope [amt Fancher
Sau | kesboe Lomi sd. | sorption of goss. | roseral Renat ines.
S74 | Provemal end bles owe SECTRERD gh
SeiT-1: Characteristics
• Sodium - dependent.
• unidrectional transport.

• Secondary active transport.

• 15 asymport : Transports Glucose along ith Sodum.

Na ATPase pumps 3 Na” out and a 1 in wih the tization of IRATE. Hence in SLT, Na’ i
transported along the concentration radient while Gucose transported against the
concentration aradient,
eran SUA pent recuire RTP but 4o pump out the Na and 4o bring in K (to maintain
neutrality) the Ne/k ATPase pump utizes ATP. (secondary actwe transpord
Biochemistry 4.0 Marow 8.0 - 2022
13 80 Carbonate
Lumen Inkestralcel Good & awe i Jo ax
Clinical appications.
• In ORS mmr msersesave supplemented together os they are absorbed together.
a me oS BaP YB ttn oh is gene ree om St cae
Hence the reabsorption of glucose i inhibited.
The renal threshold is lowered (normal 1Bomg/aD.
Hip ire tal iingiod concentration of glucose. i es ld Sora vi i are wh ol pennies called
&fiflozins.
ar poses li
Side effect : urinary tract infection.
GLUT wuz
Characteristics
emse nny
• Sodium independent.
• Faciitated carrier-mediated transport.
• Passive process.
Biochemistry - v4.0 Marrow 6.0 + 2022 a oncom 1 sor gmat * Bidirectional + fog the
ecertaton gant * Ping Pong mechanism +
eee pecans —Tle/} = ance 0if j}———}*|_- Png sate Pong state vo osentin
Png state — Faces towards high concentration of glucose and attaches to it.
ong state — Faces towards the interior of the plasma membrane. Ping state
conformationally changes to Pong state and glucose 1s transported along the concentration
gradient.
• Blhyperbolc curve cenypaeiyMGUEM EMS tos
concentration graph.
[> Serpe aiusion y 0 (5)
‘5 — Solute concentration V— Rate of absorption of solute,
Hyperbole curve - The rate nf absorption of solute raises i initioly as the solute
Concentration inereases and remains i the same after a particular concentration (vmasd.
Reason ~ all the carriers of glucose get saturated at V max
In simple diffusion, 5 is directly proportional to v:
[Biochemistry « wi.0 « Marrow 6.0 + 2022 2 Carbonydrates
13
Sur [locaton Concept/ Features wr [ean Codey devia
placenta, dney, vigh a¥Sivty Flow K,
6¢, colon retina. Basal glucose uptare.
Seen with Cell barrier mechanism.
&uT-a |B cells of Pancreas. Low afSiny, K,_~
Sinusoidal cells of Liver. | high.
Basolateral/ serosal | Insulin secretion.
side of Intestine. Uptake gucose Sor
Basolateral side of | Storage.
Procma renal tubules. | Absorption of Glucose. Reabsorption of ahicose.
SLUT-3 | Neurors, news, Vighest afinty to placenta. gueose > low: SLUT-4 (Dear adposie
hele} Inuin-dependent. muscles vecrease bod, ucose (maximum extraction - of gucose in he
postprandial state) SLUTS | Luminal se of Fructose transporter. the small intestine,
spermatozoa.
SUT-G | Spleen, eucocyfes. | Pseudogene. aur
LUT-6 + seen in Blastocysts. SLUT-9 : rate transporter. Mutation oF 6LUT-9 & a cause
of Gout.
Biochemistry + 4.0 - Mamow 8.0 + 2022
003553
Luminal side of Intestine ©
Absorption via. SeLT-1 + Galactose > Glucose > Fructose. SLUT=S : Carrier mediated
transport of Fructose along the concentration gradient. t i Faciitated diffusion.
%
eur -a/ Gyogen
SLUT-3 + Transport OF Glicose, Galactose and Fructose from intestinal cells 4o the blood. It
is Insuln Independent.
10 cells of the Pancreas and Liver, Glucose & transported. into the cells via LUT-a.
has a low a8 Sinity to Glucose.
Hence i is transported when the concentration of Ghucose is more in the Blood.
In the Iver, Glucose is stored as Glycogen.
In Pancreas it helps in he secretion of Insulin.
In the Presence of Insulin, GLUT-4 is recruited 4o the plasma
Blochemisry - v4.0 - Marrow 6.0 2022
Gicose, 13; Jucoes oem -_T membrane of Sneleto scl, dose Kssue, and heart The excess
gucose the Hoods reduced cose not he ory source of energy hese organs.
Distribution of glucose transporters in various organs oa
Brain GLUT- 1§ GLUT 3 both have a high afi to ‘gucose, hence glucose i transported even if
blood gucose levels are very lows.
RBCs: GLUT
Placenta: GLUT GLUT-3. uth hgh fetal glucose demand, ‘hucose i transported to the Fetus
even i the blood glucose levels in maternal ood are fou.
Neuronal Glucose Transporter : GLUT-3. widely distributed Glucose Transporter : GLUT-L.
hucose hs high siceme index than Fructose. Reason : GRIESE ABLOAIRH YAHEST-, against
the concentration gradient, whe Fructose via GLUT-S which & Sacitated diffusion.
Glucose, Maltose, Lactose, and Galactose have a. Glycemic. index of 1 or 100%.
Dietary Reres have a Glpeemic index of 0.
Question : A S-year-od boy aber drinking contaminated water resulted in acute watery
drrhea, later diagnosed fo be cholera. + what i the role of Giycoipds here? ‘ami Gangjoside
acts as a receptor for cholera tox. + what's the role of 02S although electrolytes are lost
stools? Na’ and Gucose ve absorbed together via SLT) Gharib a et mio electrolytes in took. +
1 we add amino acids, wil # aggravate the situation? No, Because the transporter for amino
acids 5 different.
‘Biochemistry - 14.0 Marow 8.0 + 2022 14 5
GLYCOLYSIS
Concept = 00217
Podhuuay taking place, when body is n a wel-fed state (igh insulin-glucagon ratio).
aucose — gems ios i out HIP pata ape HI |
Pate feetycon so UE ocve |, N
TCA cycle.
arroweditionénotes Fatty acid
1 «Tracy gycerol
Glycolysis : Overview w0s40 Ao known as Embden Meyerhof Parnas pathuay (emP
Pathos). Derived from ereeks word ‘aus Sugar. Lupe! spitting. Sie: All organs. organelle +
Ctoplasm. significance + i + Only potty that takes place aerdbically and anaerobically. + 80:
Onkt metabolic fuel in nolaadasstbaskaarstkg s
entrely glucose.
‘Biochemistry + wi0 + Marrow 6.0 + 2022 I -_r
Ori anaerobic geass occurs in B6C as RBC lack mitochondria. Defect in gyeolyic enzymes
REC lysis.
Soonaus capacity for gisokge.
exercising muscle + Decreased oxygen supply ond hence
derive supply from anerobic glycolysis.
Defect in gyeolyss : Musee fatigue.
veart:
Low glycolytic capacity : Cannot survive ischemia.
Steps of glycolysis
Preparatory phase + Stages of phosphorjatien. + Stage of spitting, This phase utizes ATP
i a. Pou oF phase:
i + Stage of oxidative phosphoryiation. This phase generates ATP.
Blochamisy 40 Marow 60 2622 Prope pre
(Suoose ) HXHERSE, (cme proghote ) we oop
vem | ogee
arp — 1hope
rere
oe ord
orgirosy acetone | | serie >
pre 00) | |r Gy >p) rocqrorie corinne
(me) Mme ie)
[Led
°
Signifeance : + Pirreversivie step. + Requiatory step. * Fl generating step. + Phosphoryjation
by hexokinase traps ghicose for celular metabolism. 2) in somerisation of el phosphate to
fructose ( phosphate, the C=0 group is transferred to Ca 0 that a free hydroxy
ofoup is avotable ot Ci 6 areverside step. » (Fria phase)
1970 oss ctonnase
Fructose vo biprosphate
Blochamisy 40 Marow 60-2022 8 Carbohydrates
14
significance : * Rate lmitng step + aireversile step. * Committed step of glyeokyss as
fructose |hisphosphate Formed can enter only glycolytic paths. * eotile neck of the
pothuuay. Fructose yo bisphosphate & spit into two 3 carbon compounds, dinydroay
acetone phosphate (oF) and ghyceroldenyde 3 phosphate by the enzyme, aldolase. This s a
reversible step. OHAP gets converted to gyceraklehyde 3 phosphate by phosphotriose
ismerase. 2 molecules of glyceralsehyyde 3 phosphate enters the pay of phase.
vouasinase [ Glucoknase + Has 4 isoforms. + exokinase IV. + wighofSiity, ‘Lowery |
Lo %@marrowedition 6188" ur Not an inducible enzyme. (+ Induced by insuln. +
Vousekeeping enzyme. | * Induce enzyme. nbibited by 6P0, + Not inhibited by 66PO, ~ +
Regiotes bod glucose in post prandial state. * Present inbetacelsof | pancreas § Iver.
Phosphoryation by hexokinase fraps the glucose for cellular metabolism.
Ghucose enters the cell via GLUT 4 transporter which allows. bidirectional transport of
gucose.
However, bu phospharviating ducose, ts transport outside the cell i prevented and glucose
phosphate now enters. varius. metabolic pathways. Phosphofructokinase | (PF D an
allosteric enzyme. Allosteric activator of PPK 1s fructose ae bisphosphate. Fructose 6
bisphosphate is formed from fructose phosphate by action of the enzyme PF a.
Blochamstry 40 Marow 60-2022 14 on 8
PAH His inducible enzyme.
Pay off phase __ ows7.06
ax| syperalargte (3 posphate
y3 Fo, derysragenase eae ceversbe step vrergone Po, added
NOt ago = as ATP x a >see
POH EH:
teguctory top 2 ane x a= ane Paywate ) Anaerobic glycolysis ooe19 ducose 4
Biochemistry: v4.0 = Manow 6.0» 2022 90 Carbonyarates
st
14
No net generation of ADH.
Anaerobic gyeoiysis takes place either wien there & no mitochondria or when there lack of
oxen.
Significance of anaerobic guyeolysis n Rec
There is no mitochondria in oF NRD* wil not occur.
mature £6c, hence regeneration
Lactate dehydrogenase (0H) in onaerdbic glyeoiyse.
regenerates NADY.
This also prevents accumulation of NAOH (product) and. ensures that glycolysis is not
hibited.
Energetics wsoss 0 hero goss ©
fers eidog steps | umber of TP generated lperadenugie 3 NRO x3 + & NAOH - 85 x3 =
prosghate sare i
2 ope KRUBBITOWETTON PE = orp
Pupusate Hnase Imex a ane
50 net ATP Generated 19 - 2 14TR.
aerobic goss
energy yeng steps [Number of TP generated | 12606 ase [rere x aca me
From | glucose bu aerobic oxidation : 9
Bochamsty v4.0
Maou 60-2022
Aerobic glycolysis = 1ATR POM=asxa=SATP
TCA cycle = a x10 = 30 ATP Total ATP generated : 33 ATP.
Inhibitors of glycolysi
• Arsenate : Decreases the avalabilty of Pu
a. enolase Inhibited by Suoride by reducing the avalabity of mg or me, Clinical
significance {NaF oxalate mixture used in estimation of blood glucose.
Rapaport Leubering cycle
Taxes pace inside Rec. Only 0% glucose enter thigkgeierowe dition Boles Ao called RL
Shunt / 3, 3 606 Shunt.
Biochemistry v4.0 Marow 60 2022
92 Carronyertes 4.4 There is production of 4,3 8P@ by shunting the ATP {generating step
in glyeolysis. aera:
• Shit oxygen dissociation curve to right (ow af Sinity state).
• Helps in unloading of oxygen from Hb. No net genecationa® arm, 10M oF glucose in
28¢s enter this pathway, Regulation of glycol ores |. Hormona! regulation 2y covalent
modieation
• Fedstate: pemin iabngs (me @marroweditipognotes : cephoighygt= a cater naa PFA 1s
activated (beeneate hence gyros is active, mere attr 61
• Fasting state : Low insulin glucagon ratio. Phosphorylation of PFK a occurs and
fructose 4, bisphosphate is not formed,
Hence PRI | remains inactive which inhibits glycolysis. a. Allosteric regulation
Biochemistry v4.0 - Marrow 6.02022 on1200
57s dry EE pd dds emapees dc ut loss, of consciousness due to bleeding from a stomach
ulcer. er respon te ra cpr oP se as od edi 1. sid Nl Send for lab investigation.
why is here loss of consciousness?
fos Aerobic odation of glucose afSected which the primary metabo Suel for brain.
Brain camnot survive f glucose i not going to oxcative phase, resulting o oss of
consciousness.
The hypoxia induces he gene transcription of enzymes of which metabolic pathway?
fin: poner cusces arsrprion Factor | GIF D wil crease expression of enzymes of glycols.
Q Lack of hie pathay results in haemoss. wh?
Ans. RBC solelt on anaerobic which if eta oan to dR RBG SPIES
Q.86C need ATP +o maintain on gradients in the membrane. The lack of i results in swelling
of 6C and lyss. To maintain the ion gradient the major source of ATP & which pathisay?
Ans. Anaerobic gjyeolyss.
@ Enzyme defect n gycolyses cause muscle fatigue. why? fos. Exercising muscle in hupoxic
sate depends on anaerobic ‘gueoiysis. Lack of ATP production results in muscle fatigue.
@ Heart © susceptible to hypoxia but not skeletal muscle. why? fos. Heart has very low
ghycokyte capacity and hence &
susceptible 0 hypoxia. This when compared +o skeletal muscle that has high glycoltie
capacity and switches over to
‘anaerobic pathisauy fo ATP production.
Biochemistry - v4.0 Marow 60-2022 94 Carvonyarates
15
APPLIED ASPECTS OF GLYCOLYSIS
igolysis and cancer ©
Worburg hupothesis
ven by Oo Warburg in 93s.
In cancer cells, glucose uptake occurs at a very high rate. aod astaconuasiad to lactate. This
conversion produces far less ATP (a ATP per cule) as composed 4o glycolysis (32 ATF),
Inorder to compensate this ATP
deReiency, cancer cells fake up more gucose.
In cancer cells, there lactate accumulation which creates an acidic environment.
Lastate needs to be regenerated to ghucose through gluconeogenesis, needing more ATR: i
Therefore, cancer cells go into hupermetabolc state.
This leadRo ince @skheREN 10105
metabolic reprogramming :
Normal cells.
P04 > pep > Pyruvate > Reety) CoA-> COR + HAO ete,
Pyruvate Kinase & in tetramer state (high catalytic state) which s pyruvate Kinase mi. when
#his enzyme s active, | pyruvate wil go in oxdative
|pothwoyss.
Cancer cells derive energy from aerobic ghycolyse.
Blochemisiy v4.0 Marow 80-2022 a Applied 10 Jolie Corer cts ce egos sed Ts phen Fc
Concer cells induces WF |
Lo
(increase transcription] [Increases transcription
of enymes of | [of vascular endothelial Juco Growth factor (vese)
fngiogenesis.
Glycolysis & radiology 001000
a Puorodeowy glucose (Fde) (7 labelled glucose) s gen to locate the cancer.
@marrowedition6notes
Fda injected into the cell > Fda, preferentially taken side. Hhrough GLUT transporters >
Glucose consentrated inside cancer cell 10 times > Labelled glcose undergoes frst step oF
glycolysis» Hexokinase step > PhosphoSuorodeoxy gu- cose Recumulotion in cell-> Decay >
Emits positrons ‘Detected uth PET scan.
Chemotherapeutic agents under tra :
Inhibitors of glycolysis used to Kil cancer cell.
• 2 deoxy gueose (308) hibits golf pathisay ot the level of hexokinase.
• Londomee.

• 3 bromopyruate. i

Biochemistry - v4.0 Marow 60-2022 96 Carbonyarates

15 metabolc defects in ghyeokyhe pathunyy ©
Pyruvate kinase deficiency: +a mV enzyme deflect in humans.
• Defective pyruvate Kinase > Anaerobic goss affected > No ATPS > lon channels not
maintained > ca™ influx + 1" eFSiux > Swelling of RBCs > Hemolysis — Hemolytic
anemia.

• n26Cs, 32 66 creased > unloading of oxyaen (compensatory mechani).

a. Aidolase A defect :
• Hemoiyss.
2 muscle PPK (PhosphofructoKinose D deflect + + exercse intolerance.
0623
iconind RBH QL GEE ERGs i has bow o8evty ans igh tm value for substrate glucose.
Whenever there i high gucose, # is sensed by gicokinase, here the glucose enters the
pancreas trough GLUT-3,
‘gets converted to a phosphate + AT. This building up of ATP ‘enables the release of insulin
from beta cels.
mony a: Giucokinase > netic property altered > Guiding up of ATP fot seen > Camot secrete
insulin even in huperglucemic: state.
Biochemistry v4.0 Marow 60. 2022
PYRUVATE DEHYDROGENASE
Fates of pyruvate 000156
_— Pyruvate Dehydrogenase ei 0 edition Bnotes + site mitochondria. + Formed as a result
of golusis (n cytoplasm). + Proton sumporter transports pyruvate from cytoplasm to
mitochondria.
POH: Multienzyme complex
3 enzymes Scenes “£1 pyusate dehydrogenase | + Thamine Pyoghorphate (TPS) + £3
diukoipoy wansacebase [+ Con
pede o drgtoipande ro dengerase nr
• This multienzyme complex i similar to other multienzyme complexes + I. a-
retogutarate dehydrogenase complex | & HGH
(rea cup. a Brondhed-chain keto acid dehydrogenase complex
Blochamisy v4.0 Marow 60-2022 (@CroN) used for 60AA Catabolism)
All require all the § Coenzymes.
, and, are also required for (-OGH and BCHOH. metabolic defect in &, or &, => POH, @-
HKDH and BCAOH will be affected.
They catalyse Oxidative Carboxyjation
Pyruvate dehydrogenase reaction wo9ss
ote GO oa]
‘ach NADH produces 45 ATPS and each cycle produces 3
NADH 05 glucose s a. GC compound. total ATP produced 6 5 arp.
Blochamaty - v4.0 Mamow 80. 2022 1
Regulation of pyruvate dehydrogenase 01526
This enzyme active in well fed state > under influence of igh insulin-glucagon ratio >
Dephosphoryjated POM (tive. state).
esse: pra.) res 6 8 a freien
Increased acetyj Co : CoA ratio
Signifcance of pyruvate dehydrogenase : reverse reaction L Pyate ———————— acetyl
con a. No enzyme in human body con circumvent ths reversible enzyme. fcety Co camo be
converted into pyrusate. Aety Con s never a substrate for gluconeogenesis
Fatand glucose @marouediionfnotes 02128
3. Fat (Tri Rey GlyceroD cannot be converted to glucose.
• a exceptions: Udycerol
i. Propony Co (Formed from odd chain Fatty acid).
4. excess carbohydrate is stored as fat in adpose tissue.
ae a Gua Ale = Reg
5. In chronic alcoholics > Energy depletion (ess ATP producton).
• Reduced absorption of thiamine (co-Sactor) = Less ATP.

• Nutritional deficiency (8 complex vitamins low 8), 83, 83,89) > Less ATR

Blochamsty v4.0 Marow 6.0. 2022 100 Carbohycrates.

16
a. cetyl Con enters TCA cule.
TAC em ee Pow) Low reaction. Clinical significance w2920
metabole defect (POM Complex: & MC affected) + No production of feet) Con =» Increased
puruvate Converts to lactate > Lactic asidoss. * All oxidative pothisaus cut down > Brain
afSected (dependent on oxidative pathway) > Neurological maniSestations (psychomotor
disabifty).
@ A male chid presented with profound psychomotor disability, a metabolic disorder due
to less utiisation of
puruiate, which resulted in accumulation of lactic acid. ATP production is stopped. : *
which@neyme wmples saffected 7
fos POH complex.
Q. excess carbohydrate leads to excess fat positon.
fins: Al excess carbohydrate are pushed through valve lise Pom, irreversible §9eebIE
Q. Fatty acd cannot be converted to glucose. fins: Because of irreversible nature of POM,
and there is no enzyme to circumvent the POH
Qin chronic alcoholics, here less ATP production fins. There is energy depletion due to
nutrition deficiency, and the & complex wtamins are required for POH and
a-wetogutorate dehydrogenase.
Blochamisy v4.0 Marow 80 2022 GLYCOGEN METABOLISM
Structure of glycogen 000113
eranched polymer of alpha D glucose. Linear side : 1,4 glycosidic linkage. ranches 1 @ 1,
Inkage.
Siyeogen stored in eyfoplasm as cytoplasmic aranules, in the form of @ rosettes.
£och alpha rosette > 30-40 f particles > each is &1 om in diameter + 55000 residues + 200
non-reducing ends (no free. Sunctional group).
Free functional group present only of "glucose residue linked. +o polypeptide © iycogenin
(ony reducing enc).
Reasons why glucose © stored as giyeogen'
ro @marouedionsnotes
• muitple non-reducing end. be released ot a foster rate).

• Low molarity of gycogen.

1 gucose is stored as # is, high molarty will be seen, which attracts more woke, leading to
cell swelling and death.
Classification of glycogen metabolism ~~ ovo7.18 Gigeogen synthesis:
Process happens in Fed state (4-G hours postprandial state). Shucose > Gicolysis/ glycogen
There is high nsuli-gucagon rata In a basal metabolic rate, it excess gucose if gen, it
stocedloa giogen.
Shycogen supthesis
Sie + Liver 00% by weight) muscle (1-3 by weigh. vighest content of glycogen present in
Muscle.
Wafhest # by tse weight: Liver
Biochemistry - v4.0. Marow 60-2022 102 Caotyarates 17 organelle of ghycogen synthesis:
Cyfoplasm.
Steps: hyeogen sypthase (rate Imitng enzyme). eranching, © Synthesis of uoP glucose.
+” ube gucose : Retive glucose donor.
• Nucleotide involved : UTP (uridine trghosphate).
lucose (6) are ) exckinase op
Glucose 6 PO,
[reps
clucose PO, * aseegdoalippéprosshonyose of
oP ghcose
2. diucogen synthase (ate Imiting enzyme). «Primer + igogenin (tyrosine residues -
poupeptde).
• 1-8 gycogeni residues added Guihout enzyme).
• Syptnesis of near chain glycogen.
• No enzyme required for nial addition of chains. on pobymerase and Glogen sunthase
both need primers.
Cohpraonnd 4 lewage up fo Es poy Gucose rescues near pose addon)
Biochemistry - v4.0 Marow 60-2022
— i — + ranching enzyme (alpha 14 1s glucan transferase).
• Mexasaccharide residue cut from end and transiocated to another location in the
polymer.
Glycogenolysis mes
Occurs in fasting state (post absorptive stote).
1n early fasting state (4-1 hours after Sood) Source of blood gcsse Hepat BEEBEo Lc
vormone* Gucagon.
ow sul-gagon rf
She Liver, eletal usc.
In cytoplasm, lysosomes 1-3, Se plays a rok i Iver.
Steps: .diycogen phosphoryfase (rote limiting enzyme) : Needs PLP (amin 80) __ iyeogen
phosphoryase 0© Les a $
a veoranenng enzyme. + fipha 14 14 glucan transferase 1 alpha 1 gcosidase. 5
a an # ip, oii Cs fe
Lekage
ae Hag
Biochem 40 Manow 60-2022 104 Carbohydrates
17
@y the action of enzyme alpha. 14 glucan transferase, the ‘terminal 3 glucose molecules are
eut and relocated to non reducing linear end
8y the action of enzyme alpha Yo glucosidase, it cuts the terminalglucose of the branch
from Ile Inkage ond releases as free ghucose.
23. Conversion of gucose-I-PO4 to ghicose. inter: 1 P0, | Prosphogbicomutase CPemn]
6.60, present in eytocol and traneported to See va 71 Ho | seat tted pnt nse ° elucoce
Gormed in SE2 § released into cytosol va. 71 and Ta)
in musa Marrowedition6notes
ro 2, eypo, —-—> Pyruiate > Lactate
Pracrobic glycolysis (due to absence of
Gop in muscle) ATP is released. Net 3 ATP
hexokinase step is bypassed).
‘The role oF glycogen in muscle is to provide ATP for the: muscle itself. n stressful situation
when epinephrine acts, oF during exercise, glycogen converts into GiPO,then PO, hich
undergoes glycolysis iucose 6 phosphatase + + elucose-- phosphatase activity seen within
the smooth ‘endoplasmic reticulum, + enters Se2 through TI transporter and exits into
eytoplasm through Ta and T3 transporters. + Glucose & phosphatase not present in skeletal
muscle and adipose tissue,
Biochemistry = v4.0 Marow 60-2022
Regulation of glycogen metabolism : 1. Hormonal regulation. 4. Allosteric regulation
Hormonal regulation : Fasting state/stress/exercice +
• Retive hormones + Giucagon (iver). epinephrine (iver, muscle).
• more glycogenoiyss, less lycogenesis. ‘dlucagon § epinephrine activates 6 protein
coupled receptors hich actveates @ orotein > increased adenyiy) eyeiase.
1 SK came J came dependent protein kinase A ‘aiyeogen phosphoryjase elysogen synthase
(Phosphorylated § @marrovipraspbat patocks vce ptt becomes inactive) eiycogenolyss
active, edyeogenesis inactive.
Fed state: + petive hormones + Insulin. + more glyeogenesis, ess lyeogenclyss. * In fed state,
insulin activates phosphodiesterase ‘enzyme, which converts cAme to s! AMP. Hormone
Insulin ATP —><—>came 4 serme dependent protein kinase A Insulin activates enzyme
phosphatase
flyeagen prosphoryjase —_eiyrogen sypthase
Gephosphoryated § Gephosproryjated 4 becomes inactive) becomes active) pecan
ceapecawauwe
‘Biochemistry v4.0 « Marow 6.02022 106 Carbonyarates 17
In muscle vara
1. Epinephrine -> cAMP dependent protein kinase A acte.
a. Nerve impuise (sress/exercise) > Caicur released from sarcoplasmic reticulum >
Increased ackiity of calkium calmodulin dependent protein Kinase > Sigcogen
phosphoryose active > Giycogendlys
3. Extreme anoda (strenuous exercise) > myosin ATPase release SAMP from ATP > Binds
to site in muscle. hgeogen prosphoryase > Enzyme becomes active without any active
choschoruation Samp i allosteric actor of glyeogen phosphoryjase.
Allosteric regulation : In ver ©
Spgerois | eiypogen | Sogensupthess @maloyfditionsnotes | ahs © | J¢&- euro,
os / 00.” quiose 1 prospiate
| © spo, ° x / @- Sustrate
Glucose 1 phosphat @=jproant 3 Glucose in not a. product of gucogenokysis in the muscle,
hence it is not an allosteric ackator in muscle.
Blochamatry v4.0 Marow 60 2022
Questions ¢
@.18 steady state of blood glicose is not maintained, the person wil become hupogjyeaemic
and experience coma and even seizures.
wpe are «8 4oa Salausoaupintain blood glcose and why ? Glyeogen in heart.
& iyeogen in brain.
C-Gieogen in muscle.
0. 6ieagen in ner.
iyengen in he glucose maintains bod gucose for the frst 471 hours of fasting state. By lo~8
hours, glycogen stores are depleted.
Q.1F a glucose load & given 4o a norma person in a basal metabolic stote large glucose is
ingested. what wil this cesult i?
fos. excess glucose frst undergoes glycolysis, then goes for ‘gycogen sunihess and then fatty
acd systhese.
(@marrowedition6notes
Bicchamatry 40 Marow 60-2022
17
wem——— -_rr GLYCOGEN STORAGE DISORDERS ‘Group of disorders associated with
defective metabolism of ‘dyeogen. aljcogen storage disorders (650) can be dwided. into two
+ " iis mea ina Fasting hupoghycemia is seen. Liver is responsible for gjucose levels during
early fasting (4-1ehrsD. No exercise intolerance. a Rest see hw hai ace source of blood
glucose, it onky supplies giycogen for ce or mt wnt i marrowedition =o Ty Type a (von
Gierke's | win hupertrophie | without hypertrophic [ERA heme. [she common 660) tes
Rime fie El ICT aces
ae | aaa (cords disease/ disease) Fob's disease/Limt | prod i pr Andersen disease)
PF Gee -
i (her's dsease)
Biochemistry - v4.0 Mamow 6.0 - 2022 Von Gierke's disease 00:05:03 Type la disease also
Known as von Gierke’s disease.
Biochemical defect + elucose-t-Phosphatase enzyme defect, This enzyme needed for
glycogenslysis in the liver and gluconeogenesis.
@iochemical hallmarks : * Fasting hypoglycemia. + Lactic acidosis + Hetosis, +
Hyperuricemia. + Hiypertipidemia. 4h Normal individuals: Stycogen ‘€lycogen [ -
G@myghespheciase hioles
| Limit dextrin } (@lysogen ¢ short branches } bd ~ vebranching — _/ @lucose - |
phosphate a
In patients of von eierke's disease, the glucose~o- phosphatase enzyme is deficient.
Therefore to produce ‘glucose, the body shifts 4o gluconeogenesis, Gron-carbohydrate
substuserw yor guess paragain fails 40 produce and release glucose because of the glucose-
v-phosphatase enzyme deficiency,
Therefore, gluconeogenesis aso fails and the patient ends up Unith Fasting hypoglycemia.
‘Biochemistry «v4.0 - Mamow 6.0 2002 10 Cameron 1 obae
Pure 11
ao |
Increased uric acid
xcess G-e-PO4 now enters MTP pathway to form purines ond couse hyperuricemia. 6P0,can
oso form puruvate and then feet Col feet] Con can then be used in TCA cule (but fais to do
50 because of depleted oxaloacetate) or For Ketone body Synthesis and results in ketosis.
Reety) Con can also be used for the synthess of fatty acids, resulting in excess riocujayeerdl
and eventually huperipdemi.
Ey
@marrowegitionagisete
eidosis bod Spiess
dy ms epic 1 oC vereasedtrghperde Hperipden Clinical features of von Gierke's disease b
Chitty doll lke facies. a Tho extremes. | i 3 Protnung abdomen i a. Hepatomegoly. b.
Renomegaly (due to increased fat deposition). © bo spleen 4. wypogiypemia.
Blochamisiry - 4.0 - Marrow 6.0. 2022
investigation Increased lactate. 3. Increased uric acd 3 Increased etone bodies. 4. Decreased
bood gucose. 5. Increased triacygycerol. IV Gucagon challenge:
Type lb &so + Involves all features of ype 1a &SD + Neutropenia | aso Seen, resulting in
recurrent bacterial infections.
TypemGSD ooe22
Ro called as limit dextvinoss or Forbe's disease or Cori
disease. were the debranching enzyme is deficient and
hence, mit dextrin accumulates in he Iver, This results in
deposition of abnormal glycogen iver: (@marrowedition6notes
Gogo go Tum dere a1), owas
progr cap pobyrer of gueose | Sipocen : prospronose Clinical features of Type I + Fastng
upagiema. a. protruding abdomen: a. epatomegaly (+++). b. splenomeaaly (9)
e. wo renomegaly. 5 micronodular ner cirrhosis: Geversisle after puberty
investigation: 1 1 no increase in Lactate and uric acid i a etoss.
Liver encymes are elevated.
4. V glucagon challenge can be done.
Biochemisiry + v4.0 Marrow 6.0 + 2022 112 Carbohydrates
There is deficiency of branching enzyme in type IV GSD and albnormal giyeogen
(amylopectin ixe glycogen) accumulates.
Clinical Features + This is a Satal condition +
$ Mupeaiysemia.
• retosis,

• progressive micronodular ver crrhosis which ean cause:

1 Portal hypertension
. Esophageal varices.
3 Uses FAtffé weditionGnotes
4, Death within S years.
Diagnoss * ey bypoglycemia and Ketosis. Note thot in type IV €50, there is no lactate
acidosis or uric acid increase. * inereased elevation of liver enzymes (transaminase). *
Alonormal glycogen which is emyopectin can be viewed on electron microscope.
Type wi eso
Also Knoun as Her's disease. The defect is in hepatic glycogen phosphorylase which
converts glycogen to limit dextrin, Gut gluconeogenesis is intact. Therefore, hypoglycemia
is seen,
bout not severe. ——*— in elycogen- Limit dexter
vee prospharyase
Blochemisty + v4.0» Marow 6.0 2022 1° ce esr orcas
Muscle Glycogen Storage Disorder 002648 —Gintgerpe | eitiattapereaie | cardiomyopathy
cardiomyopatiy
Type eso Type v eso
Donon disease (defect
Is eceorl associ | memiorane protein-a)
Type es: ‘Aso called as Pope's disease.
Lysosomal storage disorder. lysogen metabolem is affected in the lysosome.
Gf Yo gcosidase (acid moitase) is affected
Clinical Features +
• Feeding difficulties.

• Failure to thre. ~@marrowedition6notes

• Hypotonia. Sloppy infants)

• Hypertrophic cardiomyopathy causing carchomegaly and

death (around a years due to cardiac Falure).

investigations
• x-Ray to view cardiomegaly

• elevated serum creatinine Kinase.

• Gevated serum ner.

• Pevated Serum LOH.

Type v 660+ ‘Aso Kroun as meArdle's disease. Detect in the eneyne.

muscle usagen ghosphoryase,
Clinical features. j + exercise intolerance. * Second wind phenomenon : Pain during exercise
subsides with rest and then continue with exercise ‘alter sometime.
Biochemistry * 4.0 = Mamow 6.0 + 2022 114 Carvonyeratos
18
• Rhabdomyokysis myogobinuria.
a. Burgundy colored urine.
Tipe wi eso: Also called as Tarui disease. The defect is seen in muscle § a
scl + xercise ntlerance, mupgioburia, no second wed prencmeron.
engroce: vemeise
most common muse gjyeogen storage disorder in adolescents Type v eso.
Tupe 0 aso: eSect in gycogen syphase. These patients die at very young age as gogen is not
suthesized.
Recently sided Qlseamnstrerandgorders
Fanconi Bickel syndrome : 6LUT-3 defect.
Causes proximal renal tubular acidosis and impaired glucose utieaton
Liver 55 wih myopathy: Tupe Nl and Tupe IV.
Liver 0 with newrdogeal manifestation : Andersens dsease (the bran and anterior horn
cells ave aftected),
Identifying GSD 03720

Sasting hypoglycemia. present then: Liver 50.

8 exercise intolerance is present : Muscle 650.
Fasting hypoglycemia Liver 65D > Abnormal glycogen present = Type IN § IV 60.
Fasting hypoglycemia. > Liver 650 > Abnormal glycogen
absent > Type 11V1€50. ie
nse ilar + red 0 Sasi phenomenon present > Type V &SO. este hare = mud 0 Send id
Phenomenon absent > Type VI G60. eae don tnt kT = i 1 pten ress th gem. toss re diagnosis
can be between type 1 § Il. WW glucagon challenge is used to differentiate. hang hen dry tes
• well fed state.

• Overnight fast. In von eierke's disease, since &-l phosphatase (which & required for
release of gjucose) is absent, there is no rene ho hac mt set

In Coys disease, some amount of glucose synthesis occurs. Via production of gucose-I-
phoshphote by gluse yea TN TERT
increase in glucose in fe. This & not seen in
overnight Fasting as the efSective glycogenosis does not
produce enough glucose 0 cause a. surge in levels.
wo [Von serves cage |
Tupect aso ostect en Lpe0w5 _[sogennpuse | Tipe reo chcose prosgratace Tipe neo rea
matose Type wesc cebruncigeryne i Tipe v cco rancho evzyre i Tpeveso msc prosgrriase i
[po vinan vepate prosghoryase [rope neso | osprotectonmase | Fanaa Sastosl eupvors. Sh
——y |
Blochamisty- 4.0 Marow 60-2022 nec -_r GLUCONEOGENESIS
Concept of gluconeogenesis oor
In diferent stages of Fasting, the body apts gucose by
aemrers Geariy Sasting) Giyeogenohyss (hepat. 10 to 18 hrs + Al gycogen wil be depleted.
o-48 brs (Fasting): Ghaconeoqeness. Supplier of ATP for gluconeogenesis Fatty acid
oxidation (by B oxdaton).

‘Definition of gluconeogenesis + The process by which glucose is sypthesized from non

carbohydrate substrates. + Non-carbohydrate substrates : I. Giucogeic amino acids : arin s
the principle ucaneogeric amino acid. a@sssteweditionnotes 3. 6iycerol park of fat. 4.
Propiony cof From odd. chain Totty acid oxcation. + Note Reet cons never subsite fo
gucanengenesis.
He + Liver (majorky 3 ridney, organelle : Cutoplasm § mitochondria, Smooth endoplasmic:
reticulum (528) also has a role.
I. Coris cyple / Glucose Lactate cycle + Organs involved + Skeletal muscle, Liver, 28C. + Wt
prevents lotic acidoss inside the muscle. * In 2eCs, lactate is the end product of glycolysis. +
1 happens hen muscles are involved in exercise.

— ver /! he muscle [Same (or - ai sais

a. Cab Cuyle/ Glucose Ronin cycle :
• Ws invotved in he ‘stages of fasting.
Liver
ccose ret [ Guzose Smarowe iitioh6notes Pyruiate
Blochamisy v4.0 Marow 60-2022 118 Carvonyantes 19)
— peso ss - i eS , a a
. Succi) Con €———*———— L-methujmalonyj Con = 4 te
Oajoocetate (OAD ——huconeogenesis * In case of vitamin 81a deficienci : There ic
accumulation of serum methymalonie acid.
Key enzymes of gluconeogenesis 01615
L Puyuvate vinase
tae sere] —
• ceguintary st? (mochondriad @ tote aspartate shut To argon 08 across crcl membre.
ps
essen
‘Biochemistry v4.0 - Mamow 6.0 2022
10 =" iets
© pee cortons nse se epee Frosphoena Rywiote dearbonjaton loved & a) oo" Ge)
typgaryaton ae a Apres rts Paes gree ee sh Fe ornate fructose ie erogres reesed Few op —
emrospratinse op fer reversal —_ ener. * Inhibitor of Fr. Yo 6 vrainase anecinase eucase
MPNPESS 60 . core o- freretace ie 6 P0, PES, eucose ‘srequred fr a a wo § ‘Summary of
gluconeogeni tos 002739
(-SUCCIM. Con << Propiony Cot
Pe Peper, ax Lactate —> pywsate “Sonn pep >a pa
vere exe 5 ars we ara sera newse wood acactte —y1 acne ca sere oa 90 se wo Debio, ws
owe non non 1 7 orl wm 4 elucose A eb = PF 6 = PO FO OP i cose
Regulation of gluconeogenesi
Hormonal regulation +
‘locheisry* v4.0» Marow 6.0» 2022 120 Cabonygrtes {0
• Bete in low insulin glucagon rato: Enzymes are in prosphonyted state (active).
Alosteric regulation
• fetal Con allosteric actwator of I” reggltory step of gluconeogenesis (Pyruvate
carbosfase).
• Fructose 3 ieghocghate allosteric histor of Fructose te bighsphatase.
Applied aspect of gluconeogenesis ws0y
Ca sgn
Raw ea © biting
© sete uconeogeness by Ader as # hots
Sufi tonaied fo & oti required steps.
reguate 2 cm ie
© Propany con Carbonose
er hea eonotes
Question I+ Biguanides are used as oral hupoglycemic agent A tneoretical concern about
#his drug s lactic acidoss. what is
he reason for this concern?
focuser : + His because Biguarides nib pywiate carboajase § decrease guconeogeness. +
pyrite s not converted to oualacetate, # may convert nto lesan. + When there is mo 08%,
TCA cue & oso hibited.
‘Question a +A chronic alco brought to casualty, as he was driving heavily for the past |
week and he had rot eaten any Food For post 3 dau. He was confused and was seating
profusely. He developed seizure in the casualty, He blood glucose drawn just before seizure
was 30 ma/dl and blood. ‘ethanol wos very high. hat are the metabolic changes i hin
whieh resulted hopoghyeema? fiver
Blochemity 4.0 - Manow 6.0 2022 hol cetaidende a eetate or don Ea
missense
ogo creased 352% ato ~ (om —>—" molate 1 1 v male, | are von Pate 50m | Gepiton ol ow X
} ct emsiocs
arent © | Quoonogeness
Question 3 + A patient with acute exacerbation of chronic bronchial asthma treated with
high dose IV methyl prednisolone. On discharge she was discharged by switching over 40
oral prednisolone. Abter 5 days she presented win polyuria, ogg, ond muscle sprees, SRE
bos
clinical history biochemically?
Answer : ueocarticods ficken muscles Gama mais
=
jaceases
toto rayne
dee ueonesgeness

5 a result Increased Ghuconeogenesis keading to

dexelopment of given symptoms.
Biochemistry - 4.0 + Marow 6.0 2022 122 Cameron 20) [ome MINOR METABOLIC
PATHWAYS
Galactose metabolism 0220
Function: Convert 10 Glucose and Used in synthesis of lactose, Glycosaminoglycans. Site :
Liver (Major), erythrocytes, Fibroblasts.
S| man mn) 8G; | [Tera
@martchesdiionénotes Galactosemia, 0076
Classic aaloctosemia. Non-Classic ealastosemia. AT defeiency. aloctorinase deficiency: Ory
Clrical cataract,
Biochemical Defect : [ee] [me og fon tt ce TT word
=——g
ot thon rsgrcgace. dans of gucsosogeese. | TT ra Clinical features : Age of onset : “First a
weeks of life” — reast mik (Galactose. present.
lochamisny + wi0 Mavow 8.0 2022 _-— Feeding difficulties, vomiting, Jaundice, Failure to
regain birth
weight.
Hepatomegaly, Liver Sailre, mental retardation.
“Ol drop Cataract’,
Neonatal Sepsis due to eco bacteria,
Lab dagposs + Urine: Reducing substance positive (benedict ted (on speci).
iucose oxidase test: Negatve.
mucic Acid test : white precipitate (specif).
enzyme studies.
‘Genetic mutation studies.
Treatment : Restrict Lactose + Til + years of age pri proce | /1 Sor Breast Seeding, Ce [toe]
Goloctoknase and epimengsie, JERIGERIASR EROS
conditions manifests as Cataract only. Fructose metabolism 629
Source : Dietary sucrose, ‘Dietary free fructose : Honey and fruits, Polyol Podhusoy Gucose
> Fructose. Ste: Liver.
ei ns SEA 124 Carbonyates
20
Hereditary Fructose Intolerance © Deciency of Adolose 6 => Fructose | Phosphate
fecurulotion > Toe.
Simiar to Galactosemia.
Traps inorganic Phosphates -> Inhibits glycogen Phosphoryjase and enzymes of
aluconeogenesis — Fasting Rpogieeic.
¢/F: Simiar to gplactosemia except,
ge of onset : round months (age of supplementary feeding).
Feeding difficulties, vomiting, Jaundice, Faure to regain birth weight.
Hepatomeqaly, Liver Sailure, Mental retardation.
No Cataract.
Lab dagposis
• Benedicts test : Posie.

• Test Rorifieioeasc EMBO les Rapid Furfural fest: Positive. ‘Seluwonaf's test: Positive.

• Guucose oxidase fest: Negative.

• enzyme studies.

• mutation studes.

Treatment: Restrict Sucrose.

Essential Fructosuria 02629
Deficiency of Fructokinase. Benign Condition. Fructose hos no renal threshold > Fructose
excreted in urine. Fructose is harmful because + Fructoknase is rot dependent on insulin
and converts to Sructose-t-phosphate. + Tightly regulated step of goss PPR 1 not seen in
Slochamietry- wi.0 + Maow 60-2022 "20 orm million Fusose table. * cach convert Pit feof
Co attycd > Toe wot
DE" erensed Frustose and igh ustse Supe | [epdaize | digress The oc Hexose
Monophosphate Pathway waose
Other Names : Pentose Phosphate Pathuiay, Dickens Horecker
pathuau, Prosphoguconate pathiay, Shes Cytoplsm, @marroweditionénotes Phases :
Oxidative is Irreversible § Nonoxidative is Reversible.
90 not produce ATR. 0, s produced.
Oxidative phase Irreversible.
Glucose-s-phospate (6) Ls > Glucose-G-phosphate. NORM | (rate taming step)
Biochemical Signcance I. Generation of NROPH.
Giochamiaky - wi0 Marow 60+ 2022 -— 126 Carboryoraes 0)
Functions of NADPH I. Free radial scavenging : Important in RBC and Lens.
re oucdatve ope NOP Phase.
2. Heep ron in reduced state in Hb (Fe) (event
methemogobinemia). 3 Reductive Bosuthess of fatty acids and steroid hormenes.
Organs of HIP Shunt + Liver, Adipose tissue, Adrenal cortex, onads.
Non-Oxidative phase 03908
Produce RRRsAH we dition Bnotes Al steps ove reversive.
eiochemical Significance © rp ope rome ne se mars ioe, en, — =" Em em wm Wn een
3 Vasseur, acon,
‘Biochemistry - 4.0 Marow 8.0 2022 “en cinical sqpifeance I. &6PD deficiency + m/e enzyme
deficiency n Mamas. + X Inked recessive dsorder. + only males are affected.
Aagravating Factors Fava. bean ingestion: Favism. * rugs + Sulfa drug, Primaguine. +
Prevalent in mediterranean and middle east regions (Ove to prevalence of Plasmodium
falciparum).
oF: Decreased PD > Decreased NADPH > Free radical scavenging = RBC membrane integrity
ost > Hemoss = © vemogtc anemia 0) Jaundice. AROPH recived 40 Keep ron in Fe state. In
NADPH defecient Stade H gets converted fo Fe" > Methemogltinema @marroweditionnotes
Uronic Acid pathway oss Oxidative pothusay of gucose. Site Liver. organelle: Cyfoplasm.
Functions: Produces ron ack + Conjugation of Birubin (Gueuronic ace. + Supthesis of efi and
Proteoghycans. © Produce pentoses. © Rscorbic acid: can' be synthesized in humans and
higher primates (ducondlactone oxidase).
essential Pentosuria.
sengn.
Luise is excreted inure.
enzyme: Xtal dehugdrogenase / xylose reductase. neds test: Poste.
Test for pertoses Positive.
Slochemieny + wi.0 + Mamow 602022 128 Carbonyarates
20
05653
To convert Glucose > Fructose.
Ghose, men cy Ty we hr wr Soo
[corey ves an ins: | ur vary and Sem vescls L H Ei [oss chore > Fructose.
denudroqenase less.
| tucose > sorbitol (case ot | Loccumuioton of ducose). l
Q uy Cataract in Galactosemia and om?
aur (Lens) GLUT 1+ Independent of insulin , present in Lens. excess Ghucose To lens
(Adolose reductase present here) > Aidose reductase Activated > Glucose converted to
Sorbitol and eplactose converted to Galactitol or Duleiol (@marroweditionénotes oth
Sorbitol and Galactiol are osmotically actve and attracts 10 > Cotaract
Clinical questions :
@ A person had a road tral accidents legs were badly ojured.. Later i got infected Pus
culture revealed methicillin resistant S aureus . & comination of Trimethoprim and
‘Sulfamethoxazole was stacted.on the. ticd day of therapy she developed jaundice and the
Hb level fal down 10 4g/
dL Suspected an apparent acute hemolysis on exposure to infecton and sulfa. drugs?
hich enzyme defect might have aggravated the attack of hemolytic jaundice and anaemia i
this patient. 3. substantiate the reason for hemolysis.
fos) @ePD deferency, 2 PD and NADPH deficiency
Biochemistry + 4.0 Marrow 8.0 2022 _— a @ A patient wih recently diagnosed Diabetes
mellius darted +o avoid cane sugar from her diet But she continued to consume lots of
fruits which resulted in poor diabetic control 1. which sugar is responsible for the poor
diabetic control? 3. what is the reason 7
fos. igh Fructose may lead to hupertrighyceridemia due fo increased ghcolysi.
@ Aneonate soon ater birth started voy, uncice: and distended abdomen. The consultant
paediatrician noticed that her Iver 5 enlarged. Sooner an ophthalmoloay 1. uh is he
probalbie diagnose?
a. hich enzyme deficiency is suspected?
2. What are the investigations done?
fos: D calactosemia. 2 olactose-i-prosphate dg pnsfemessios ) music acid test (specie) and
urine reducing sugars.
Biochemistry v4.0 Manow 60+ 2022 130 Uds 21
CHEMISTRY OF LIPIDS
Definition : Heterogeneous armel sccmaaunds skis are insole in water but ore soluble in
nen-pdar sovents. Lips ave related to each other more physically than chemically, Bloor’
classification of pics : simple pds © Ester of alcohol (gycerol + acid (Fatty acied. £4, Fos, ol,
waxes. Compound pid + Ester of alcohol + acid + other component. £3; Prosphoipic,
lipoproteins, glycolipids. ‘Derived ids : Derived from simple or compound pds. £9 alycero)
fatty acid. miscellaneous Steroid hormones, fat soluble vitamins. Neutral Fat +
Triocyjglyerol (TAG) is a simple iid. (@marroweditionénotes
° Fatiy acd residue
o00s2
Classification of fatiy acs Based on number of carbon atoms: 1. Short chain Satly acd (SCF)
C,
Biochemisry - v4.0 + Marrow 80+ 2022
& medium chain fatty acid (CAV: ¢, ~ C, 3 Long chain fatty acid (LCF) + 7 ¢,. 4. Very long
chain fatty acid (ACF
Based on the presence of double bond t I. Saturated fatty acid No double bonds.
a Unsaturated fatty acid | (BAdbdeensRsaste mUFR (Monounsaturated FR © | double bond.
PUR (Polyunsaturated FR): > | double bond,
Cl
Saturated Satty acids : [ cr Source | Acetic acid /negar [ Propionic acd GO Butyric acid (40)
Valerie acid (60) id Capric acid (00) CFR Cour 00d (20 nf soa 0 Coconut oi (richest) we Palit
acd 060) Stearic acid (80) foal fet Unsaturated Fotty acids : mar Source a et mustard
ol/grapeseed —— oi (richest source) PUFA Uncle acd (Ca | SafSiwer of Gehesd double
bord) 6A: Indlenc acid (82) | Oi of evening primrose Cindenc acd (6 | Fasseed oi (iehesd |
feachdone acd Go) | oral fat Terrodonc acd Go) i ©: eeosapentaendc i acd Fish ois, algal ois,
¥ Cervonsc acid (33©) breast mk. ore ocosahexaenoic I Lo I
Biochemisry + 4.0 + Marow 6.0 2022
gem A Highest concentration of PUFA is present in sa¥Siower ol. a highest is in sunflower
of. + Least concentration Coconut ol ‘Essential fatty acids : Camot be sunthesised in the
body, + Under acid + aLindenc acd ‘Sem essentil fatty acid : Can be synthesised from
essential aces. * frachidonic acid ( from linoleic acid). * ¥ Linolenic acid (from lidleic acid).
Omega (uw) classification of ttyacids — wine Carbon atoms are numbered from the
terminal methyl group. The fatty acid is classifed based on the position of the I double bond
from terminal methyl group. Sample , fatty acd: Wi oa 3 as 6 1 oe Ci, —CH, 0H, Cn = C= Cn, —
Cx, — COD @mgtrowgtiitioffenofes FA 5 i Anumbering system : Numbering starts from
functional group. ©, fafyocd _ Totty acid [Apaimencacd ah Tanoodon acd Coolie acd Cervonic
acid frachidonic acid (20 © ©, Satty acids s more harmél thon ©, fatty acids Arachidonic
acid is a source of eicosanoids. (prostaglandins, leukotrienes). + These inflammatory
mediators increase cardiovascular isk § degenerative dsorders. significance of ©, fatty
acids ¢ 7 ceri eos ra i decreased platelet aggregation. Decreased inflammation. Infant
development (oHA for brain development).
Decreased chance of mental ilness.
lochemisny + 0 Marow 60 2022 Gren Re] Decreased chance of degenerative disorders.
(rheumatoid arthritis, alzheimer's).
‘Docosahexaenoic acid © 6 an ©, Fatty acd Source : ereast mk, algal oi, sh ois. Uses:
• infant and foetal brain development.

• eetinal development. Low DHA is associated wh retinitis pamentosa. Transplacental

transport is possible for DHA 50, taking OHA tablets during pregnancy may be
beneRcil.

Cis and trans fatty acids wozess

Fatty acids wih a double bond exists n a isomeric forms. +s form: The structure has a bend
(angje + 120. increases Suidity of plasma membrane.
• Tre fom The shat RS (ETE:
‘Sources of trans fatty acids + Vonaspati & the richest source. Partially hurogenated fat
(margarine dalda, vanaspati cake butter) used i bakery products 40 improve she ie. Deep
frying eg french fries. Reneating of vegetable ol
Heat converts cis form to harmful trans form.
Cay allowance of trans fobty ass 3-7 o/doyy
Dusdertages of rare fot ac essential fobty acd defiiency
• Lipd Fractions | increased TAG { LoL, decreased HoL.

• increased cordovascular rk.

• increased inflammatory response.

• Couses insulin resistance.

Coconut oi can be better than sunSouwer of as

• Coconut ol is Saturated FA, trans-isomerism occurs in UFR.

• mCFR i directly absorbed from portal vein.

Biochemistry + w.0 + Marow 8.0 + 2022 134 Ups 22 [i ersas SPHINGOLIPIDOSES

Phospholipids 0c
Phospholipids are compound lids that consist of
Fatty acd,
a fleohol
2. Prosphoric acid.
3. Base : Kirogen containing and non-ritrogen containing,

General structure of phosgholpid :

ester ievage vagdrophole tad consists of fatty acd
trogenous/oon vagenous base.
(@marrowedition6notes Class feation of phospholipids.
oe a ard Ene'e he acsbond
Sphgyin Tm Hsbrpcmn conkalning Hon clam sonkaiy; Cen che Ophespratiy ducers
Prosphats serine faa Phosphatidic acid, lecithin, cephalin 000s } Prosphatidic acid: Simplest
guperophospholpic. lerol + 3 fatty acd residues (acy dycerad + PO,
Bochemisiy - v4.0 - Mamow 60 + 2022 2 iio
No nitrogenous base : o
n on-0-c-g
oo on-o-bog, |
on, -0-fo,
A gyeerophophalpids are derived from phosphatidic acid Diacy glycerol + PO,» nitrogenous
base = Lecithin
(phosphotidy choline. Signifcance
most abundant phosphoip in cell membrane. major constituent of ing surfactant. Store
house of choline.
cephalin: Phosphotidic acid + Nitrogencus base = Cephalin (eth@olarnmed o i(frosphatas)
ethandamine). Significance : Blood coagulation.
Cardiolipin, phosphatidyl serine & phosphatidyl inositol - 00806 Cardioigin : Dphosphatidy
dycerol (phosphatidic acid + Glycerol + phosphatidic acid. Significance © olated frst from
cardiac muscle, hence he name. Present in the imer mitochondrial membrane.
only ontegnic prosghaip.
Disorders associated with defect in cardiolipin are © 1 1. Cardoskelotal muppasty (Barth
sypdrome).
oe i 5 potrgidem
4. veart failure.
Biochemistry + v4.0 Marow 60 2022
Phosphatelyl serine : Phosphaticic acid + Nitrogenous. base (serine). Significance :
mediator of programmed cel death/apoptose.
Phosphatidy inositol significance
cet sgpaling
mediator of second messengers in hormonal potas. sphingomyelin 12s Oni phospholipid
with sphingosine as backbone.
Amico oleohol. ‘Derived from serine.
sphngpsee: © i
@marrowedMbii ffrotes Fatty acd - con Sphingosine + Fatty fed + PO, +NitTogenous base.
Ceramide : Sphingosine + Fatty Acid.
sphingomyelin : Sphingosine + a Fatty Acid + PO, + Choline. Signicance :
1. Outer membrane of plasma. membrane.
a. specalied structures in plosma membrane + Lp rab.
3. Myelin sheath of nervous tissue.
Glycolipids 01523 Complex fi that contain carborurate but 00 phosphate group.
Contain sphingpsine.
Wio/a. ghyosphingplipiis.
Blochamisky - 40+ Marow 60 2022
Structure of glyeotpids + Sphingosine + Fatty eid + Carbohydrate. Three types:
Cerebroside, globoside & ganglioside 00:35:28
Cerebroside : Ceramide + monosaccharide.
| the monosaccharide is + elucose : Giucocerebroside. Galactose : Golactocerebroside.
side prese PMALOWSAHgnsnotes
alactocerebroside : Present in neural tissue.
‘Globoside + Ceramide + Disaccharide/slgesacchoride. £g, Loctosy ceramide.
eangioside + Ceramide + Oigosaccharide (ANA). NAAR: N-Aeety! Neuraminic Reid = Sialic
acid, Namedos 6m, ii L|
L| . Gorgon | unaue ruber: esgred based on raratayogy Monosialo containing
‘em Ganglioside thot acts as receptor for cholera toxin in human Intestine.
‘ems: Simplest ganglioside.
Biochemistry « w.0 + Marrow 6.0 + 2022
2 illite 138 Lids 22
Sphingolipidoses 02253
Definition: Springolpdoses & a group of kysosomal storage disorders charcterised by an
inherited deficiency of ysosomal hydrolase, leading to an intralysosomal accumulation of
sphingosine containing ip substrates.
em gangfosidoss * Case s 4 month oid infant presented uth frontal bossing, depressed nasal
bridge, long phitrum, low set ears. Fundoscopy shows cherry red spot,
Biochemical defect : Defect B-galactosidase enayme
vesutts HABIMAMGH HH Bose.
em gongjosde” S52, ean, argos
Cinical features
Blindness.
Typical Sasies : Frontal bossing, long phitrum, depressed nasal ridge, low set ears.
maculor cherry red spot (seen n approximately Som.
fogokeratoma.
vepatosgienameagiy hentol retort:
em, gargjosidosis Case a year ok boy who s hypotonic, decerebrate and bind, Funds
examination reveals cherry red macular spot. hot 6 the defect in this hid?
rs. Defect in hexosaminase enzyme
Sochamieny- vi Maven $0 2E8 22 spr J - Y -
Siochemical defect : Defect in B-hexosaminidase enzyme
results n accumulation of am, ganglosde. B-hexcsaminidase
am, gangjeside = em, gangloside. B- hexosamindase sotorms a iand ip af Tupes of em,
gongesidosis Tou Sachs dsease Sandrots's deease B-hesamindases | p- hexosamndase f and
8 (deflect nt subur®d. | (defect in subund @rharrowedition6notes | Clinical features.
Clinical features Cherry red spot i macula. | i features of Tay Sacks ard retina. dsease along
uth Vuperacusis hepatosplenaenaly cardiac decks, | abnormalities and bony macrocephoby
deformities. Krabbe's disease & Gaucher's disease wos
5 month od gl presented uth developmental delay, At 9 mois the same chis condition rapid
deteriorated. Now she & in ophothotonus posture usth clenched St
Biochemistry 4.0 - Marow 80 2022 140 piss 22
Frabbe's disease
Bochemical defect defect n f-galactocerebrosidase/ ‘plactosidase enzyme resus in
accumulation of B-galoctocerebroside.
alactocerebrosdase B-galactocerebroside [Brapincioe 2% cerebroside
major location of B-galactocerebrosde in neural tissues. Cinical features
Severe neurdiogical def; No hepotosplenamegaly (as enzyme nok present here) Cherry red
spot + enlarged macrophages causing goood cell
inclusions (in white matter),
Gaucher's disease ©
Case +3 year od chi presents wth abdominal distension 5 bone pain xray of femur shows
peculiar deformity BA Shows cells uth crumpled tissue paper appearance. edition 6oTES
most common osama storage disorder. Biochemical defect Defect in B-gucocerebrosdase/
B-ghucosidase enzyme results n accumulation of B-gucocerebrosde.
-gucocerebrosidase, B-ghuoocerelrose —P-he0eTEbIOSASE, (py Location of -
glucocerebroside : extra. neural tssues.
Clrical features Pain in long bones, and pathological Fractures. Haematological features
include decreased hrombocites §
Biochemistry v4.0 - Maow 8.0 - 2022
2 rite
pancytopenia. This can lead to bleeding manestation § anemia. No intellectual deficit.
No cherry red spot (exception: Tupe I Gaucher's disease - poeuo-cherry red spo present.
X-ray features erermerser Sas deformity.
one marrow shows Gaucher cells crumpled tesue paper/ wrinkled paper appearance).
Treatment eT enzyme Replacement Therap) © Recombinant acid fb gucosidase
(mighucerase). Velagucerase 0. Tolgucerase o. 3. Oral substrate reduction therapy Miglstot
ns. glucose ceramide suphase. 2 Bone marrow transplantation | coushers disease ¥rabbes
dsease Broheoerebusdase [Baacieereorosdase eal eirolation
acGumulotion in non- nae tssues
tissues
omental retardation or | Severe neurogeal deck neurobogjeal deck |
opipenreplye|iohepigerenagl resent
Niemann Pick disease, Farber’s disease & Fabry’s disease ~ mewn
Nieman pick disease :
Biochemical defect : Defect in sphingomyelinase enzyme, results in accumulation of
sphingomyelin.
Clinical Features:
Cherry red spot in macula.
2ebra body inclusions.
Farber's disease :
Case +18 month oid chid presented with painful jont swelling nodule.
Blochamistry + 14.0 Maow 6.0 + 2022 142 Lice
22
Biochemical defect of acid E ceramidase enzyme.
Clinical feature : Painful pint
sueling (resembles rheumatoid arto).
Fabrys dsease : X-linked recessive disorder. males are affected.
Biochemical defect of G-galactosidase enzyme, leading to accumulation of
gobotriaosyjceramide.
Clinical features fngokeratoma. Corneal lenticular opacity,
Falory’s criss (agonising pain in the proximal joints. Hupohudrosis.
urinary sediments show maltese appearance, due to lp nclusior 1
ig dition6notes
cross,
Treatment : enzyme Replacement Therapy (ERT): Recombmant a-gplactosidase
© Agplsdase B Gabrazyme.
a. Agoleidase GreplogaD. Wolman's disease & gomorabfeaturss ofall sphingolipidoses
002551
wiiman's disease 50 known as chesterol ester storage disease (CES)
Biochemistry + v&0 - Marrow 80. 2022 2 gitates
Biochemical defect of acid ipase enzyme, leading to accumulation of cholesterol ester ¥
triacyjghyeero, fey ein ak aap tape
Clinical features + Wotery green diarrhoea. Failure 40 thre, ees,
Soca ea eames,”
General Features ofall sphingolpidoses A ore autosomal recessive, except Fabry disease,
fl have mental retardation, except Gaucher's disease.
fii have cherry red spots, except Gaucher's § Fabry}s disease, ‘Sphingoipidoses with corneal
clouding : Fabry}s disease 4 em, ganghosidosis.
‘weusion bodies: = @marrowedition6notes loboid cell: Seen in Hrabbe’s disease.
Zebra. cell : Seen in Niemam Pick disease. oltese cross n urinary sediment Seen in Fabrys
deease.
Lysosomal storage disorders Disease enaye detect
‘67 ganghosidosis B-galactosidase
Tay Sach's disease B-herosaminidase &
Sandho''s disease Behexosaminidase A 6
|rrabbe's disease B-gplactocerebrosidase
[Niemann Pek Type sphingomyelinase
Gaucher's disease [@ucocerebrosidase | i
Metachromatie leukodystrophy | Aryl suifotase A {
Farber's disease _|
woiman's disease
Fabry’s disease
Biochemistry * 0“ Manow 6.02022 144 Unies 23
OXIDATION OF FATTY ACID
concept + arly fasting (4 brs): lycogensiyss. + Fastin (lo-8B hrs): eluconeogenesis Fatty cid
oxidation (provide acety) CoA and ATP to activate
_guconeogenesis). *+ Prolonged fasting (a days): Fatty acid oxidation retone bodies
synthesis) ‘Types of fatty acids oxidation on0ase o/c oxidation: B oxidation.
Tie fatty acid to undergo Fatty acid oxidation : Saturated, Fatty acid (palmitic acid C1.
Other + VLCFA ( -chain Fatty acid). Aaecabadi anya Odd. chain FAL si exoua xay Ot
oxidation. meat B oxidation 0:05:09
A process by which fatty acids are successively cleaved to AC acety) CoA and release eneray
“The process is Known as B oxidation as B carbon (CH,) group gets oxidised to CoH.
(Blochomiety - w.0 « Manow 6.0 +208 on 72 lites site of cleavage : Between o§ B. Liver,
muscle, and adipose tissue.
in mitochondria.
Steps of Fatty acid activation: D Retivation of Fatty acids. @ Transport of actwated fatty
acid from eytoplasm to mitochondria. D oxidation,
fctvation of Fatty acid ste cytoplasm.
fey Con synthetase Fatty acid ey! CoA e008 / °
TP ame ll
Ga phosphates @-C~ Seon
arewsedtrowedition6notes ‘The only place that requires energy,
‘The enzyme (acy Cah synthetase) stuated inthe outer mmochondrl membrane.
The reaction is happening in the cytosol. ‘The enayne belongs to the ligase.
‘Transport of fatty acid to mitochondria: Occurs with help of carnitine (FA < 4c. does not
require corritine).
capecee
fey crntne carntne Reycariee Len
iochernety « vA + Mawow 60-2002
146 Ups 23
CAT 1 Carnitine Reyhransterase) : Transfer carte to the octwated Fotty acid.
©eactions of beta-oddation ° I B fey con R-C-scoh C=
icy Con dehydrogenase.
ERO, B
Transenoy Con c tae np on
ac
LY
Ga ne =
| cc Ba rE on veto
ec a seni he a 1
RoylCon fcety) CoR (a0)
Carnitine (B hydrox gamma. methyl ammonium butyrate) + Lugine 15 adenasy)
methionine is evolved in the syntheses of carnitine.
Vitamin C is also involved. FA < HC doesnt reauire carnitine Sor transport.
Energetic 2348
No of beta, odaton = No of C atom - 1 § a i No of acety Cof = vo of C atom a Example + Paimte
acid (0). No of beta. oxidation = ko = 1 = 1 a
Bochamisny «v4.0 Nasow 8.0 + 3032 a =
No of acetyl CoA = 6/2 = 6.
1 beta, oxidation produce | ADH, (15 ATF) 11 KADH (25 ATP) =4 ATR
Total = xb = a8.
8 acetyl Con: 1 acetyl CoA produces 10 ATP in TCA. acetyl Con = 0 ATR
Total ATP = 80 + 26 = 08.
For actation of acy Co synthetase, a ATP are used. Net ATP = 108-3 = 06.
eased on dd cakulaton : | FAow, = 2 ATR 16RD = 3 6TP, 17cA =a ate.
The ruumber of 6TPS produced eta pedaten st tear
acids 20.
“The number of ATPS produced by beta oxidation of paimiti: acid is a0.
Regulation of beta-oxidation 002939
Rate Limiting Enzyme (R18): CPT 1 (o/i/a gateway of beta oxidation).
In well-fed state + Insulin: Gucagon ratio 6 high Increased cety Co carboase (acne). malo)
Con ibis CPT | > FA oxidation il rok occur
1 fasting state : Insulin © Glucagon ratio lou. Decreased acety Con carbongase (nactie).
Malon Cof level wil be less —— FA oxidation occurs
Blochamitry + 14.0 Marrow 6.0 + 2022 148 Lipids 23
Oxidation of various fatty aci
wer: modified beta-oxidation. Fore, Ste: Peroxisome. mitochondria also have some role.
Products Acety) Con, H,0, oetanoy Con Octanog Co enters mitochondria. and goes Hrrough
the same as beta-oxidation.
@marroweditionghoted og, Normel ocaton - as co Lb dengue NS Nesom F40 acy) Con
dehudrogenase & bupassed ATP generated 15 less for every double bond in an even
positon.
0d chain fatty ocd +
Reety) Con + Propiony CoA s produced.
EE i — sl Site + endoplasmic reticulum § peroxisome. For branched-chain FA with a branch
at b carbon. mV + Phugtancy] Con (dairy products, green leafy vegetables)
© oxidation of fatty acid Site S68 (ricrosome). No ATP is generated.
AN i NA ecarboae acd
Medium-chain acyl CoA dehydrogenase deficiency (MCAD defect) war20
Q.This is a | year oid chide sibling din Be! aus of Fe of S10. 8 mons she hod. a We-trreatenng
episode of seizure and hupoghycemio. Her blood examination revealed C, dicarboxylic
acids. No Ketone. bodies. wer doctor advised her mother to give her frequent meals. with
high carbohydrates and low fat.
© oxdaton Medum Chain Fobty acid ——————— Dearboxjic acd ¥ B ocdation |
Reetyicon | ATP RE)
1 ® Fess Ae x I | Noketoss Fasting hupglycemia (sudden deat)
al Ta ce 150 Lice
23
@ Biter ingestion of this fruit, man develops sudden onset vomiting 3 hours later; Sollowed
by convulsion, coma. death.
A Jamaican vomiting sickness. unripe ackee fruit —> Hypoglyen
1 © feu Con Dehydrogenase (B oxidation) meter jmp | i | Wetoacid synthesis |
Gluconeogenesis Fekiee fruit is Sound in west Africa (iamaica).
Clvcal features :
RIERA TRMRETa soma and death. Death con occur as there is deficiency of Ketone bodies
along with ghucose, causing lack of subsirates for brain.
Refsum’s disease wsoss
Defect : @ oxidation. Prutanoy Con hydroxjase (phytanoy] Con oxidase) is defective.
Clrical features; eetivti pgrentosa. chirps.
Peripheral neuropathy Cardiac arthymma.
Treatment este dairy products ond geenlealy eapoties
Slochamisty + vi.0- Mavow 80-2022 onan Zell weber syndrome/cerebro hepatorenal
disease o0s007
Peroxisomal targeting disorder. The synthesized peroxisomal enzyme in the endoplasmic
reticulum reaches the peroxisome with the help of PTS (peroxisomal targeting sequence).
mutation in PTS will lead to no enzyme in the peroxisome. LCF oxidation and t oxidation
affected.
Clinical Features :
mongplod facies. vypertelrem. Unslanting palpebral fissure. Frontal bossing, igh forehead
erushield spot in ris epicanthal fod. Resembles down syndrome. Daagnosis: Peroxisome
ghost. fecumulotion of VLC in the peroxisome. fecumulotion of phytaric acd.
@marrowedfionenotes
Ciical problems Reason for Sasting hypoglycaemia in fatty acd oxidation disorders ?
ole of fatty acid oxdation & to provide ATs are guconeagenesis. Once ta hours of Fating is
over, body
ets glucose from guconeogeness. Hence when the fatty ‘oxdation & blocked, giconeogenesis
does. not occur leading 6 Fupoch rail
why drugs that reduce fatty acid oxidation are oral hypoglyeemic agents ?
example: Suhongoreas.
In om, there wil be excess gucaneogeness So oral
upoglgcemic agents blocks his excess guconeogenesi by ichioitng fatty acid oxation.
Slogemisty +940 Mus 88-3004 152 Lids
24 Em KETONE BODIES
early Fasting (-1o hrs) — egeogenoiysis. Fasting (48 hrs) — eluconeogeness.
Prolonged Fasting (starvation) (> a days] — Fatty acd oxeloton Hetone bodies. Provide Sue
for vital organs ater depletion of gucose. The rain can derive ony 20% of energy from
ketone bodies.
Ketone body synthesis 0025s
Sie + Occurs only in the Iver, inside mitochondria. fey con Geetycom |B osoton
(@marroweditionBotes feetoocety) Con
coger | He Con Synthase tartng substrate)
me Con (3 dro 3 methujghtary Cod feetycon_| Wma con Lugae
2n
were fronsutyote
Starting substrate : feetoacetyl CoA. Rate limiting enzyme : HIG CoR synthase (tochondria)
Cujtosolic HG CoA synthase for cholesterol synthesis
Aeetoacetote 1° retone body
feetone a° tetone bodies.
B- on eutyrate.
Schamisry va + Mavow 80-3028 2 ee
Ketone body utilisation 0810
Occurs in extrahepatic tissues as ier lacks thiophorase.
enzyme Sor ketone body ubization Succiny Con acetoacetate Cof transferase Thiophorase.
Organs. that never ubiise ketone bodies : Ler. rec. feetone © * Volatie, neutral + excreted
through the lungs It has a fruity smell which §& seen in Diabetic Ketoacioss.
Starvation ketosis |
After wtisation of ghycogen and faiure of gluconeogenesis, Tog becomes the next source.
Tha @fharrowegitionénotes fed aigceral PB oxidation = (oaiacetate Acetyl CoA deeted) oA
Tea cydle Wetone body synthesis
Dabetic ketoacidosis 1 score 4 sin 1 ahose toon AS [> ——seluconeogenesis ounacetate i i
ve
Increased gjucose in the blood but cannot enter cell as insulin dependent GLUT 4
transporter is reduced.
Biochemistry + v0 Maow 8.0 2082
184 Unies 24
Cel syntheses new glucose in response (aluconeogensis - depletion of oxaloacetic acid).
Wormone sensitive lipase (161) helps in conversion of TRE to fatty acids and glycerol
Insulin generally inhibits HEL. Gut in case of diabetes there & excess of HSL + TRe tree —
‘ratty fold ‘iyeerol B oxidation 1
Cone SPS depated KS Tea Cycle thetone body synthesis ‘THs isthe reason for ketoacidosis in
uncontrolled diabetes.
Tests for ketone bodies oor14:17
• RolegrAis PStiecition6notes Purple ring at the junction of a liquids.
Positive in : Peetone and acetoacetate.
4. Gerhardt’s test + Positive in : Aeetoocetate.
5. Betostic: Dipstick test to detect Ketone bodies. Positive in Aoetone and acetoacetate.

4, enaymate assay for BOM butyrate, fredominant Ketone body sythesed in Ketoss
B-on eutyate
} proreutyrate : Reetoacetate = 61
i etone body predominantly synthesized in normal condition +
B-oneutyate:rectoacetate = 111
abertiny std teeta ene FATTY ACID SYNTHESIS
Steps of fatty acid synthesis 00125
Foty acid synthesis occurs in a well fed state under the
influence of insulin.
so Known 0s de novo supthesi/Luynen's spral
Oiscovered by Feodor Luren.
Site + Liver Kidney, brain, lung, lactating mammary gland.
organelle: extramtochondrial fatty acid synthase system found;
Starting substrate : Reety Con.
Source : Pyruvate dehydrogenase (Fed state) (mitochondrial.
Transport of acety) con:
fionBrdes
Trcarboae transporter Citrate. re cate pce (TF feety Con + Ousloacetote a
Blochamisy v4.0 Marow 80 2022 156 Upids 25 Enzyme systems 000658
1. cetyl Co carboxylase system:
Hs a homodimer and is X shaped. ‘ach monomer is divided into 3 unis/domains :
Slocharmiatry - v4.0 + Mawow 6.0 + 2022
Condensation unk feet God rer, Pe Aber reatease of CO tuo AC. oa compounds Sormed of
the ends of each subunit combine to form comers rea 4 > ceoacety Gtwacfrescie ssid
Ha ws Fes o Ea
fey 60) i i Gr By G0 $orms a. eC compound ater release of CO, This compound. hen
undergpes repeat condensation § reduction to gve rise 49 at C acy compound attached to
FA synthase complex.
Releasing unt Thioesterase (deacyjase) enables release of completely
Formed fatty acid from the fatty acd synthase complex.
Cofactor requirement : © NEOPH: IR, ER. Sar
« Gotan corborjase a CE cap
sons derrare” HP (oxicative). mac enzyme. Cytosolic 1COH.
Biochemistry - 14.0 Marow 6.0 + 2022 158 Ups 25
2) Sate lmiting encyme : fety) CoA Carboxyase. Alosteric activator of acetyl Co carboxyose
: Citrate.
Citrate converts the inactive dimer form to an active tetramer.
Alosteric nhbitor of TCA transporter : Long chain fatty acid.
epson iid Gare) Repose
Levers (inactive) Sucagn§ J eprephrne
(@marrowedition6notes
Blochamisty 4.0 Marow 6. 2022 CHOLESTEROL AND BILE ACIDS
Significance of cholesterol Exclusively anmal sterol Cholesterol elimination is difficult.
Chemistry a7 carbon compound (euclopentano perhudro phenanthrene re.
fmphipathic (has both hudrophilc § hydrophobic ends.
Cholesterol synthesis 00445
Site: All nucleated cells Liver, adrenal cortex, tests, ovary intestine. organelle: Cutopiasm §
Se. Starting moterial: Reety CoA. (@marrowedition6notes Steps:
+heetyj con | ve con synthase (aytosoled wre, con
Hime Cof reductase
in endoplasmic reticulum)
• me lng step mevalonate
NADPH NADP
ax Isoprenoid (sopentany unis) Gerany ppi G00) +(sc] (0) Parnes ppix a (300) Sapalene
Bicchamisy 40 Marow 60-2022
160 Las
26
Lanosterol (* cycle compound/{” steroid compound)
l
(10) cholesterol
Fate of cholesterol (a10) Cholesterol
{11
Unabsorbed Sofie acds VHD Steroid hormones
‘excreted (corticosteroids,
through feces sex hormone)
(coprostanal
Regulation of cholesterol synthesis rz Tig reqated patrony
Rate limiting enzyme : Wm Cof reductase. Feedback reguiaton:
Oietory QRRIFFATFEINBTIRRFNG futhesis of enzumes for cholesterol synthesis wih help of
S2e6P (Sterol Regulator lement-ginding Protein).
Feedback inhibition ¢ Cholesterol inhibits HME, CoA reductase.
Hormonal regulation: Insulin § thuoxine Saver HT CoA reductase. elucagon 1
glucocorticoids iho HT Co A recuctace.
Bile acid . wis2r
Starting substrate : Cholesterol Rate limiting enzyme : 1a Hydrojase.
steps ©
Bochamstry v4.0 Marow 60 2022
[ o Si wor | wee 1 1a Hydrongase ero a EP
Chole acid (primary bie acs) Chenodeosychole acid
er
Conjugated with glycine { taurine |
Crolc acid (conjugoted bie acids) Chenodeawyholc acid Gn intestine)
Decongugotn § dehurosgation |
Oeonyeholc acid (secondary ble acid) Lithochalic acid Geast enterchepotic @ ey
‘enterohepatic. creulation Liver
Secreted os bie
Cholic acid and chenodeoxuchoiic acid are primary bile acids ond Formed in the ver.
Bie salt + His he Form in ukich the bie acid exists in the bie.
Bie acid sequestrant Ht sequesters the bile acid and excrete it in Feces and avoids i
enterchepatie circulation. i
So, cholesterol wil be excreted in bie form. Therefore, it is used as a hypocholesterolemic
drug,
Biochemistry + we. + Marow 60+ 2022 162 Upiss
27
LIPOPROTEINS
oeSinton Compound pis complexed ust proteins.
Concept Lipids are hudrophobic.
To be carried in the bood i complexed usith proteins. The proteins ae called
apolpoprotens/ apoprotens.
Major classes of lipoproteins o00320
L chyomieron a LoL Gou-densiy ipoproten)
5 HDL Gigh-density ipoprotend
4. 10L Gntermediate density lipoprotein)
5. LoL (Very low-density ipoprotein)
Stxuctugn oO ipORiBRiRgnoies 00
Core of rac Ses Smee sted
Biochemistry v4.0 Marow 80-2022 _—
Characters of lipoproteins 0005:19
L uo LoL HOU Formed |intestne —|uner | romvict | intestine § from ty ner Lpoproten
cassade pothnoyy woe PL b— cre hiatal Function [carry |earry —fearry ‘| everse |
‘exogereue/_| endogenous | chester to | cholesterol | dietary tha, |The tothe | extrahepatic |
traneport fotnelver | pergheral | sue er | and Srgare | cegere | (See rmaxrum Toensity |
Least ‘
3 Oren | content is | raped |
pa [maxtram Tee Omar or ATENEO apa eiperod A opaigo proten Scotent is maxed fpoipo
poe: |e, poe, Apo proten —unqusto | moc, poe chyjomieron. | po © |Pp00 | feo, [foe poe -
ereyes in HO:
1 Lecithin cholesterol acyttransterase (LAT)
Lecthin + cholesterol @mmphpathie? Leer Liysolecthin + cholesterol ester (hydrophobic)
4. Cholesterol ester transfer protein (CeT®) CceTP tronsfers cholestero cholesterol ester
to other
lipoproteins like IL, LOL ete
in exchange of TR
fet gus
164 Lids 27
Lipoprotein @ (Lp (@)] Simiar to LoL. 0 (2) bound 10 apo 8, by a disulfide bond. #0 (2) a
plasminogen analogue, hich may get ated upon by activator (which converts plasminogen o
plasmin. “This can inhibit clot lysis —> Risk Factor For thrombosis.
Lipoprotein 60 (p63) IF there is a. block in bile flow, the cholesterol will be: accumulated in
the ver.
“This cholesterol ul combine with phospholipid and form Lp 6. Lp is an indicator of
cholestasis.
electrophoretic mobiity of Ipoproteins
Pot of applcation
HH -
Chylomicron metabolism ovens
Blochamisy v4.0. Marow 60+ 2022
Nascent chuomicron contains apolipoprotein 8,, that helps in is assembly. Apo C,§ Apo €
(repository : Hou) gets attatched +o chylomicron forming mature chyjomieron, which on
passing trough capilaries gets acted on by ipoprotein pase: (1,10 present in endothelum LL
converts The fo fatty acd § gers
po ¢, present in mature chyomron is the actator of LL. The subsequently released fatty acid
gets deposited in peripheral issues and The content of chyieron reduces, Sorming remnant
chylomicron.
Apo € acts as ligand For uptake of remnant chyjomieron by ver vi receptor mediated
endocutoss.
LDL and VLDL metabolism 00243
VLDL is Sormed from the ir: & 100 helps in s assembly fo Form nascent LOL. Mature WoL ©
Formed after i acquires Cc and €. C, actotes LL which converts Th to fatty acd
and gerd Fatty acid hen gets deposited t periherol
iccnamatry v4.0 Marow 80 2022
2 eifie 166 Lids 27 ET organs. VLDL with reduced TAG is known as remnant VLoL/ Io. Fate
of remnant WoL © + weepror mediated endocutoss by ligand apo nto Iver. converted to
Lok. (eholesterol/cholesterol ester rich) by the action of endothell/ hepatic pase. 70% of
LoL undergpes receptor mediated endoaytoss into Iver catalysed by ligand apo. 30 of LoL
taken up by extrahepatic tissues that have LoL receptors ise heart or adipose tissues. Inthe
presence of free radicals, LoL. gets oxidised ond is taken up by macrophages which can be a
risk factor for
atheroma. formation. HDL metabolism 00:32:25 vou Croestera frocpholped
w@marrowedition| [a era Jey Wo Per pheral organs
HOL 6 formed $rom liver and intestine.
Lipid content of newly formed HDL consists of amphipathic ips lie cholesterol and
phospholipid which gives it a. discodal structure (axa. discoidal HOLD.

90 8 akiates LEAT hich converts cholesterol cholesterol

ester, a hdrophobi pid.
Vurophotic pds gets nternalsed giving HoL a spherical
Blochamstry v4.0 Marow 60 2022
shape (aka spherical HOL3), This HOL accepts cholesterol rom peripheral organs with help
of certain transporters. Transporters in Ho. metabolism ¢
ATP binding cassette protein A (cA D
a ATP binding cassette protein & (606 D
3. Scavenger receptor 8 (526 Function : Transport cholesterol to WoL from the
peripheral
orgen.
with uptake of cholestero), spherical HDL-3 swells up and Forms WOL~a, hich releases the
cholesterol nto Iver via 526 transporter. The cholesterol then goes for bile acid synthesis.
After release, HDL-a becomes HOL-3. This is Known as HEL.
cycle: Apolipoprotein ovata
Apo A: Activate LEAT,
Fa —— marroweditionénotes + Assemble VORL i Iner.
fet os a ligand for LoL. Apo 6, Assemble chyjomicron. Bpo.C, Inhibit CET
foc, inhibit LL
0 + Ligand For remnant Lipoproten (VoRL § chyomcron)
frgnine rin

0, Late-onset Hznemers assease

0 0 Associated with a human degenerative dsorder.

Lot is dangerous because it has tendency to getdeposited. in extra hepatic issues. Aso,
oxidised LoL can predispose to atheroma formation.
WoL is heart friendly as it ransfers cholesterol from extrahepatic tsues to the ver where i
undergoes bie acid
syntheses.
Biochemistry v4.0 Mamow 60 2022 168 piss 28 =
DYSLIPIDEMIA Oupipdemia Wpertpoprotenemia [Fe — friar ™ . wire wine [Ls
Gupertrigiyceridemia) Gipercholesterciemic) Fredrickson Type | Tupe I Type Tipe Tipe v
‘Type 1 Hyperlipoproteinemia
W//A Familial chyjomicronemia. syndrome. Defect in poprotein pose (pL) apo Ca
(activator of LL) fecurmulation of mature chjomicron and mature WoL.
mature very ow density gid (oF 23 cemoant wou Lipoproten elevated : Chfomieron >? WoL.
Lipid elevated : Triacyjglycerol (TA). Cholestero levels wil be normal. Clinical features. L
Recurrent abdominal pain Hupertrighyceridemi. = kumarankitindial@gmail confancreatitis
> Acute abdomen. 2 Lastescent (miky white) pasa. due to raised tracyghycerd ghycerct 3.
eruptive xanthoma. 4. Fundoscopy Lipemi retinol.
Had<
There i no increased risk of coronary artery disease (CAD).
Slochemistry + wh Mamow 8.02022 28 or elf
Type 2 Hyperlipoproteinemia 001040
L Famial huperchlesterdiemia W/W/R autosomal dominant huperchesterdlemia. type 1
(oH Type D. Ve primary hperlpoprotenema. Defect : mutation of LoL receptor (uptake of
LoL in er and extrahepatic tisues. Lipoprotein elevated : LoL. Upid elevated : Cholesterol.
Triaeyjgjycerai level s normal Clinical features : Presents with premature CAD a Corneal
arcus. a Clear plasma. 4. Tendon xantama/ Tuberous xanthoma. (T/C: ste aches tenor.
a ronTyea W/1/ Samilal defective Apo £100 (706).
Mutation in gene encoding apo B00.
5 ROH Type 3
Gain of function mutation to PCSKO protein PLSK9 binds to LoL receptor — Lusosomes
fecelerated degradation of LOL re
This leads 4o raised LOL and chalesterol levels.
4. Autosomal recessive hupercholesterolemia (12+) mutation in LoL receptor adapter
protein (clears LoL $rom bod.
s. Sitosterolemi.
Sitosterol s a plant sterol. ATP binding cassette : RBCS and RECEE + Intestinal cells :
Retively secrete absorbed plant sterols back to the lumen.
Blochamisy v4.0 Mamow 6.0 2022 170 Lipids 28
• Iniver + excrete plant sterl from hepa: celts into the ble. mutation of ReCaS and fECa®
> Raised celular sterol Decreased transcription of LOL receptor eae iceagas eaised Loe
‘Type 3 Hyperlipoproteinemia o020:59

/n/A Familia! dysbetalipoproteinemia, Remnant removal

disease, Familal broad beta disease. Defect : mutation of apoe : Aets as ligand for uptake of :
+ Remnant chypmicron.
• Remnant Vil. Lipoprotein elevated : Remnant chylomicron and VLDL. Lipid elevated
TAG and cholesterol
Clinical w Palmar xanthoma.
Sight inereased rsh of CAO.
Eriowodition6|
Recent modalities of treatment for
hyperlipoproteinemia 00.2823
Type! : Lipogene Tiparvovee.
ain of function Lpt variant: Type a Familial horozygous hypercholesteroleria) 1. Lomitapide
: inhibits microsomal triglyeeride transter
protein (mre). TTP carries TAG to VLOL and chyfomicron. Results in decreased LOL.
4. Mipomersen : Antisense oligonucleotide therapy,
‘Blochemsty v0 + Marow 6.0 2022 28 one gigremitioe
Hypolipoproteinemia oost.23
Tangier’s visease : Mutation of ABCA gene > Decreased HOL formation, FOCAL transfers
cholesterol from peripheral organs to HOL Clinical features
enlarged greyich tonsi/orange tonsil Peripheral neuropathy,
Abetalipoproteinemi
efect in mT? :
Decreased VLDL and Chyjomicron :
*+ pecreased intermediate density lipoprotein GL) > Decreased LOL.
• Deficiency of fat soluble vitamins @0,¢)).
Clinical Features Diarrhoea. Foire to thre. @marroweditionnotes Neurological
manifestations. Progressive pigmentary retinopathy, Aeanthocytes. eieeding
manifestations.
LEAT deficiency : Cholesterol + Lecithin “FT _, cholesterol ester + Lysolecithin, Types:
complete’ Norum’s disease. Portal: Fish eye disease. Norumis disease # Complete LCAT
defieiency : Raised cholesterol and lecithin. Decreased cholesterol ester and lysolecithin,
Clinical Features Progressive corneal opacification. Progressive end stage renal disease.
Blochamisty + v4.0-Manow 6.0 2022 172 pias 28
Question A 15 year oid boy presented with recurrent episodes of abdominal paints biood
was drawn for investigation and it looks like a cream of tomato soup. One. ood sample Kept
in the reSrigerator over night, o creamy supernatant layer separated.
fosuver + Thi is a case of Tupe | yperfpoprotenemia.
‘Question + & boy presented with the Following features. that is the diagnosis ?
Bo! i Za BE. Gry
Question : Can you correlate ? + Low level of WoL + mononeuritis multiplex
• Mepotosplenomegply
Prsuver + Targer's Diese
Biochemistry v4.0 + Marow 60 2022 LIPASES
reals covalent bond : Ester bond. Trioey eycerol (The) —— elyeerol + 3 Fatty acs. t
3mo
Class of hydrolase.
Hormone Sensitive Lipase (HSL) _ 000319 Location : Adipose tesue. Function : Mydrolyse
TRG stored in adipocytes. ‘uring fasting state : mange afoy con Fastrgstate fctvein: D
Fasting [] @marroweditigh Gergen 2 Prosphoryjated
+e HeL
Insulin > 1 Phosphatase. ® Insulin o--Q
In diabetes, HSL is active. Hormone sensitive ligase
sadn rcotere acd - pat -
Ea ae lana al
29 174 Ups
29
Lipoprotein lipase (LPL) 001453
Anchored to endothelum of copilaries in heart, adipose isu, spleen, renal medulla, aorta,
daghragm, lactating mammary gland.
Anchored to wall by a. 6G > Heparan sulphate.
Injection heparin > LPL dislodged.
fetated by apo CI
Action ¢ Mydrolyse THE: in chyjomicron and LoL in fed state.
Hormone : Insulin increases expression of LPL.
Hormone sensitive lipases vs lipoprotein lipase 002030
Location > Adipocyte copilaries
Acton > ydolse TR In vydrolyse TRG in edipose tesue Chyjomicron, WoL
feton => Fasteg Fed @marroweditionbnotes + Glucagon sulin
100M : MSL will be activated and LPL wil be inhibited.
Hepatic lipase o0zszs
Locaton: Siusodal surface of ner. metabolism of chujomicron remnants and conversion of
oL ton, Endothelial lipase 02630 Action HL, —— wo,
ere Bune
Pre f vow + Poor pated, mest active HoL- sor maximum cesterd fom per pheral orgs
Blochamstry v4.0 Marow 80+ 2022 fie
Intestinal lipase ws
Hydrolgses TAG (dietary) in mtestine.
Fatty acid + eycera
meas 1.10 a ype | Om, which of the Following is correct ? # Actation of LPL ond actwoton of
HL.
8. Rotation of LAL and activation of WSL.
C. Inactiaton of LAL and inackiation of HL.
©. Inactivation of LAL and actnation of HL.
(@marrowedition6notes
Blccnamatry 40 Marow 60 2022 18 ee OO ==: CHEMISTRY OF AMINO ACIDS
Problem based questions - 00028 I Hbs 6" position of beta globin chain, glutamate is
replaced
‘by valine. in deoxygenated state, what kind of mutation leads ‘0 polymerization of Hs,
leading to sichle cell disease?
[Answer : Nonconservative mutation,
Assertion (A): Leucine is present in the interiors oF albumin. Reason (@) : Leucine a non-
polar amino acid
& Rand are true, @ is the correct explanation for A
i. Bande are true but 2 & not the correct explanation
fora ii, Bande are false. iv. Ais true but @ ic fase. @marrowedition6notes
feswer A and 2 are true. Bis the correct explanation for A. Introduction + " i mace *T ey
SORE SBD ONFIOG
carson,
Alpha. osrino acids + ‘frino and carboxy) group are attached 40 alpha. ‘carbon atom. j most
amino acids belong to this group.
Nonralgha. amino acids B olarine, B aminoisobutyrate Y sobutyrate.
Biochemistry «v4.0 - Marrow 6.0 2022 P— cress ao ape Classification of amino acids : pi *
Gedo sd rin homers * Gove mete bt. + Geto miremarenens
Classification of amino acids based on side chain: " " | |
AM, ~C —COOH + by — C —COOH 0) +
° ® wo “ " Coit coon EL Forms. BAB weditionnotes Bond. Peptide can only form hydrogen
bond.
In Jong polypeptide chain, there is free ionizable group a the ends -K4, group at one end
and ~COOH group at the other end.
Characteristic of polypeptide chain & determined by the Voriabde side chan.
fighatic amino acids + Simple amino acid example : ytne(6iy/&) and olanice®o/, + ranched
chain amino acd, example Leucine (ew, isoleucine (s0/D, valine (a/v), (mnemonic - LVL
Fugdrony ooup containing amino acids Serine (ser/S), treonine (The/ 1), yosne (Ty.
Sulprur containing amino acids | Cutene (Cy/0), methionine (met/m.
Biochemistry v4.0 - Marrow 6.0 2022. 178 Protensand 30 Amino acids
fide amino acids Contain COOH apoup in the variable side chain Example Aspartic acid
(Rsp/0), gjutame acd (6/0).
fades © Contain COM, group in the variable side chain ‘example: Rsparagine (Rena,
gutamine (6n/Q).
asic amino acids : Histidine (4/+) > Contains imidazole ring in the side chain.
erginine (rg/2) : Guanidinum group. Lysine (Lus/kD + epsion amino group. wn, Nw
Aromatic amino acids
• Phenjalanine (Phe/B) eenzene ring in the
side chan + Tyrosine (Tyr/) Phenol ring in he side chain. * Trugisphen STA neele ig in the
side chain.
0 Qo
oh doe toe ben
ry Ts
min acid amino acids | Proline (Pro/P) : Pyrrolidine ring ; Aipha. carbon atom i a. part of
the ring.
Bochamisty + 4.0 Marow 60-2022
err 0 on lime
Classification of amino acids based on side chain
characteristics 00:17:50
Interior of a gobular protein contains. non-polar amino acs. (rot sobible in water).
exterior of a gobular proten contains polar amino acids (soluble in water).
Conserve mutation: fn amino ac replaced by another amie ask of sivas characterises by a
missense mulation example One polar aming i seas Pg appter polar amin asd Non-
contersatie mutation: in amin aid replaced by another amino acid of different
characteristics. Bxampie : In
Hos, glutamic acid, a polar amino acid is replaced by valine, a
on-poar amen acid amo acids TN + on polar Jered shee + aromate Crorged uncharged © Sek
+ supe + base Contonieg * simple + imo acd i Saad i
Reid and basic amino acids. + feide : Aspartic acid, glutamic acid * ease: Histidine, Arginine,
Lijsine (Mnemonic : HAL).
Blochamisry v4.0. Marow 60 2022 180 Prot a
ns and
imino acids
30
uncharged amino acids © + Contain hdrowy group © Serine, threonine. + amides :
fsparagine and glutamine. + Sulphur containing amino acid : Cysteine. + Simple amino acid
+ aieine.
most polar amino acid & arginine. Least polar amino acid i glycine.
Non polar amino acids : + ranched chain amino acids (LN). + Aromatic amino acids except
histidine. * Simple amino acid : Marine. + Wino acid: Proline.
Classification of amino acids based on metabolic 002554 @méasdonimetehindote
/ LN
Fetogenic oth ketogenic ~~ Slucogenic
“lew and glucogene. + Alothers ‘ug “ene
0
TTY
“Tp
retogenic, gucogenic and both ketogenic-ehicogenic amino acids. -
‘@lucogenic amino acids provide glucose in the Fasting stage.
Fetogenic amino acids provide Ketone bodies in probnged. starvation.
¥etogenic amino acids : Leucine, lysine. Both Ketogenic and glucogenic amino acids :
Phenolanine, coleucine, tyrosine, ruptophon. ‘hacogenic amino acids : Al others.
Blache v4.0 Marow 60+ 2022 © Cio
Principle glucogenic amino acid is alanine.
Classification of amino acids based on rustritional requrements
Nutritionally essential and nutritionally non-essential amino acids.
Nutritionally essential amino acids : Cannot be synthesized in our body (takes 77 steps to
synthesize).
Cause negative ritrogen balarce i not taken rough det.
example : methionine, Threonine, Tryptophan, valine,
isoleucine, Leucine, Phenalonine, Lysine, Wtcine, arginine.
(semi essentiaD.
(mnemonic : METT Vi Pry + Histdne, arginine).
Nutritionally ron - essential amino acids : Can be synthesized. nour body.
@marroweditionénotes
‘very protein hos an amino acid score based on amount of essential amino acids.
Complete proten Contains all he essential amino acs. Derived amino acids oo3sss
‘Derived from a standard amino acid
Slordard amo acd] verwed amen act con = =
modification | Translatiopal/co-transiational | Post translational cope [ups prone
Classification of derived amino acids + i Seen in proten: i prone and hgrosroine: Formed oy
gboxtien
buy hudrojase enzyme. They are present in calagen.
Via C and alpha Ketogutarate are co factors for
Blochemistry v4.0 + Marow 60+ 2022 a. -_r nyboase enagne. ra ard mE BS SSRI, amis
requked ‘example : Clotting Factors -a/1/9/10, protein ¢, protein s, cotta epoca are cir gsm
ao rte
desmosine ered from kine It is present n elastin.
metry see: Present n skeletal muscle protein (myoseD.
Not seen in protein: + omithine, arginosuccinate, eitruline (Part of urea
• Momoserine, homocigsteine (Sulphur containing amino acd.
sel
• Codon is Ue (stop codon).
• Formed by recoding > Stop codon is converted to coding codon. acid) > Cysteine >
Selenocystene.

• Proteins/enzymes containing selenocysteine Thioredoxin reductase, ghiathione

peroxidase, deodinase, selenoproten P, glycine reductase.

• selenocysteine is oko called as. ar" protein forming amino acd.

Marrowedition6notes rd amino acid.

• "aa proten forming amino acid.
• Codon s ube,

• Formed by recoding.

• Precursor amino acd s lyse.

———
Biochemistry + v4.0 + Marrow 8.0 + 2022 cer 0 life Properties of amino acids 0043.38
Amino acids can © + Absorb uv light. + exhibit somerism. + ext in diferent charged states. +
exhibit bubering capacity,
sor ight
s Amino acids are colorless as they do not absorb visible: gt
• Proteins can be estimated by aksorphon spectroghotometer os amino aids can aborts
UY ght:

• Amino acids absorb 350-390 am of UV light. Maximum. absorption is Sor a80 rm.

• mati soni aids aback 67 I 6 Wo ie congo ring siructure. bap Tryplophan rast, prensa
R9RE tion 6rotes

Shit somerism Amino acids exhibit isomerism due 40 asymmetric carbon

atom. Four valencies are occupied by four diferent groups except glycine. Glycine is
optically inactive and do not exhibit
isomerism.
cine: W | | HC — COM AH, C COOH & —¢ ° e
mirror images ‘Due 4o presence of asyrmetric carbon atom, mirror mages
Biochemistry + 4.0 - Maow 60+ 2022
eceanoy
in structure can be seen. They are Knouun as D and L isomerism.
most amino acids exist in L form as enzymes can act only on L amino acids.
Pacemase is an enzyme that converts 0 form 40 L form. Henee, it can act both on © and L
forms.
D-aspartate and O-serine are D amino acids present in brain
Isoelectric point 0049.26
NM, group can accept a proton and can exist as NH,” “COOH group can liberate a proton and
can exist as COO” 0, there are two ionizable groups.
lonization constant (pH) is a particular point for an ionizable
¥uP- @marrowedition6notes
‘koelectrie point is average of the ionization constants of the ionizable groups present in
the amino acids. isoelectric point is nspa)/a = pl hen the pl of the medium is equal to pl
then the protein is called as 2nitter ion or amphoiytes. n a zuitter ton,
positive charges = negative charges > Neutral
At pH= p's
Thereitinom red Ang

• No mobility in electric Rela.

• cannot attract net water > No shell of hydration around the protein > maximum
preeitability and minimum solubility,

• Least buffering,

Blochemisty + v4.0 « Marow 6: 0 i fie

1 pH < pl > Positive charge (protonated).
Albumin s negatively charged in boo as pH of blood is more han pl of albumin.
Inthe stomach pH less han pl of albumin. Hence albumin carries positive charge in stomach.
editionBnotes
Buffers are solutions tha resist changes in pi. Henderson Hasselbach equation pH = pia + og
base/acid) maximum bufSering capacity is seen pH = Pia. example Imidazole group of
histidine (pra = 65 10 74 = pH of bod).
Blochamisy - v4.0 Marow 60-2022 186 Protensand 31 Amina acids
ET FIBROUS PROTEINS
Structural proteins : Collagen, elastin, keratin, Sbrilin, laminin. Collagen — 0207
most abundant Sbrous protein present in extraceluiar matrix most abundant protein nthe
bocky. Highest density in: Cornea. > skin Structure of collagen: Triple heli: 3 Poluproine a
chain. Sige poly proline a chain > Glycne-x- repeat. ‘every 3 amino acid > glycine. % 4 = tayo
proine/ dro lysine ach @ chain made of 1000 amino acids mosabundant amino agidié ghee.
23% of @ chain is alyone. Recurring amino acid. each & chain twisted in let handed
direction. @ chains together tusted in ight handed drecton.
Quarter staggered arrangement +
iat Si oe fe
aka from wales RYE cross tows
v0
Blochemisiny- v4.0 + Marow 80-2022 le
[— + ydrougation of proline and sine residue
| vitamin, a vetogutarate 4 proly/ lysy hudrowjase Gronooxygenase) eee elgosujation of
hydromy sine. Intra. chain and inter chain disulfide bond formation. Formation o triple hel
— eolg apparatus frocalagen packed into secretory vesicle. Transported to extracelular
matric extracelulas events Cleavage of N and C termindi{eysepsads Rssembly of collagen
Sri into quarter staggerred. arcargement.
Formation of covalent crosslinks.
Types of collagen
Ye | sk omecte tet Sit GGRIEMon Grotes
bcm
Ueda comma tes sas i sd
vascular
[| mor corpora sms carn bg |r rts
fchoring Serie.
fom pa
| perp carige. | x | Tesues contain cotagen | xi | Tasues containing colagent.
xv | mony Tesues. | man Teen i
Biochemistry +40 + Marow 60-2022 mo major collagen present in bone : Type | (0%). rp
cade meat Sa pa soca Type | (®0%. major collagen present in cartiaae : Tune I (40-50%).
agen pa AA SABA BEGHRS Te major collagen present in Rorta.: Tupe | § Tupe Il (20-40%
each) and most abundant collagen : Tupe 1. Eagan aed ig Co ry A At i
m/e + Tupe 1 colagen. Type of collagen and associated disorders onaze Tupe of ene or
Dense agen | erayre Type Coan | Osteogenesis perfect, couna ehers-oanios supdrome (Tupe
vi ec) pen coun Crondodppasas ostecartrte Tp cot ehiers-parios syndrome (Tupe Iv £06)
(mos serie) open © TIEIRIOTE 2S prome Gece ot come autosomal and x-teved ferme)
Typevond | cous, Chasscal 05 Tipe casa, coun Type co Tupermobie £05 (Type 1 £05)
Terasen xB (The) Type can epdermongss bulosa, Agtrophe Type x coon Schmid
metaphyseal i crondrodsplasa uy py erers-oarios ngoyase | house | (ype vio) huphoscolote
£05 Scurvy i trent some i Wporatass | Ghgah utensyel rosnihed (aio cated os |
vermatosparoxs tupe: sogen 752) upg Lupefoccase | merwes dsease GTPe) oxidase quires
Cu)
Biochemistry - v4.0 Marrow 8.0 + 2022 St roe igure
Vilefranche classification of £05 +
Eras or 1 iypermobity “Type eolagen, tenascin x a classical Types and vealagen a vaseuar
“Types ealagen “a raproscoions yy rpronyjoce 5 fthrochaase Type‘ eotagen foam
metalopepiaace wih 6 vermatorparans Ahrombotconda ue mot Goeertt Sar y Elastin -
00:95:21 elastic recod. Lung, large arteral blood vessel, elastic ligaments. - eotagen | eastin
| 1 Types many ony! a. Triple Helix @marrowetiition6notes— a ype +] - 4. Presence of
hydro ysine + -
fit | cesresne + -
Keratin a a _ 00:38:54
Froten present in the hat, ras and outer layer of skin, i
Alpha heli eoesIned by dsiphide bond,
Rich in cysteine. Warder the keratin, more is the disulphide bond.
‘Biochemistry 0» Marow 6.0 = 2022 190 Proteins and Amino acids
31
Fibrillin-1 ovav1s
Large giycoprotein. Structural component of mieroforis. Scaffolds for deposition of elastin
Mutation i gene For fbrilin- leads 4o marfan's syndrome. fo: Reromere dysplasia.
Salsopbuzeh dueplasio:
Congenital contractural arachnodacty :
mutation in the gene of Florin a (chr 51
This is important in deposition of microRbris.
early in the development.
Clinical features : Contractures, frachnodactul,
©olchostenomela.
Classical epidermalysis bullosa : mutation in heratn-s.
meas: which i¢ the following not a Rorous protein ? # collagens marroweditionénotes
6 Blast.
C heratin.
© Myoglobin.
‘Desmosine cross ink is found in A elastin.
©. reratin,
¢. Collagen.
©. 5ik Sbron.
& Laminin
which of the following regarding keratin false ? Present in hair, nail and outer layer of skin.
©.2ich in cysteine. Gocsbassaasdedo: C. beta. plated structure.
©. The more the disulphide bond in the keratin harder the structure.
Alpha. hel cross linked by disulphide bond.
7
Blochamisry 140+ Marow 80+ 2022 ye
The disorder associated with deletion of elastin gene A marfan syndrome. &. Congental
contractural arachnodactyjyy. cep. © wilam Beuren syndrome. © epidermis bulosa. which
of the following statement regarding streuture of collagen is/are false 7 A Forms unicue
triple helo er camel Sivas pattern. each polypeptide chain s twised into right handed
polproine heloc ©. Sach polypeptide 1s approximately 1000 amo acids. © Tupe 110
glycosidic inkage present. Recurring amino acid present in collagen & A Ggine. ©. Proline.
CHa pon @marrowedtion6notes © Hydronusie. match the Following ‘epdermolys bullosa :
Type VI collagen. ports syndrome s Tupe I collagen. menkes disease : Lusyl oxidase. Sound
(Op rocks Schmid metaphyseal chondodusplasia.: Tupe X calagen.
OR: Type I collagen. The gene defect in olport syndrome &
Acouan. cco 4.
&.cou sa. o.coL
The tupe of colagen Sound in anchoring foris ?
aa Tan i amen oTypewn i
Blochamaty - 40 Marow 60 2022
es 2 -— GENERAL AMINO ACID METABOLISM
Deamination & decarboxylation o0or04 deamination" decarboaaton
©=C- COOH ©] KH, = C= COON | AM, ~ CH,
wa co, |
® ° eo @- veto acids Handing of amino group :
I. Transaminotion.
a. Transport of ammonia.
3. Oxidative deamination.
Transamination = us
Deevtion : TransFer of amino group from one amino acid 10 a erBSErO OTTO par oF amino
tes
@nEFHAPAES AS Takes place n ll organs/cells. Ammonia. s not released freely Seversicle.
Coenayme : ALP organelie : eyoplasm
SePT/AT (lance Amnotrarsterase)
Rance
portale retogutarate erga |adicbe |
Gitamate Onaloasetate” automate (posh (ron specite)
Transamination is specific for one pai of the substrate but fot specific for the other pair.
@ amino apoup of any @ amino acid is getting concentrated as gutamate (only amino acid
to undergo oxidative deamination). i ee pecs TG FR ARR ls RT ctirsastan
@retoacd a ketogutorate Shtamate [5 €
a amen ei h _ 7
Biosuthess of non-essential amino acids © @ Ketogutarate — Gtamate Pyate — fanine
Oraloacetote — fspartate
The non amino group that undergoes ransamination - 8 amino group of ornithine. ‘enzyme
+ & Omithine aminotransferase. oeSiiency urate atrophy of retina § choroid, Treatment :
Restrict omithine § arginine (source of ornithine). ves cazenzyme ER ion 60 es
Sources of ammoni:
02005
amino group amino acid (getting concentrated as
Transport of ammonia from most organs (including brain) 02033
cri ipa as gas Other sources : Release NH, (tox).
Agha. amino acid > Glutamaed. sistamine ) other sources (3) __ Spetese fre sutamine
Blochamisy v4.0 Marow 60 2022 — 194 Priensand 32 rn wT lutamine is a transport Sorm
of ammania, from mosk organs (including bra). The reaction Hares place in mitochondria.
Frst ine trapping of ammonia. Ligase reaction. Toxicity of aero o brain tan ei — st
permeate)
(excess) | eoodbran varer
ane ——— > automate ———autamne 1
tomes | po
co, 3 = wo 4 daradegeton tf acren | [
neurotransmitter) rence pen cores
edema.
| @marroweditionénotes 4 ATP producton
are doglten
Transport of ammonia from muscle 03001 @- amino acid ara sl ir pe
oho
In the muscle, glutamate is converted back info a—retogutarate and pyruvate takes up
amino group and becomes alanine. Alanine is the transport form of ammonia in the muscle.
Foal destination Liver (urea suthesis can only take place the ned. Gitamate ara st nee rat
Pyruvate
Bocnamstry v4.0 Marow 60 2622 — 32 Gon
Glutamate metabolism 0x17
hatamine (most organs) § alanine (usc) are concentrated as glutamate.
‘Glutamate undergoes oxidative deamination in the Iver. Oxidative deamination
A, released Sreeh) : enters urea cule. SH can use either NAD and NADP* as coenzyme Forms
KROH and NADPH respectively,
GOH oxidative deamination takes place in the liver § Winey,
Reversitie reaction.
Organelle : mitochondria.
Allosteric activator : @fharroweditionnotes Allosteric inhibitor : ATP/&GTP/NADH.
Trans-deamination : Transamination (ecurs in all cell) is -
coupled with oxidative deamination (occurs in the Iver).
Blocnamatry 40 Mamow 6.0. 2022 196 Proisnssns Amino 33
UREA CYCLE AND DISORDERS
Introduction Co mew
50 Known as: Freb's Henseleit cule (scientiet’s name). Omithine cycle ornithine Is
regenerate). Urea Bicycle (inked to another cule).
reeks bee
As OA Ww, co, fspartate
Compounds consumed in the Urea cule ki, (ammenid, CO, and, Aspartate. Site sLiver
(exclusively). organelle : Cutoplasm and mitochondria. Other pothusas taking place in both
cytoplasm § mitochondria. Hue Pathisay
• Heme synthesis

• urea cycle

‘huconeogeness
• Puymidice synthesis
5 fate ke sap fees sfc arts tits sypthetase
Blochamistn 40 Maou 60-2022

oa 3 Disordef Reactions of urea cycle 000735 co, vam,

Carbamoy po, | a ATP wi - Reety Glutamate (Ac)
sytvetaser |( a~eo,
Crs | Manor
carbamay PO, 8)
Ornithine Trans
tor . i omithine Citvuline | “A Chali + spartatd ire + ornme | transporter, 4 Ormitife, AP @~
po) urea” Arginine, Arginine AMP. A, - CO Am, succinate (6) dition TeReyse wgease |) Bont
Crugroase) ™ frginosuccinate Lpse PS 1+ Rate lmiting step. Uses a high energy PO, ocety
glutamate (ie) © the obligate allosteric actuator (positive) of he frst step. energetes
CPs-1: 3°60, a ATP > 4 RO) Frgninosuccinate synthetase : a°PO, ATP > 1 AVE)
• Total 460,
• 38TPs used directly
• 4 ATP equialent used. ceguioton i etary increased protein fake induces urea § cycle.
enzymes.
NRG is a postive allosteric regulator of CPS-I. Compartmentation: partially in mitochondria.
§ party in he cytoplasm.
Biochemistry v4.0 Marow 60 2022 198 Prensa Amie 33
Urea Cycle Disorders oo 02040
co, +n, ps1 Hyperammonemia.
md frgnosuctinate gnnema. arginine © SR J grote Lynas acdura (@marrowedition6rotes
Hyperammonaemia & plasma glutamine 002422
Relationship between hyperammonaemia. and plasma.
Jutomine : 11104, ctamate 4 11 cutamine Types: — Tupe-t Type-a Cw ow, Plasma Giutamne
> Plasma Glutamine
7 Type-a ruperammonaemia. i eSecte enzyme : OTC. ures ncreased.
Biochemistry v4.0 Marow 60+ 2022
19 = Soa
Characteristic features + + clined partially dominant (only males affected. + increase orotic
acid secretion + most common urea cysle disorder (40).
Trichorhexis nodosa. +
Tuttedhor —_ @marrowedition6notes Seen in argninosuccine aeiduria.
HHH syndrome + Hyperammonaemia hyperornthinaemia, omocitrulinuria syndrome. cue
to defective ornithine transporter «
• ornithine accumulates > Hyperornithinaemia
No ornithine : CP had nothing to combine with > hyperammonaemia
• CP combines with lysine : homocitruline > homocitrullinuria,
Arginemia. Least hyperammonaemia.: Arginase is the last enzyme. rginase has a isoforms.
Progressive spastic dplegia { sessoring of gait.
‘Blochemisty* v4.0 = Marow 6.0 2022 200 Proteins and Amino 33
= -_— Clinical features & investigations of urea cycle disorder 0:33:40
Cleat features + 1. encephalopathy a Respiratory akaloss + a4, > Hperventiation
Tachypnoea. > C0, washout. a Hyperammonaemia. 4. Nebases prose warren difSiculty,
lethargy, vomiting, Faure 10 thrive, convulsion. Tochyproeo. Investigation elood pi evel
—— — Low pi Cacidos) vigh pH akalos) + + organic asiduria Urea cule disorder i
@marrowedifbR HERES: aminoacid levation
1 gnosuccinte fs acura OOM acd clevaten 1 citvutine : Cruinemia.
FriTpe-ava 1 orn ve some ome 1 seg egnema.
Treatment wana
First-ioé treatment is argrine © + essential amino acd + Provde omithine + positive
reguintor of ne synthase > increase NAG eps Contraindication : Do nok give i arginase
defect. Second ine : Resfation therapy + Sodum benzoate and phenglbutyrate used.
Biochemistry 40 Manow 60 2022
2 Sat
OF NH, Pheny) acetyl glutamine L
ecreted n urna sodunberzsde —__— ephe + “eens
conzoy aye ‘question: The gen table iS the [B-ropanwe o> ewe eroene
suspecting urea. cycle disorder. Identify the enzyme involved, in the correct sequence.
Infant] urine orotate | ood citruline | elood | Glood NM, argeine 1 uewe [vow tow | wigh a |-
| wgfad marro vfeaition Gmgfes | C1000 um) a [- ~ igh | moderately rea + [igh von vow [gh
Options : a ces C. Pegininosuccinate synthetase ore ©. Agnase
Prswer! 148, AC, 3-0, 4-8. Question & month old boy admitted with aire +o thrive with high
@lutamine 4 uracil in urine, hypoglycemia, high blood ammonia. Treatment given for a
months. At 8 months again admitted for failure to thrive to gain weight. Gastric tube
Seeding was not tolerated. Child became comatose, Parental dextrose given. Child
recovered from coma. within, 436 hrs. what i the enzyme detect ?
A ces, ©. Ornithine transcarbamoyiase.
C. Prginase. © Arginosuccinate synthetase.
Biochemistry + v4.0 = Marow 6.02022 202 Proteinsand = 34.
rae = AROMATIC AMINO ACIDS,
oo007
on, 4 O removed tayrog + group OD on 4
Prenat rng Essential amino acid (ae) Non essential amino acids oth oth Ketogenic §
glucogeric
vo pani ue ato TION GBSE Ron polar among non polar aromatic amino acid,
Phenylalanine & tyrosine: Metabolism 00:06:25
Phenyolanine ——> Tyosine
meni ioe en el a
Tetrahydro- Gyro curser bape
ciafctaptern redctase noe bP
Blochomisy + v4.0 Marow 60-2082 34. Arom eds
Properties of this reaction + Wrreversible reaction,
• Phenyalanine hydroxyiase i monooxygenase. Concepts of Phenylalanine hydroxyjase
(Prt)
1. Phenyjalanine > Tyrosine
(6 an essential amino acid, Typosine ¢ a non essential amino acid.
. Phenyjalanine doesn't have any function in the body per se. ik converts into tyrosine and
enters into a metabolic
petranys
Catabolic fate of phenylalanine and tyrosine __00:1406,
youve troneamnase “emananedto masuewedition6notes Prenyolanice”= Tyotne —— PUES
hong pywiate hin Diem (ove) « cok,
i
‘Biochemistry + v4.0 - Marow 6.0 = 2022 oes -—r - Csi prenyretura fect hereto hudroxjase
(PAD. + Non classic phenylketonuria. type I § 1 + Defect in dproopiernodutase + Non classic
phenylketonuria. type Iv § V + Defect in formation of tetrahydrobiopterin. Akaptonuria.:
Defect in homogentisate oxidase. Tipe nema. cote ene on gcse Type I fyrosinemia.: Defect in
tyrosine transaminase. Toe goin. geet 67 poo
Phenylketonuria 02254 pr Phenylalanine ——>¢—— Tyrosine. Phenyjalarine enters into
alternate metabolic pathos. Trancamination Reduction Prenyalanine > Phengpuruate
——— Phen lactate
@ erode
rclutamioe | Cory
Dramine Pens acetate causes must body cur CC ryene [ l ) mein catesamnes Thugs
us eyes, blonde Neurological deficits hai; Sar skin. Agitation, hyperactivity Some dietary
uyosine mental retardation converted to melanin.
uy
Causes of neurological manfestations |
Inthe brain : Phenylalanine, tuosine, tryptophan passes. Hough the blood brain barr er
through a transporter:
Blochemisny v4.0 Mamow 80. 2022 In phenyketonuria, phenyalanne saturates all
fransporters.
1 Phenyolanie, | tyrosine, § tryptophan. + Phenalanine cannot do any function as it i unable
to convert to tyrosine. + Phenylalanine also prevents dietary tyrosine 4o enter trvougih he
transporter.
I Tyrosine + | eurotransmitters, | catecholamines Uruptopran + | serotonin
CrrcarveTUe YT THANES intractable vomiting. misdiagnosed as congenital hupertrophic
Pyjoric stenosis.
Laboratory diagnosis and treatment of phenylketonuria 0033.34
Guthrie's test (acterial inhibition tos): ests stl ers preplon tr gon. # phenjolarine precedt al
seen. In neonates, heel prick sample taken.
a. Ferric chioride urine test: 1 ml of Rerric chloride added to Urine sample: Blue: green
colour indicates a posite fest Transient reaction.
3. &h0d phenolanine Normal a- 6 mg/dl 30 mg/d: bad prognosis.
4. enzyme studies
5. Genetic mutation studies. a Tandem mass spectrometry : eld standard screening
method for all metabole disorders.
Treatment + Restrict prenyalonine (cassava based died.
Bochemisy v4.0 + Marow 6.0 2022 — 206 Prownsand 34
• Concentrate of large neutral amino acid (ARR,

• Sapropterin dihydrochloride (suvan) : Synthetic 84,

• enzyme replacement therapy under tril: Phenyjalanine ammonia lyase. Alkaptonuria

smooeateate - > Benzogunone X vomogentsate Occase. | potymere Maly acetic acid
(mee) Akaptone bodies

AKaptone bodies accumiote in nose, pina. sclera.’ Back spots.

Homagentisate excreted in urine, gets oxsed and
resus in blackish discopration.
In infants, black reddish discoloration of diaper.
Increased discoloration on washing with soap : Akalnizator ra SH RBnotes
No mental retardaten.
Cirical features : Back, pain, black spots/ pigmentation in
middie age.
ochvonoss ¢ Akaptone bodies accumulate in intervertebral
des.
‘Garrod Tetrad + First inborn error of metabolism studied by Archibald Garrod.
• cystiuria
A - Akaptonuria
A - Abnism
© - Pentosuria
Laboratory diagnosis Rkalicisation of urine : 1 darkening of urine. Ferric chloride test:
Positive.
Rao, test Positive
x rou spine Pasrot beak appearance.
Biochemie v4.0 Marrow 60-2022 or * iden NHinone/NTBC + inhibits PHPP hudroxase and
reduces
homogentisate.
Segawa sucrome + Autosomal dominant.
Clinical features + Dystonia, with durnal variation. Females > males.
GTP cucohudrlase enzyme defective.
44, ut bod phenutanine levels normal.
Tyrosinemia ws12s
Tupe 1 tyosinemia/ Hepatorenal/ Merediary tyrosine. #/i/a tgosiosie.
most common type.
Hepatic and renal aiure.
enzume defect ; Fumaryjacetoacetate hudrolase.
Aecumulation of succing) YB aToweditionsnotes
Liver Faure — MAT enzyme affected accumulation of methionine. (boied cabbage odour)
Resemble porphyria. Treatment : Nitin.
Type I yyosinemia : Wi Oculocutaneous tyosinema/Riner hanhart syndrome.
enayme deficiency : Tyrosine transaminase. Cirical Features Corneal ulcer;
Cutaneous non pruritic hyper Keratotic plague.
Tupe I 4yosinemia/ Neonatal fyrosinemia : Least common $ [—— i mutation of PHP
hudrosfase (partially active)
Gives summing pool odour.
Blochamisty - v4.0 Marow 6.0. 2022 PO ses -—m
Anabolic fate of tyrosine ses?
I Catecholamines Synthesized in the chromatin cells.
adrenal - 80% adrenaline extra adrenal: Nor-epinephrine. Rsv Srliiog ob sumpathets greg:
Tyosioe | Tyosne hudoajase Diy phenylalanine (COPD | copa decarbojase, Pp opamine - kt
catechoiamine | oopomine betaowdase Nor epinephrine N-methyl S-adenosymethionine
#ransferase C <-aderasghramssgtone (@=pieeghrmsiition notes
S-adenosyjmethiorine derived from methionine.
oopamine Nor eprephrne epinephrine | Catechol-0- methuftransterase |
Homo vonillc acid Nor mefanephrine Metonephine monoamio Occase
Vonilyimandeli acid (ed a mein Suthesized in melanosomes in melanocytes (deeper augers
of epidermis).
Biochemistry + v4.0 Marow 6.0 - 2022 4 Tyosne || Tuposinase (requries cu)
Obugdroy phencalanoe (00%)
8 Thyme) Synthesized in thuroglobulin in thyroid Folices. Has I5 fyosine residues. 1
iodinated : MIT - monoiodo hyronine IT - Diodo thyyronine.
Pheochromocytoma oros0r
Tumour of adrenal medulla, excess catecholamines.
2a sig Qmayovediionsnoles
agnosis: a4 rr urinary examination for
• vaniiymondeic acid

• Fractonated metanephrines (highly sensitive) In plasma, catecholamines and free

metanephrines.
Question patient has a “pil rong! treme, “caguheel” rigidity, bradukinesis, speech
difSiculties, and a shufSiing gait. The chemical that i lacking in this sydrome & a derivative
of which of the Following amino acids ?
a Aanine.
8. serine.
C Tyrosine.
©. Tryptophan.
Biochamisty- v4.0 - Marow 60 +2022 210 Proteins and Amina acids
34
Question A.4 month oid infant normal at birth, developed. Sudden tustching movement She
noticed a mousy body odour inher wet diaper. She was taken to a pediatrician, sample
taken from heel pricks was send For uthries bacterial hibition test. QD what is the probabie
diagnosis ? Phenyietonuria (PKL). Qa hat is the reason for mousy body odour ? Phenyl
acetate. 9 which is the amino acid elevated here ? Pheny alanine. @) what is the bacteria.
employed for this test ? Bacilus suis.
‘Question what is the rationale of using this x in PAU? os 0 henyhetonur, prenjolonee I
saturates all transporters, hereby not
allowing tyrosine § tryptophan to enter into the brain. LARA has high amount of
@ rrowediti ears mpl exile henlonce. A —4 mtn: 55 ot by recat a og, bre
Sar skin, eczematous rash, unusual mousy odour Forced to at fruits and other special det.
ensuver : Thi s a case of Pheny Ketonuria.
Blochamisy v4.0 Marow 60 2022
34 Aromatic nn lr
Question 4 year od gr presented to Pediatric OPD with complains of sudden weakness of
both lower lms especially in the evening, but improves. in the morning, On investigation,
Tetrahydrobepterin level is reduced, but Peng flasine level normal what s your diagnoses?
Answer + Segawa. syndrome.
‘Question Identify this disorder associated with catabolic
potty of pheny alanine ?
rrowedition6no esi + Type I Tugosinema Gujperseratone paces, Corneal uleer). ka.
Oculbcustaneous tyosinemia/Richner hashart syndrome.
Question : A & month year oid boy presented with severe metabolic acidos, ketosis, Sailure
to thrive and unusual odour of swimming pool as he weaning started.
Urine Tandem mass Spectrometry - Houkinsin,
frsier : Haukinsiouria.
212 protons ana Amino acids Sh
Blonde har, far skin blue Miky white har, more eyes. depenented/miy white This s a case of
PALL skin, red eyes. This 5 a
case of albinism.
melanie synthesis s melanin synthess is offected. affected.
Tyrosine is 3 WERion Go BE Srose enzyme self is melanine. deficient.
fut dietary tyrosine canbe Even dietary tyrosine converted to melanin. present, con not be
converted to melanin.
GLYCINE AND SERINE
Glycine 023
en Teonne 3 Shine Sere syease

serve
Catabold fnabolc——¢,C of purine > createine ob — sere signe [> atathone l ceaioge Pyiate
sytem. (Queogenic AD
W Chemistry ~¢— egos /optealiyacte " carbon atom
Non essential amino acid.
Purely glicogenic amino acid. Polar amino acid. Serle amino acid. Glycine : Pathway ous
Swen (LP, ole acid Serve > Giysne SHIT = Serie hudrony
methwjene THER (B Carbon) methy transferase 4
Primary donor of I carbon group
Biochemistry v4.0. Marow 60-2022
35 N— 214 Prownsand 35
fe
Gionjate larine aminoirancferase (PL) agate © - seine
ZN
/
Woh Ronis apajatec ~elutamate
apres aneguiante
Threonine akdolase Threonine ~~ sgene
Glycine Cleavage Stem (60) rultenzyme complex. 4 - rote covelentsy loved 0 3 nciyres). D
@lyeine dehydrogenase : Cuts the Coo. Aminomethy transferase : Separate methyl group. i
Dhydroipoamide dehydrogenase.
@mrrowedition6notes
Bog wok AED co,
"i
HTT ven MN, methyeneTiFR Glycine : Anabolic fate & function 0:19 crestiva - — — —
ehyons argne ! wind i be as dng © I “andeo i ee" Fortes methanne i ner Creatine ¢———
Creation phosphate we me iE 35 Gheine sone Siete sm: S-Adenosy| methionne. ‘Sl: S-
Adenosy homocysteine. Creatine PO,: immediate replenisher of ATP in muscle
Gest 3 4 seconds of exercise).
igh energy compound.
a/k/a. Phosphagen.
Heme Suceiny| CoA + Glycine > —> —> —> Heme
‘@lutathione #
CCN, of purine,
Neurotransmitter
Conjugating agents Bile acids. @enzoyl Co A: Hippuric acid.
most abundant/recurring amine acid in collagen
Induces bends in a structure of proteins.
Glycine : Clinical correlation:o wocitionénaies 902808 + reporter, ph rape“ deen pape Le
ftnee fretrorterse + vae,deteony —| | mpea—asfestneiperte matewg =
deyprogenase/eypaysto + methoufurane —| reductase + ejere eat — L____i neg cepts
eceuriaion + enteric — 4 raperoin aura
VF deflect occurs in GCS : Non-Ketote hyperglyeinemia.
serine oars most common site of phosphorylation. i ¥ + Polos, uncharged amino acid Nh, —
G— COOH Non - essential amino acie CA,» Purely alucogenie amino acid
Biochemistry v4.0 + Marrow 6.0 2022 now [a
Can be synthesized from: Giyene <> Serine, 3 Prosphogycerate (369),
metabolic functions :
Primary donor of 1 carbon group. serine» alycine
maton
custene synthesis.
Phosphatidyl sere.
sphingosine (serine + palmitou) Co ©.
Selenoeystene precursor.
serine PP ethanciamine “eo, i Siycoprotens Chole (Trimethy ethandamine) i [] O-inwed
gugoproteins Getaine (Trimethujapycie)
(@marrowedition6notes Most commen site for phosphoryaton (covalent
modification of enzymes).
which & the amino acid involved in heme synthesis ? igre.
a. State true/false ? ‘Amino acid can conjugate compound in xenobiotic reaction : T.
3. Why oxalate stones in urine in vkamn 86 deficiency ? Because of deficiency of alanine
ghyouate aminotransferase.
4h the amin sid completely ecsrporsed fo puree rng? age
5 Compound derived from glycine are all except? A. Creatine.
Biochemistry 4.0 Marow 6.0. 2022 ©. aitathione. < purice. ©. Pyrimidine.
State true/Saise :
The follwing statements are regarding hyperoxaluria.
A Vitamin eb deficiency can couse # : True.
&. Primary type is due to defect i alanine gyoxsjate amino transherase defect : True.
€. Vitamin C deficiency a couse : False.
©. icerate dehydrogenase defect can cause i True. £1416 peroxisomal targeting defect :
True.
© rch th Soousng gre Sipjote.
Non Ketohc hperghjcemia: ine essa system Huperosairia: eee gol
Serine 1 C metabolism.
(@marrowedition6notes
Blccnamry 40 Marow 60 2022
35 Geno.
mmm SULPHUR CONTAINING AMINO
ACIDS Introduction 0325 + cystene + methone W w loon SS oh a su Gu, 7 Thoether 5 ]
Levage on Sulfrugdny (-SH/Aricalechal/ Non-polar fA. Hol group. oes nok respond fo Non-
essentiol amino acd (sulphur test Purely gucogenic. 57 Polos SYR Ae ditionenotes Pre
Juesgerie: ‘Methionine : Metabolism 00:08:20
• methionine Not a meth donor
• 5-Adenosu methionine (5a) + Prince meth donor
methionine L methionine Adenosy TransFerase (VT)
Transterase J vomocustene Ga) Povo = |. 7 Homocysteine | Adenosy i Homocysteine +
methionine Adenosy Transferase (mam):
mAT- | => Liver. MAT- Il > Extra hepatic Tissue. AT- > Liver.
Blochemisy v4.0 + Marow 6.0 + 2022 ‘Sulphur
Se
• significance of 5- Adenosy] methionine (Sam) + Transmethation Reaction. Polyamine
Synthesis. oA methyation.
Transmethylation reaction & polyamine
synthesis 01356
| msepor | metyoted pocket cdo acetate [creatine
or pocptree |cpmepteive
epnephrine metanephrine
ethanciamine Choline Grimethylethanamine)
camosine Anserne. feety serotonin | melatonin a. polyamines © + Organic compound with >
amino group + Positvely charged © interact with neget yh hRSOR MBnGnotes Regis gene
exjresson: + synthesis of polyamines : Polyamines are derived from Ornithine and
methionine ormithe.
cate Lig _, bo enne R18) | ornithine decorbouase (rote imitng step) Futrescine
| Decart a fyi se rin
|" vecarkomiated L104, 5m + Sm «= methionine spermine
• Lysine on decarboxjotion : Cadaverine

• Ornithine on decarboajation : Pulrescine

• Other polyamines spermine § spermidine (Precursors : Ornithine § methionine).

Blochemisry v4.0 + Marow 60-2022

36
Amino. 220 Potonsand 36
ms am
Fate of homocysteine 02434
1. Regeneration of methionine
• THEA: Tetra vydro ole fed

• there is deficiency of 8, , | Free THR. A functional deficiency noun as Folate trop/ THF
starvation ‘ ‘Defect in DNR synthesis [) megpiiost anaemia. @marrouifdTesest nine
reas ¥ macrocutes peripheral smear
• 1 there is deficiency of &, /folc fed,

o,/f Homocysteine 4 methionine

oeticencyy of 8, / Fa ——>1 fomocystene |
26k Sato for thromeesis 4 (cao § cv) Coronary Artery Disease. - Cerebrovascular fecidents.
+ Deficiency of 8, and Folc Acid (7): + vomocusteine i goods Momocystine (2 homocysteine
joined by a disulphide ond) excreted in wine. a. Gucogene fate © methionine
Bochamstr v4.0 Marow 60+ 2022 ne
Homocysteine. sere (Coenzyme) PLP | Custathionine f Synthase. cystathionine reaston
(Coenzyme) Pp | Cuystothionase
Homoserine + cysteine.
Proponyl CoA
Suceinyl Co A
TTR
1 vomacistios 0%. roweditionéotes Biochemical disorders of Sulphur cont inoacids ds
1. Cathouse sundrome/Smth strong disease ©
• Defect : methionine transporter (intestine)
a Primary hypermethioninemia : + Defect: methionine fdencsy Tronsferase (TAT) +
Characteristic feature : Boiled cabbage odour
3. Classic homocystinuria
• Defect : Cystathionine f synthase
4. Cytathionnuria: «Defect : Cystathionase. ; 5 Non-classic homocystinuria: §
• Defect :N, methyl THFA and meth ©,
Biochemistry - 4.0 Marow 60-2022 222 Potonsand 36
Classic homocystinuria : Defect and features 00.4225
• Autosomal Recessive (30)
• Bochemical defect : Defective cystathionine f synthase. methionine t Homocysteine
Cutothonioe | serie sythase | “PLP (co-enzyme) Custothionine i
Homoserine + Cysteine
Tromacusteie n shod romocigstae o ure. 1 | cystene suthess. methionine level: ormal *
Clinical features ily SYRIA 10 Loc Later, Developmental Delay, fk 3 ys of age: § Vision,
Progressive myopia, Quivering ir (relodonesi), On Examination: etopa lente. (Lens +
Docated medially and dosrisords) + Skeletal deformities. Grachnodactuy, Pectus. carinatum
Pectus excavotur). + Severe mental retardation. + Thromboembolism. + Genus
valgum/Narum + Coa vara ba
• Pes cams,
• vigh arched polate

* EE me
Classic Homocystinuria - management o0s212 — Ce i ppaiiaiomclcine tis mii. pre pai ire tvs
elias ohm aes Cysteine, cystine. p—— died + igh dose of vitamin 8 Reason vitamin & (PLE)
> Coenzyme of cystathionine Bytnase. + Restriction of methionine with cystene
supplementation. RT
• eetaine supplementabionaroweditionBrotes, Reason: Trimety glycine (Betaine) >
Remethijaton of homocysteine. vomocstene O° methane. octane
• Administration of Vitamin C > Improve endofalil function.
Non - classic homocystinuri 05550
• efect Defect in formation of N, methyj THEA Defect in formation of methyl cobalamine
metrujene Tetraudro Folate (NTH)
methonce | Fre ceducatose i PGS Sra i se, Sm
Homocysteine
l
Biochemistry v4.0 Marow 60 2022 24 poems 36 ww -—m
Cuptathonne |
Cutene + homoserine * Level of methionine § |
cystene : Normal Homocystinuria - Comparison ooseun [Feature classic metry | mre
Pomceytiouria | cobalomine | reductase
foe defect | doboiency serocgtenena - v - methonne evel Normal ' 4 boot Cuptene evel n 1
Normal somal tod ~ megelabiaste Rosert . sent
Other disorders of sulphur containing Amino acids... cei ont _oroooe I. Cystathioniouria:
‘Defect Custothonase Cyanide nitroprusside fest: Negative a cystourat Defect : Dibasic #8
transporter in intestine § renal tubules. 60ceb3eeassdampeseroenirrod’s tetrad : C -
Custiouria A - Akaptonuria. A- Albinism © - Pentosuria exertion in urine : C - Cystine (not
cysteine) © - ornithine Le Lsioe A frginine Cyanide nkroprussie test: Positive.
3. Cystiosis + Lysosomal storage disorders. Defect Cutie transporter ——> Custonosin
(product of Wugsosomal driver) Ts gene)
Blochamisty - 4.0 Mamow 6.0. 2022 Aitects : Liver > Hepatic faiure. Cenal > Renal faiure.
Cornea > Corneal opacity
Cute — Cupteine Specialised products from cysteine 010559 . Cupteine on decarbonation
gues setamercapto
ethanolamine.
a Co-enzyme & 3 Taurine : Conjugates bie acids. 4. citathione (esH.
S Cugtne 8 custome groups ined together by 3 sn grasps. Glutathione (GSH) 01.0856
• ksatrpeptide. @marroweditionBnotes 3 68s: Gamma glutamic acid + Cysteine +
Giycine.

• iis Gamma gutamy custeinyl gene.

• A pseudopeptide (sama. carboaylic acid forms he peptide bone.

• Rote part/business part/barking part & SH group of custeine. Functions of glutathione

• Amino acid transport: meister's cycle/gammaglutamy cule.

• Free radical scavenging,

• maintains 26C membrane integrity,

• Fioeps ron in ferrous state in hemoglobin. « ftioxdant.

• Conjugation In prase-1i xenobioti reaction.

• ets as co-enzume for vorious enzymes.

Biochemistry v4.0 Marow 60 2022

pr seo ii Fe
henson mines Glutathione ( co-enxyme ) Tbe sodosonae ~ggaaremres? Rares
acetoacetate Free Radical Scavenging (reduced) nop" HO « essa —~ Ss nnor eH (oxidised) t
Hime pathway, * Glutathione peroxidase A Selenocysteine containing enzye. + Glutathione
reductase: Flavin containing enzyme. MPR ASHRERRIAANSHO GOS
megs: ‘question : In which fem this clinical manifestation is seen ?
A Child is normal ot birth, sight developmental delay present. iter 3 yrs, he developed
severe * myopia, shivering oF iris is seen on | ‘examination 5
fans : Classic homocystinuria.
question : A 4 year old boy developed sudden onset of weakness of le side of body with
Facial devation to right Side. He is admitted in neurology ici with a suspected
cerebrovascular accident Hs let sided neurological weakness settied within a wee.
The treating doctor noticed a. waddling gait in the same boy, The mother gave a. past
history of defective vision due to dislocation of lens for which surgery was done. He has
poor
Blochamity «v4.0 - Marow 6.02022
scholastic performance in school for which he is
ping to special school. Further studies confirmed that there is decreased mineralization of
bone, creased length of long bones and sight curvature of spine to one side. what i the Tem
suspected here 7
A Cystioura.
© vomoeystinuria.
C. cystinosis.
©. Akaptonuria.
Question tA 4 year oid girl came 40 paediatric OPD with complaints of decreased vision and
poor schdlaste performance. 0/¢ ectopia lentis and multiple skeletal
deformities present. A test conducted © given in the picture. What s he diagnosis ?
Reser : Classic Homocystiuria.
Question : idenifu the biochemical defect from the description gen below,
• vomoptenemi.
pemetriioem.
• Wpocutenena.
a methene THEA Reductase defect. 6. methonine Synthase defect
€. Custathionine beta. sythase defect. ©. methyl Cobalamine deficiency. Queston ;
Lusosomal storage disorder uth CTNS gene defect
©
Biochemistry v4.0 Marow 60-2022 228 Proteins an Amino acids
36
A Cystiosis. ©. Custiouria.
c. cysteinemia.
©. Homoeystinuria.
Question : The amino acid with thioalcohol group. A methionine. ="
©. serine.
c. cysteine.
©. Threonine.
Question Vitamins needed in the metabolism of S containing amino acids are all except
[TY
©. Folic acd.
C. Ribotavn.
©. Cobalamin.
‘Question fi BRaER is SRraRation of of except A melatonin.
©. Serotonin. c. epinephrine.
Creatine.
Question : Al he folowing are functions of Glutathione except A maintain integrity of RBC
membrane.
©. Formation of methhemogobin.
C. Transport of amino acids.
©. Free radical scavenging.
Bochum 140+ Marow 60-2022 37
TRYPTOPHAN
Tryptophan : Chemistry 02:16
Indole Ring,
• romatic amino acid
• essential amino acid @marroweditionGnotes

• cath Ketegene # gheagene

“Tryptophan (rrp) ——> fraboie fate —

sreyptphan/ Tryptophan (FA 4 vayrleas /osigorans - (sarten/ wet hapfronjace
catabolic fate
hetogenie fate @uucogenic fate
‘Tryptophan : Metabolism - catabolic fate 00.08:27
Cotabole fate + /K/A Kynurenine anthranilate pathusayy
Blochamisty + V4.0 Marow 60-2022 230 Prownsana 37
ws -—
pine TEE orgs (ems contoming) | HP (Tet go Foe dic)
Form TFA Fupurenine | Fupmaiginajse * Require NADPH -0n Kpurenne
rupureninase + Coerayme = PLP (48) Raioe,
3-H fthwaniate 1
(Sucageni] Sate GfRTase Gate sting step NAD’ pathuuay Hetogenic fate GPRTase -
Guincinate Phospho Ribose TrancRerase * me, dem tere xanthurenc acd
4
exereted in urine
• Men — Petagra Ie syrploms.

Conversion factor © + omg of fuplophan CE 8 ac

Tryptophan : Metabolism - anabolic fate wa
• Tetrahydrobioptern *aeoPH i Tether Tes Ton
Blochamisy V4.0 Marow 60-2022 i
Degrades to 5-OH Indole acetic acid.
Functions of serotonin : + Neuro fransmitter. . Vasoconstriction, mood elevator,
temperature requiation gastro testnal ract motiy Site of synthess + Argenta®Sin cells. 4
Intestine, mast cells, platelets and brain.
• melatonin: Synthesized in pineal gand. Serotonin Goal AATARAR iC am’ methionine
son metry acetyl serotonin (relator) Function: + 6idgieal htm.
• Neurotrans@ifiarrowedition6notes
Carcinoid tumor/syndrome wm
Neuroendocrine tumor. Wi/ argentaffinoma. Tumor of argentadSin cells: i
1 Synthesis of serotonin | Niacin synthesis. Cliveal features : + Intermittent dasrhoea. +
Cutaneous Sushing due to 1 tachyinis. + Sweating | + Flctuating hupertension. + | Niacin —
Pela Ike synptoms. ; Diagrosis: 1 Serum serotonin,
1 Sve in a4 br urine (= < mg / daw. i
Blochamisy -V40- Marow 60-2022 232 Prteinsand 37 ‘Amino acids
Hartnup’s disease 0025:45
Defect in absorption of tryptophan § other neutral amino ‘acids from intestine § renal
tubules.
Intestine
defective ryptophon
fed > “Bue Diaper? Sacterial purine
‘eneyme
tedony compounds * indigobiue | + indican,
• Tryptophan in cells > | Serotonin and | Niacin.

me symptom iautaneeisiphatorensitivity Chotosensitive dermatitis due to niacin |).

Neurological manifestation (| serotonin) + wide based gait, intermittent atasa. Diagnosis by
Obermeyer Test: Test for indican, Treatment: * Supplement NAD" * Lipid soluble esters of
trytophan. Drummond syndrome + @onT-1: Transporter (oF tryptophard at intestine
coded by SLcuA. + Drummond syndrome : BOAT | is detective in intestine ue diaper
syndrome
o020:17,
Blochamiaty -V40-Martow 6.0 +2022
BRANCHED CHAIN AMINO ACIDS
Chemistry of branched chain amino acids 000150
eranched chain amino acids are Leucine : etogenic. Isoleucine : Hetogenic and gucogenic.
Valine : &lucogenic.
All are non-polar. Al are essential
Metabolism of branched chain amino acids ocos0a
Branched chain amino acid 1 Transamination | oe eranched chain keto acd cate branched
chan keto * eo mando Bara: ey con 3 FR dependent | Gee, oetugkogenaton Products Leucne
Valine
[Essen | 12 veto acid @retoiso xretod methy Coprocacsd valere acd valerie acid eranched
chain keto|acid dehydrogenase]
Biochemistry v4.0 + Marow 60-2022
234 Protas and amine 38 acids
Breeton metnacry Ta con
con con | hetogenic Proponufcon ety Con Giucogenic Ketogene
Branched chain keto acid dehydrogenase 00010
• mu enzyme complex (Simiar to puyuote detuydrogenase).

• Hos 3 enzymes ¢ head b ror rn ves secon —| a Diyrolpo trancacyose — €, 5

Dhughoipoamide dehydrogenase —

• coenaynes: Leon 2 Trae RRR Bote 5 Upoamde. «Fo s wor

MSUD (Maple Syrup Urine Disease) 01140

Defect in £0 Type 1A (MC) | Associated with eB: Tyels | thiamine pyrophosphate &, Type
Type nm Clinical correlation of msuD : * Neonates. + @ochemical defect : eranched chain
keto acid detydrogenase. Oecarbodjase enzyme (¢, enzyme component). Defect in oxidative
decor! ‘eranched chain keto acid 1 (accumulates) | x 80RD (defective)
Biochemistry v4.0 Manow 80 2022 feycon + Cincal features Feeding difficulty, Faire to
thrive. Lethargy, conwisions. Hypotonia wih bouts of hypertonia song. one 1 TEST oeging
rine (on refrigeration) maple syup/burnt
sugpr/cararmel (sme).
ages 1 eronched chan ameo acd — in 1 eranched chain keto acid | urine.
initro phen hydrazine test : Yellow edour precipitate. Rotherals test : Purple ring,
Mee er @marrowedition6notes Restrict branched chain amino acid.
Supplement thiamine.
Isovaleric aciduria 001810
13 week od neonate presents ith pure hypotonia, poor Suckling reflex. fn intern who
examined the baby collected. Some urine and keep it in refrigerator for one night. The next
dou a sweety smel was appreciated in the urine sample. she. ido ONPH test and got a
positive result
Ans : Male Syrup Urine Disease.
4.0 1 day old neonate presented uth vorvting, difficulty to Feed, lethargy. The characteristic
posture i seen. The.
Blochamisy v4.0 Marow 80-2022
francs amino
ili 236 Prieins and amino 38 Bis ae aaa
caramel odour is present. what is the diagnosis ? f. neonatal septeema. ©. msup, ©
Isovolers ociduria. 0 Orgone acidura. Hyperammenemia.
2. Assertion : Thiamin s used in the treatment of msUD. Reason: Thiamin is coenzyme for
the first step of catablcrn of cea,
A.A 4 Rare true, @ is the correct explanation for A
6.A§ are true but @ is not the correct explanation for A C.Ais true but @ is false.
©. Ris false 42 is true.
@marrowedition6notes
Biochemistry - v4.0 Marow 60-2002
ACIDIC AND BASIC AMINO ACIDS
Acidic amino acids Basic amino acids + Rspartic Acid — feparagie + vistidos + luton fd —
Gutamine igre + Lysine Basic amino acid wos vistdine frgnine Lysine essential | essental/
essential Semi essential Polar most polar Polar most basic imdozde | cuandnium | € amino
group Ghucogenie | Glucogene. Purely Ketogenic|
(@marrowedition6notes vistdine + metabolic function
Listing ———> Hetamine cod’
a Histidine | vistidose Urocanate i Imidazoione. propionate ’ : Formimion, akutamic. acid
(FIELD) THR Formimino folic acid eutamic acid
(= veto gutarate) — Ghucogenic fate
Bcchamisty -V40- Marow 60+ 2022
39
ii 238 Potensand 39
wn —m
© oefciency of THFA v 1 Formimion gtaric fed '
excreted in urine
• wstdne bad test
• WW FIG is excreted in urine —> Folate deficiency
3. Carmosine (f alanine + histine)
4. frserine (methy carnosine) S. vomocarnosine (6A@A + histidine)
Arginine and lysine 11:40
arginine + Metaboli functions. + Synthesis of
I. Agmatine — Antihypertensive.
a. Creatige rh Seng: bro & methionine.
3 urea ———> Arginine “3, orthine + urea. 4. Omitine.
5. tric oxide.
«wri oxide ‘endothelium Derived Relaxing Factor (E0RF) Free radical.
Gaseous molecule. Short half life : 0 J sec. frginine tric oxide, Nitric oxide + citrulline.
NoPH i Functions : vasoditor. i Penie erecton. Neurotransmitter Treatment of . Pulmonary
hypertension. 3 impotence (Sidena®)
Biochemistry V4.0 Marow 60 2022 39; 4 Remi Sidenafl = + (inhibits cemP
phosphodiesterose ) + 1 came
3. Angina pectoris igeery) nitrite — Nitric oxide.
• tri oxide synthase mono oaggenase S cofactors +L vieme a ew, (Tetvahudrobiopterid)
a naoPn amu Smo 3 sotorms + [ros = newrons 1) Nos — macrophages mark ARGH
riegRdent of caloum
@ Nos —> endothelal cells
Lysine * metabolic functions © Wistones are rin n arginine and lysine a. Putrefacton —
Cadaverine (polyamine) 5 Camitine Lysine + methionine
Acidic amino acids & amides 02128
Aspartic acid — 8) exira.- COOH in side chain
LL
utamine
Blochamatry + V4.0 Marow 60-2022 240 Proteins and 39 fm Sa
fparticasd Gtamcacd Asparagine Gutamine Non essential (08) Ne we we
lucogenic ucogenic @lucogenc Gucogenic Polar
Polar “uncharged polar — Aspartic acid Gutamic acd + Functions. + Function Pyridine 1
nacety aistamate a purine rey co A + Glutamic acid 2 Urea sypthesis i N- feety Glutamate a
sutathione (Gamma glutamy cysteiny eiycne) a ren Shtamate —— een
@marrowediionénotes ~~ © Glutamine: Functions LN3 of purine
a.N3 of puramidine: 3. Carrier of amino group from most organs including rain 4. Source of
ammonia (excretion of KK, —> Renal
regulation of Blood Pr) Enzume required : Gutaminase enzyme
‘Synthesis and catabolism of acidic amino acids 00255
Reporte fed (Transmination)
oul acetal iartate SUITES eto eutarate
tony
a chutamic fed Reductive amidation) @reto gutorate eon, ehutamate
Morow 802022
3 Aspartate Asparagine Asparagine
Sypthetase - coon -cowma utamine arp eutamate
altamate Glutamine alstamine
aptnetass dare catabolem scparagine fepartie fed ——— Oa acetate wl Ten cle Stamos >
Ships iho SRG iofeguarote | Canavan disease 0035:10
a month oid bow, born without compeations after an uncomplicated pregnancy, Saled to
achieve expected. developmental miletsones. As an infant, he developed oujtagmus and
poor muscular head control Physical examination findings were notable for generalized
potania § progressive mocrcephoby * eneyme deficient + Aspartoocyjase WN Reety
Reporte ——x—> Aspartic fed. reid * Clinical features Progressive maerocephaly.
Persistent head lag. Gevelopmental delay + on examination
Distorted mitochondria.
Biochemistry + V4.0 Marrow 80+ 2022
39 Acc
we
acs 242 Proteins and Amina acids
39
Severe leukodystrophy.
1 Kacety aspartic acd i blood, CSF, urine. Questions © 1. Fatty acids cannot be transported
fo mtochondiria, sine is deficient. why ?
ns Because carting, hich is the transporter of fatty acids, & derived from lysine.
a. stones are posituely charged. why? 0s +8 they are rich in basic amino aids ike avgirine
§ aioe.
3. Sidenal s used in he treatment of impotence? fins. Inhibits cyclic GMP
phosphodiesterase and thereby increases cyclic amP (and messenger of nic oxide
(derived. from argeine).
PEL RR AR mn debian wn? fe Fle tocar 6 omar rou 50 when
there is Solc acid deficiency, FIGLE gets excreted in urine.
meas. 1. The amino acids that form oxaloacetate are A. Asparagine { Glutamate.
© fisparagine { Aspartate.
C. aiutamate § amine.
0. Glutamine { fsportote.
an infant normal at birth later develops progressive mac- rocephaly, severe hypotonia, and
persistent head lag, me! scanning of head revealed striking Vacuization and astro- cyte
swelling in white matter. acetyl asparti acid present in the Hood § CSF. what & the
diagnosis ?
A Alexander's disease.
Reyes syndrome.
¢ Canavon disease.
©. Toy sachs disease.
Biochemistry V0 Marow 60-2022 39 Acidic ang Basic A © Sandnoft's disease acids
The rtragen donor For asparagee synthetase & A mero on
8 sutamee.
C fspartate.
© automate.
Tie donee 0 Som goup in ne carbon metabokm a sere.
o Tuptopran.
C wets
0 args.
5 Urocanate an termedate in whch amino acd metabolism? R proine.
© wate.
Threonine.
© frgne. @marroweditionénotes
Ornithine derived from uch amino ac? A proine.
8 vstdine.
e Trveonne.
0 frgrine.
Blocnamstny +
0: Marow 60-2022 244 pres andameo 40
MISCELLANEOUS AMINO ACIDS
Entry of amino acid to TCA cycle/ Anaplerotic reaction . ooorer

5 piyuate to oxaloacetate
• Wydroxuproine.

• Serne.

• cutee.

• Threonine.

As alarine to pyruvate to oxaloacetate :

• Tuplopran
Oirecty to oxakacetate :
• Asparagine — Aspartate —> Oxalacetate. as alutamale o Ketogutarate.

• Hatin.

• prolie.
• si@tmineeditionbnotes
• frgeine.

fs sueciny CoA:
• isoleucine.
01031
Sarcosine — N methy glycine. etaine — Trimethyl gycine. (2x of homocystiouria Choline —
Trimethy etrandamine. Ethanolamine —> Serine on decarboxjtion. exgothionine —>
Derivative of histdne. mercaptosthandamine — Cystene on decarboajotion. ©06A >
automate on decarboxytion.
Biochemistry -VA0: arow 80-2022 + camosne — f any vidoe. + Prserine — Carnosine on
methjation + Vemocarmesine > 6A + vctdine.
Urine odour in various inborn erros of metabolism 001350
Inborn errors of metabalem.
tar aceon Cuge 1 Voukirseura Isovaleric acidemia. - 5 Sgro meng caturee magle syrup urine
dsease [ei Spina nperchincama [sowie mutple carbosgase defency | Tom cot ure asta ure
deease cote cabbage rope Ine [— @ Mal GOES Tremethjameur (Fen odour) | Gotten fen
Tyosnema. ooted cabbage ota + enzyme defect : Trimethyjamine oxidase. (Flavin
dependent monooxygenase).
• Trimetnjamine is not metabolised. > Smell of rotten fish.
• Bx Restrict dietary intake of trimethyjamine (choline) containing foods. (eqas, nuts,
green leafy
Q. The amino acids that form Oxaloacetate ore : A Asparagine and Glutamate. ©. Asparagine
and Aspartate. C. Glutamate and Glutamine. ©. Glutamine and Aspartate.
Biochemistry +V40- Marow 80+ 2022
a0 i 2
206 Bomnereics 41
KREB’S CYCLE Concept of TCA Cycle B 00122
16 8//A TCA eye (Iriearboaie acidb/ciric acid cyde/ vrebs cycle.
Carbohydrate, protens, lipids. N | feety) CA CAC] Completely oxidized enters TCA cule
releases : a C atom 05 CO, NADH, FAH, — enters £TC to produce ATP.
Site: organs with mitochondria. Al enzumes of the TCA ce are present n the mitochondrial
matric except succinate dehydrogenase. (@marrowedition6notes Overview of The TCA
Cycle ao0s24 Regeneration of Oxidative oaloacetate decarbojation Cotolgiero | FER
Oroloacetote 2)

I I I I Citrate (oO) 1 | |
“oo
30+ Mamow 50+ 2021 a x 4
00807
Ovaacetate + feet Co crate space Dom ID Cte,
Ee nn BN ad
“Conan ror mate “Sep proces
Oebugraton, Seywaten woe pre
WN Chote feontace
se on vemos Sse
(Goer ov (mepias
mis glutamote—> enon
Succiny! Con —> Heme oxaloacetate —> elucose
Aropleurotic reactions : Wes a Siling-up reaction Reactions that replenish depleted
intermediates of the TS Pypanie certs
‘most important reaction : Pyruvate ——> oxaloacetate.
1_
TCA cycle & cancer - Oncometabolism 0033.95
oH Normal: socitrate —> ati —> succinyl Con
IDM mutant : ae —> a hydroxyglutarate (oncometalbolite)
J
Inhibits Ter a gene (epigenetic modification
Increased OM" methylation § histone modification
changes the epigenome
Biochemistry «v3.0 « Manow $.0 +2021 “ite
Cancers related to mutant a hydrosyghutarate ¢ + Chlangjocareinoma, gfoma. + eute
myolod leukemia, Yar iaus sarcomas.
rugs inhibiting mutant 10M: Sidenbs Cevasidenibs, Wosidenibs).
Mutation in SOH complex 8 1 0) causes : Fomilal gjoblastoma. Famiial pheochromocytoma.
Vitamins. in TCA cycle
• Con: Pantothenic acd
more,
© nore,
«+ areon:e,
Regulation of TCA cycle Eegplatony step |
| Chmtespiuse @marroweditionénotes a tocitrate Dehydrogenase.
3 ane Dehydrogenase.
4. Pyruvate Dervdrogenase. (especially in brain).
1aTP/60P § 1 NROHARD ratio + hits TCA cue tegiaiony enzymes.
Blochamity -VA0- Marow 60 2022 G2 -— CHEMISTRY OF NUCLEOTIDES
Chemistry of nucleic acid 00125
Two types of nucleic acid DNA and RAR.
The nucleic acids are made up of nucleotides. The nucleotide s made up of
nitrogenous bose (purines § purimicines)| pentose sugar (ribose, deoaybose] Phosphate
group.
itrogenaus base + pentose sugar=
Neudleoside.
Purinebases 000330
a heterocyolie ring structures with rrogen positions ok 13,19. Purines : Adenine and
guanine.
minor pufines XaRNe, RjpoRaNEne, and uric acid. u n Pure » © amino purine a amino 6
oxopurine wn o Me § NZ Hen i S > A t u i N ' i i adenine @ Guanine ©
Boch +A -Marow 80+ 2022 43 Nigger pyrimidines
Single heterocysie ring structure with nitrogen positions at 13. Pyrimidines Uracil CANF
only), thymine CONR only}, Cytosine.
_
af dioxo a, 4 dioxo S 4.010 4 amino
Pyrimidine methyl pyrimidine a “o> ~
I4
ured GD PP x only
Ra ont ‘both DNA ana RNA
Nucleoside formation 00:10:29
Pure: BW glysosiie bond between ¢ of pentose ‘uagar 4 9 o pure. C10 pentose suger 411 of
pyridine. Pentose suger. On group ic present ata 3 positon ribose. Ron group is present ony
in postion deoxy ! a
Blochamisty “V4.0 - Marrow 60-2008 252 Molecular Bicogy
43
Nucleotide formation 1812
Addition of PO group by ester bond to the S! C atom: Nucleoside monophosphate.
Further POA groups are attached by acd anhydride bond. an acid anhydride bond is broken,
energy is released.
Dinucleotide formation 01637
The two rucleotdes are joined by a.3'o <' phosphodiester
bond. Betueen the 3 On group fo the § PO4 group. (@marrowedition6notes
The nucleotide with a. free functional group ok < positon
First nucleotide Chead endl.
The nucleotide with the free functional group at 3’ positon : Last nucleotide Cail endl.
These functional groups are ionizable — exhibit Polarity. 4 convertion me eas SEsHhce of
nucleic acid is written from So 2.
Nitrogeneous base
PO—CH, ~ pentose
Biochemistry -V40- Marow 60-2022

(@marrowedition6notes
Blochamatry + V4.0 Marow 60-2022
© Em 254 Bioenergetics 42 EE ELECTRON TRANSPORT CHAIN
Basics of ETC 00128
Oxidation Loss of electrons. Reduction Gain of electrons. Redox couple Compounds that can
ex in oxidised as well as. reduced state 23: N/R, FEO FROM, Fron/Fra, Redox potential
Ability 4o transfer electron/gain electron. areater the redox potental greater the abity o
gain electrons. Electron moving from luser to higher redox potential exothermic reaction
Components of ETC 0040
STL order of redox potentials.
Complex 1 NROH-Coq oxidoreductase + Ouse NAOH, redhices coenzyme @. + Components:
Fw § Fes. Coenzyme o/ubiquinone/a,, + mobie electron carrier Complex: Succinate Q
oxidoreductase. * Oxdises Succinate, reduces coenzyme Q. Fes complex. Complex 1: G-Cyt ¢
Oxidoreductase } + Components : Cyt b, Cut Giese Fe-s complex. Fo compen icutc asec - +
Wreversivle complex + Components : eme a, a, / Cyt a, CuR-Cu centre
Bochamaty -VA0- Marow 60-2022 ‘Complex V oosz.t2
Alco Known os ATP synthase complex. F, subunit: * Spans the inner mitochondrial
membrane. + made of 10 € dise proteins. * Hydrophobic * proton channel F subunit made
of 9 subunits : 3 a 3B ATP supthesieing unit Y (rovie) ,d and é
@marrowedition6notes
‘Oxidative phosphorylation 002117
‘Outer mitochon merrane

=;
or ray L) a} es woe arp OP
Biochemistry “V0 Marow 6.0 2022
42. Elec “War
— 256 Bioenergetics 42
Oxdation: * movement of electrons, leads to release of Sree energy. * Pump to
intermembrane space. Phosphor ation A proton motive force is created (dfSerence i
potential due to pumping of protons). 2. W moves to mitochondrial matrix cus, subunit 3.
Rotation of Y subunit. 4. Conformational change in § subunit (nding change mechanism by
Paul Bower) * K0P > ATP. Oxidation is coupled with Phosphoryjotion by a proton gradient:
ROH pumps 10 FROM, pumps 6 W ROH generates a5 ATP (0 ratio = 29) FAH, generates iS
ATP (PO rato = 15) This is explained by the chemiosmotic theory by Peter mitchell.
Inhibitors of respiratory chain ooze nhibitors of electron transfer oxlative phosphorytion
eisitors of egctron transfer
) rn Ro "X purovertin Br aop-ae 2 Y
Gitano Chnioicy pop TP ? Lewsie)/
eboitors of
Dmercapral “Bokmyne A oudate prosphonyatin
Chemical uncouplers : + Dinitrophenol NF + Dintrocresol (NE) + Fluoro carbon cyanide
phenyhydrazone (FCCP) + epic in high dose:
—
Blochamisty -VA0- Marow 8.0. 2022
— © SRE a Biplifdmasgies: ‘Thermogenin/uncoupiing protein-1 (UCP-D : non Bite passes
Found in brown adipose tissue (chubby cheeks in neonates). There is oxidation in the
tissue but no phosphoryjation (action of thermogenin). movement of ie wi vt Fae Ws Ce
behind prevention of hypothermia. in newborns. emis Se REE uated Chub:
High energy phosphates 003759
Produce free energy > 7HCal Examples : Phosphoend pyruvate. Carbomou phosphate. eee
Creatine ProsgitarroweditionBnotes ATP oP + Pr ATP Ame + 01
Biochemistry V4.0 Marow 60-2022 258 Molecular Biology
44 se asta METABOLISM OF NUCLEOTIDES
Purine metabolism 00.0057
De Novo Purine synthesis + Occurs in most of the tissues (cytoplasm) but mainly in the
liver. De Novo synthesis doesnt take place in # Brain, erythrocyte. Leukocyte. one marrow
They purely depend on the Salvage pathway, Contributors to Purine ring : The foie atid
component in the
purine ring plays on important role in cancer treatment and. its deficiency can cause
defective DNA synthesis
oe @marrowegdfionGnotes
i1J
Blochemisiy “V4.0 Marrow 8.0 2022 ie fo
Roto gong fopartate L | Glutamine
• ppp supihetase : reimnary step Regulatory step
• PRPP can go into different pathways | SaNoge pathy NRO" sess. Pyrimidine synthesis.
• PRP Glutamy Aminotransherase : Committed step of puree sthess Recpintory step Rate
mite sep
Salvage pathway 01113
Purine nucleoside and Purine bases are recycled to Purine
ruseotdes @marrowedition6notes
cans, 1

Saves energy a. efSective recycling. 3. important in organs without De Novo Purine

syntheses. Donor Sor Ribose and PO, : FRPP, The reaction is called
Prosphoribosyjation.
Blochamisty -V40- Marow 80 2022 =. 260 Nokes 44 EET
Biology Donor Sor PO, * TP The reaction s called Phosghorjation.
Reactions . bosyaton fu pup vepeTase iyponanthine Guanine PETase, a vupocanthine/ anne
tose monephosphate! uancine manaphosphae. seerase
bo. Adenine —— Adenosine monophosphate. fer pe
HEPETOS, pposphorioosy transferase
Site: Dietary Purine aegracea Tv restr. endogenous are degraded n Live: ‘end product : ric
acid. Organelie : Cyfoplasm. Steps of purine catabolem Adenosine Adenosine X SCID l=
immunodeficiency dsease ose x wanosine N Purine nucleoside ow” ribosy transferase ei ©
wypoantrioe wanine xanthine avanine
og
X oddose | WO, uric acid
Biochem VA Marow 80-2022 e— pale fo + 5010 (Gevere combined immunodeficiency
defect) dia sabes dl oi tough a wbcind: * Immunodeficiency disease © ta ices oy afore il T-
cells are affected § B-cells are normal. Sei Sain ode debe aio pha oS A PRES
Xanthine oxidase reqpires molybdenum. Allopurinal inhibits Xanthine oxidase.
Disorders associated with Purine metabolism _ 0025.46
HePRTase deficiency I. Lesch Nyhan syndrome
x Inked recessive disorder: iochemical defect : HePRTase completely deficient.
vpoxanthioe/~) _y | we/ i cuanie ) ame i Purines accumulate — Increased uric acid
(Hyperuricemia) Clrical features : + compulsive self-mutiation * out
Blochamisy V4.0 Marow 60-2022 262 Molecular 44 “ee ara
• Dystonic movements.
• megaloblastie anemia.
• Neurological deSet
Treatment: * fAlopurinol (inhibits xanthine oxidase) + High Suid diet. + Aalnize urine.
a. Kelley-Seegmiller syndrome HePRTase is partially deficient. > 1S-af activity +
nerrase deficiency
seerase JP satenine —X-> eene oxdaton| | pRPP PP

a8 dihydroxy adenine 8,8 dihydroxy adenine causes : Severe erystalluria. causing

Ure eciderystaks st metatarsophalangeal pint Chronic gout
So)
TRAC ee etna Ci anpenn” Mea
subcutaneous tissue Cong As
Biochemistry Vi Marow 80+ 2022 “* Nec es Biochemical defect: + In primary gout Enzyme
defect + ‘activity oF PRPP synthesis > Purine» ture acid ‘ retivty of PRPP Glutamy! _) amido
transferase Causes : Lesch Nyhan syndrome. Glucose-t-phosphatase deficiency : von
eierke's
disease (Type 1 <0) + mcezoery ot oY teen turnover cecreased excretion of ure acid Causes:
J + Lactic acidosis marerey + Renal faaure Fagravotng Factors of gpUt! 6 ouretes (Thazided
fcohoksm.
• Inereased Purine intake (pease dition notes
• Junk food : Fructose rich > Hyperuricemia
Symptoms oF cout : o—l— m3 feute Chronic + peute infammosory a" orthritis + Unie acid
nephroithiasis
Diagnosis of Gout : Aspiration § examination of synovial Suid ‘most commonly affects the
first metatarsophalangeal joint bbecause the acid erystalizes commonly at the cold,
peripheries. Treatment : + Allopurinol i + High Suid diet. i + Aalinization of urine. +
Anticinflammatory drugs. * uncosurie drugs Probenecid). * Prevent aggravating factors.
‘Blochamsty + V4.0 Marow 60-2022 264 Molecular Bicogy
SCID & Severe T-cell defect
SCID + Severe combined immunodeficiency Defective enzyme : Adenosine deaminase.
most common cause : Defect n gamma chain of a, Non-homologous end joing (HED
(defective or repar) & also a cause.
oth B-cells and T-cells are affected.
The first disorder +o be treated bi gene therapy (by or. French Anderson : Father of gene
therapy.
enyme Replacement Therapy (£27): Polyethylene gues! modfed AA is aso a treatment
modality For SCID.
Purine nucleoside phosphoryjase defect : Severe T-cell defect and 6-cells are apparently
normal
Pyrimidine metabolism os
Ste : Lv marrowedition6notes organelle : Cutoplasm ond mitochondria. 0a + slutamine etd |
ren Casbamoy fo, (cho enzyme nt rangi 1.7 (Gna plpeptde) Aspartate ——-—
Transcarbamoase. Gre) carbamay i J Ohidroorotase: J ‘hydroorotic acid NADY |
Diidroorotate Jes
oro asd a oll (Seno fobpept)
(0+ Marow 80-2022
He Po [ome decarsoapuce uridine monophosphate (Ue) : Pyrimidine nucleotide
ATP | Dribonuceotide bd Jae
os duop
ea mye Sor, FL
S-Fluorouraci inhibits Thymidujate synthase.
methotrexate inhibits Dinydrofolate reductase.
Orotic aciduria o0s404
Defect in the De Novo sythetic pathuay of purimidines.
tol roi,
Terre stro RIES Rte aprase:
SAAR Ss Ko 0 BEER
ceang densa ying + dered ab
ph Rt
Type 1 Orotic aciduria.: Defect in OMP decarboxyjase only.
end products of Pyrimidine catabolism cytosine rac Thyme
N Ls
B-ranine B — Amino sobutyate | water soluble Therefore they do not crystallize and no
symptoms seen. Conditions causing Orotie aciduria : * Type I huperammonemia :
ore — 1 carbamoy PO, (mitochondria)
CP (cyfoplasm) — Pyrimiine synthesis
orotic acid
Biochemistry + V4.0 Marow 60-2022 266 Molocuiar 44 Bion [oi res + Reye's syndrome :
Defect in mitochondrial enzymes (Urea. cycle enzymes). Increased CP leads 4o orote
aciduria. Pseudouridine : 115 Sound in the pseudouridine arm of 12a. Uridine = uracil +
Ribose sugar “ie rpetourdine = C Puy + C) Pentose sugar.
Pyrimidine Sahage pathway ¢
Rye ne 5
x | Pyridine nucieosite
Pyrimidine base
oo 1 merit hd presente pith Selore o rive, macrocuic upochromic emia. Bone mari
éxamieation shied mecitiost No respense fo iain 83 o Foi fed of ron what is your diagnosis?
rs. rote Acid,
Blochamisty -VA0- Marow 80-2022 STRUCTURE AND ORGANISATION OF DNA
Introduction oo Dark lacy of DNR osalind Frankie.
Structure of uA is called as Watson § Crick model of AR discovered in 1952, For which
Wadson § Crick wos awarded Nobel prize in the year 96a.
salient features of structure of on:
Two Polydeoay a other in right handed drecton.
Hand rods Formed by sug» phosphate Steps Formed by bases.
ft-parallel arrangement of 2 strands of DNR + One strand 's aluays in $ > 3 direction. whie
the other strand in 3 > direction.
Diameter of DNA is anm.
Base paring rule
Adenine ( abuaus pairs with thymine (1) by two hydrogen
bonds. Guanine ( pairs with cytosine (¢) by twee hydrogen bonds.
Rederine always pars end roe Brame
Ferd ini
Cs oo ree =n
re ann vou
Bochamisty -VA0- Marow 602022
a5 268 Molecular 45
soo ET
Chargats's rue: It states that number of purines = number of pyrimidines. Chara rule
Purines mmm pyrimidines ase stacking OVA + The bases are aromatic and nonpolar they
stack on ‘each other by vanderwaal’s forces called base stacking, + The vanderuuaals
hiekness & 3487 average of 105 eps hm us bp tees © Dameter : a om.
minor § magor grooves They are the sites for OXR Pom; eels SR TES
Different types of DNA 00927
o Forms of onR * #,6,,0, > Right-handed. * 2 Left-handed.
• The form of DNR preferred in a solution devoid of water is 8 form.

• The back bone of DNR § 29 209 in 2 DNA.

• The onR found in low sal concentration and igh

degree of hydration s & form. + The ox found in high salt concentration s A form.
Blochamisy V4.0 Marow 60-2022 ona + 2 DUA & Sound predominantly in certain regions
where. purimidie alternates with purine (most commoniy C 1 o * DNR complimentary to
mRNA: CON. + Chimeric DNA is formed when DVR of interest jos with Veckoc oa + + 2 form
found ot the ends of the chromosome. Mitochondrial DVR © + of celular DNR. +
mitochondrial DAR is double stranded, creular and has about 6569 bps. + Muman
mitochondria has 3-10 copies of DR. + mitochondrial DAR encodes 37 structural genes for
RNAS (SRNR and 13 rene). a mitochondrial teks. 13 proteins of €TC (electron transport
chair). © x10" bp present in human diploid chromosome. 13 proteins of ETC are: +
Subunits of Compigg Inarroweditionénotes + Cyl of complex il. + 3 Subunits of Complex V. +
a Subunits of ATP systhase. ut of 57 subunits in ETC 13 are by mitochondrial DNA thot
constitutes 19%, Unique features of mitochondrial DNA © + Mitochondria, has a unique
genetic code; Only 43 RAS are involved in translation in mitochondria.
codons | ulear code | mitochondrial code
wn oleucee metnonine
wn stop codon Trprogpan
eh ee egos Stop coun
• mutation rate s very high because i L No mtrons. i 2 No protecte histones.
5 No effective repar enzymes. 4. Hs exposedt to omuggen free radicals, generated by
oxedative phosphorylation.
Bochamaty -VA0- Marow 60-2022 270 Molecular Biobgy
• mitochondrial DAR has non-mendelian tupe of iherttance (cytoplasmic inheritance)
known as matriinear nheritance.
Denaturation of DNA 001908
Ao Known as meting of Due
“The process by hich tuo strands are separated in to
component strands.
Features of denaturation :
• ereaking of hydrogen bonds.

• Bose stacking also disrupted.

• phosphodiester bond is not broken.

• No Covalent bonds are broken,

• Primary Structure s not altered.

• Secondary and tertiary structure altered.

• Viscosity is decreased,

Ee So cated hperchvomieity DRA absorbs dom uv light Denaturation dentifed based on the i
the 240% of Vlg passed rough the OR absorbed or not
meiting temperature (Tm) : Midpoint of range of temperature over which the fu strands of
DNA separates.
Features of Tm * ase composition. * Salt concentration. * A10-%old increase in monovalent
metal ion concentration increases the Tm by ls degree Celsius. + Formamide * Destalbiizes
the hydrogen bond.
Positive supercols : NR 1 wound in the same direction os of NR hel ie, clockuuise or right
hand.
Negative recoil + DNA is wound in he opposite direction as hat OF DAR helix Le,
anticlockuue.
Biochemistry VA Marrow 80+ 2022 pr Ser Se
Toporsomerases : + 60 Known as nicking releasing enzymes. * Relieves topological
constraints of DAA. + Types: Tye Tea mares sje stranded | mates nes i both the nen nthe tas.
strands of tre oun TP not needed |g needed. Organization of the DNA 002730 Prokaryptic
DNR is not organized. Eukaryotic OVA a highly organized structure. Levels i. Du double
heloc it. 1onm chromatin bri. ii. 200m chromatin Rbril.
4. Nuclear scala Unfit SEMRSABRb tes +. metaghase chromosome.
lonm chromatin Sri + + nade up or Nucteosomes (ucleoproten compos. Nucleosome
consists of a histone octamer and DNR wound around it. + 175 turns (4a tums) in a histone
octamer. * Left-handed direction. + Solenoid structure. 300m chromatin Rbril : Has total of
© nucleosomes.
vistones : + most abundant chromatin protein. + vistones are positively charged (DNA is
negatively 1 charged). lore bound found between he ONA and the i protein.
• imino terminal of #he histones i rih in basic proteins Ike arginine and sine, hence the
posite charge.
Biochem -VA0 Marow 80-2022 272 Molecular Bioioay
tno
as
• Wstones classification :
[oe rrr
Present i the histone | make rc in both the sirande. octamer.
HoH ore the coithubones. [ven
The Inker OKRS connecting the rucleosomes has a beads on
string appearance? Around 35bps are present in the leker DNA.
Euchromatin 0380s euchromatn vetererramate | Less densely packed. Densely packed. |
Transerptonaly actve. | Transerptonally nactve. |
Chromatin stains fess, densely | Chromat stan densely.
perm@Eavdate [1071 61adg8ce drvomatin.
Types of heterochromatin + Constitutive : Centromere, tekmere. + Facultative One of the x-
chromosomes’ in females.
Q.1F a sample of DAR has adenine a¥% what wil be the amount of guaninge present ?
aa
bash.
cach
dam
es
Q.uwhich is more susceptible to alah denaturation, NA or DNR 7
18 is more susceptible to alkali hydrolyse han ORR. The Sree hudrony group is responsible
Sor rapid aka hydrolysis.
Biochemistry + V4.0+ Marow 60-2022 DNA REPLICATION
Introduction 000051
oetkon: OAR replication i the process by which copying of base secuence present in the
parent strand to the dougrter strand Hares place. helps passing the genetic formation 0 the
progeny. Salient features © tases place during the ‘<' phase of the cell cule. + Both the
strands of DNR act as template. + Wolf of parent strand is conserved (semconsersatie
modeD. Base pairing rue i alias obeyed. New strand always synthesized in §->3 direction.
nA replication i a bidrectional process. 1s a semi-dscontngays fTRseRRIORTHG oF a leading
strand (continuous 1 a lagging strand. continuous. pp Sheps of oa replication” 1
Wdentification of origin of replication (Ord. 2 Urswincing of DNA 3. Formation of
replication Sork, 4. OnR sypthesis eading § lagging strand).
Identification of Ori, unwinding § formation of replication fork :
renal aon org of repieaton | TOO Newoun mS Jen et So Geptcaton iste 7 eepicaton fom
Biochemistry V4.0 Marow 60-2022 274 Moscur a6
Boos TT
IdentiReation of origin:
Ort 6 dentified by certain proteins
Ori 60 + Ori binding protein. This protein binds 4o the origin of replication
Near the origin of replication, an AT rich sequence & present. The AT rich region n
eukaryotes are known as CUE (nA unuinding element.
The Or binds to the Ori and the AT rich region unwinds. The 556 (singe strand binding
proteins) prevent the reliking of reannealing of separated DNA. In humans the 5565 are
Known as PA (repetition protein ©.
Once the DNR is unwound, a bubble likes structure is formed. with & replication Forks ok
each ends.
DNA synthesi:
Leading strand template : The strand runing in the 3 fo 5’ direntin~
Leading strand supthesis |
• "step RNA primer synthesized in 2) drection; mediated by enzyme primase.

• a step Synthesis of new DR Geading strand) n

continuous manner; mediated by ORAP-1 (in

Bochamisty -VA0- Marow 80-2022 © ie provaryotes) New strand from §' to 3' direction.
Laggng strand synthesis : +1" step RNA primer synthesized (by enzyme primase). + a step
Okazaki fragment (short strand) synthesis; mediated by DRRP-il. + 3" step + Removal of
primer 1 ing of gaps; mediated. oy OXRP-. + 4" step Sealing of ricks by DNA ligase (reauires.
ATE). reprint
ron copacemant os SET
eB O) OOS
so apiece. Sri
Enzymes of DNA replication & their functi
02240
• Topoisomerase : Preg@nistorsonalstraio los
• Welcase * Unwinding enzyme along the direction of movement of replication Fork.
ATP driven enue.

• OVA primase : RNA primer synthesis

• OnR lgase : Seals the nicks.

• OVA polymerase : Rots only in 5-3 direction § requires primer.

oR polymerase : Acts only in $3 drecton. Requires primer.

Prokaryotic J euranyote:
‘OA polymerase | ONRP-D. oneea.
OMA polymerase If ONRP-ID. | DNAPR.
on polymerase 1 (onAe-1D. | ONRPY.
onset: + iso known as Kornberg's encyme. + Discovered by Arthur vornberg in Ecol
bacteria.
Biochemistry + V4.0 Marow 602022 276 Moecuar Bicogy
• Wenow polymerase/Sragment : ONRP- from which 5-3 exonuclease acti is removed.

• Actions Removal of primers, DAR proof reading, RA Tepan runction (aon) ONRP-1 =>
DNA proof reading § OR repair Grinor). ONRP-1I > Leading strand synthesis, Okazaki
Sragment Suythesis, OA prook reding, most processive NA polymerase.

DNRPE > Fs the major DAA repair enzyme.

ONAPY > Required Sor mitochondrial synthesis.
ONBPD > Required for lagging strond synthesis.
ONAPa > Responsible For primase activity,
on8Pe > Required for leading strand supthes.
ONPE, DRRPY, ORRPD > Have proot reading mechanism.
DNA - damaging agents, defects produced,
repair mechanisms & associated disorders 03311
Tor | odes | eapar — Fubar | __ arg [Ewe [un es csreted wh se adaten Parogenows | canbrad
rune decency | ow [seme |edpony | G00. we [comiee 15 tava reangactasa Gcvr/ovea ender.
men trea scare. ow. Bons pcre | | werner sperm. | ota Tromeon
esocated dicts
£
iseoae | weroderma paren. cucnaupe spore. Treromagiogng,
Wigner
ppg
Veredtorycan-pogone coon cancer arte a Lape
Biochemistry -VA0- Marow 60-2022 46 oes
• There is loss of genetic material,
• 18s the major mechanism in humans.

• There is exchange of genetic materials, nally leading 40 correction of the defect, *+ 8s

the major mechanism in yeast. xeroderma pigmentosa : Helicase enzyme is the
defective enzyme.

Telomere & telomerase

Telomere + + 18s. me ens or me chromosome’.
• Has ‘TREE repeats)
end replication error: * Once the primers are removed, i leaves a. gap at the s! lend of the
daughter strand. + 3) end of 0 parent sfURHis GUKTEPIGREAO IOS *+ Wot corrected, #t
may lead to shortening of OAR (generation after generation).
Tainan sete! + Also Known as terminal telomere transferase.
• 185.0 ribonucleoprotein,

• Ws.0 protective enzyme.

• ik corrects the end replication error in certain cells.

• tehas an intrinsic RAR primer, which is complementary to the sequence that is present
at the ends of chromosomes (TTAEE repeat). This helps it to easiy catch on 4o the ends
of the chromosome and corrects, the end replication error.

• Ithas reverse transcriptase activity,

• Absent in somatic celts.

• Present in germ like cell, ster cell, cancer cell,

• Increased telomerase activity causes cancer.

• Decreased telomerase activity couses premature aging

‘Blochemistry “V0 Marow 60-2022 278 Molecuar a6 bee -_ (progere). * acts as target for
drug development for anti-cancer drugs. © ayflex mit > Abter 750 cells division replication
potential becomes nl The is because of the absence of telomerase actity.
Difference between prokaryotic & eukaryotic
DNA replication oose21 - Seong on Er oo fremase ONA synthesis onAPe eading | strand);
upd Gaping strand) Removal of | onset [ease n/ Sap primed MAITOWEIONSNOIES |
gnoruciease
Boch, V4 Marow 80-2022 TRANSCRIPTION
‘The process by ihich en is synthesized fromm OAR.
Salient Features oo01:39
1. Only | strand acts as a template.
enn Py tempat strona / minus © sand / fericence stand
hone
Shaka coding Seond/ Pus srond/ Sente rand”
2. eA & synthesized in the s) 2) direction @ecause RAR polymerase can act in the same
direction ony.
2 No pemer ie required
4, The newly synthesid@areancemudaiensttetsame as the coding strand (exception Tis
replaced by W complementary to the template strand.
RNA Polymerase (RNAP) 00:08:51
• Prokoryotic RNAP + only | ype RNAP muiti-subunit eneyne.
Sa sre subunt)
Srp acme tnagre @
Sigma. subunit: Helps in the binding of RNAP to Promoter site.
B - subunit: Catalytic subunit
2 types RARPs. Classified based on its sensitivity ‘ouards a. poison in mushroom Comore).
Wobeenzymne
Blochamisty -V4.0-Marow 60-2022
47
280 Molecular a7 Boy [eer awe | aomantn | Prods sensi — RNAP [Least RNA (most
abundant) anor [wget ers, mane, ; coon cans een [itermedate | 1@a8, ssrean, " Jeertansnaa
Promoters of Transcription 00:12:38
Promoter : Conserved sequence in the coding strand that specifies the stort site of
transcription (boxes or elements). s pomoter y FELL ITT >. bedghand Orr promotere—|
LLLP PTT 0 10 fogn y Canearsees upstream + ~ Downstream @rtroweditionnotesiements
Template strand
‘Bacterial promoters : * Prionow bg: = 10k * Te box: - 35bp Pribeous box s 3 sep -obp)
• c——
eukaryotic promoters + TTR bow/ogness Box - asbp + caT box: Tsbp. + ecko CanThox TER
80x wosaESS. 80
sso
Tew mw ——
Biochemistry V4.0: Marow 80-2022 a7 ih %
Promoter less, sequence : No promoters. Additonal promoter elements :
• tor (vtiator sequence)

• ope (ounuard promoter element) Promoters are upstream elements, and they are not

transeribed. ‘exception : Intragenie/Dounsiream promoter — Transeribed. Example | NAP

1.
Transcription Cycle 02040
I. Template binding § closed promoter complex. a. Open promoter complex.
3 Chain nation § chain elongation.
4. Chain termination.
nL WOOL = EO, x @mamoweditjonénoles
rnp mor XO
BRIA
Termination: + p dependent + p independent p dependent termination: +p factor binds 40
the &xR — DNR complex. +p factor & an ATP-dependent RAR ~ DNA helkase — detaches
the 2a from the ORR. i p independent terminaton : The templote strand has i termination
signals (6¢ ih region and U rich regiord. C rich region forms intrastrand base pairing, thot ,
hairpin structure. This is followed by the U-rich regon that forms bonds. wth #he A region,
which & comparatively weaker.
Biochemistry V4.0 Marow 60+ 2022 282 Molecular 47 Biology
Hence, Ru detaches from the OMA. Post - transcriptional modification (PTM) oozrts
Proaryotes : All ENA undergoes PTm except meNR. eukaryotes : All AN undergoes PTM.
PTm in mann 7 = methylguanosine capping : ft the 5! end —> 7 ~ methy) guanosine cap «
added. Steps
• GTP ic added to the 5! end by euanyly! transferase, in the nucteus to the mans (o/w/a.
Primary tronseript/ nena theteronuclear 2NAD.

• Inthe cytoplasm, methyjation at ni, oF euanosine. methy| donor: sem (S-

Adenosyimethionine. enzyme : methyltransferase.

Functions +
*+ Prevents the ottack of 5! 2? exonuclease.
“GrientersdonORtotes
• The cap binds with cap-binding complex —> Helps in the attachment of mani to the
405 ribosome of ‘he translating macnnery —* initiation oF srarslation,
a. Poly A tail at the end. 40 ~ a00 Adenosine residues ore added to the Primary
transcript at the 2 end. ‘Enayme # Polyadenujate polymerase. Functions
• Prevents attack of 3! —> s! exonuclease.

• Stabiizes the mene,

• Poly A tail binds to PA (Poly A tail Binding protee) > Helps in the attachment of the
mena to the 405 subunit — Initiation of translation.

PTM - Removal of introns & splicing of exons _0035'54

Infrons + unwanted regiont/ non-coded to the proteins. Exons * They are coded to the
protens.
Biochemistry - V4.0 Marow 60-2022 47 vere 283
‘exons in human chromosome | MM t10n - intron pntton ‘se intron ton OG Sten sf ntron J
{GonorSte) ranch she >? en Gaerosne oF tron rescue) Gecepter end) steps
• SPRNIA + Protein —> snRNP doer ste 3) acceptor ste
earch ste denote) Seen cute? donor ste
atone
2! on frome tond A Spo, => a= phosphodiester bond
oe
CTR erat srctre ‘The non ie degraded
Spliceosome 2 components : + SeRNA: Transcribed by RNAP I, Uracil rich (uy ua, Ua)
example of the ribozyme. * SARNA + Proteins —> snRNP (small nuclear
Bibonucleoproteins). a/e/A seurps.
fin autowmenune disorder associated with snurps «6 SLE. + SARNP + exon — intron
junction oF primary transcript Grnant) —> spheeosome. Sel¥-splicing introns : *
Discovered by Thomas Cech. + coup Vintrons No Lariat Formation. + eroup N introns
Blochamisty “V4.0 - Marow 6.02022 284. Molecuar a7 Sosy ET Lariat structure is formed.
No ATP is required Sor both groups. 3 fiternate mRNA processing/ Differential RA
processing, The exons can be spiced in different alternative wus 50 that diferent protein
products can be produced Srom the same gene. + fterent Isoform of the same proten can be
generated. + membrane bound { secretory immunoglobulins.
RNA editing a sos + 00M mRNA undergoes hs mechanism. Bape: = mm wroren — ap
@marroweditionggolgs
oi Lye mare Tea & pn Oy ir
vara
Questions
I eA ens po ad mA of isones.
a. Poly A (RARE) codes Sor + Poly lysine. Poly: cugere
PolyU: Phenylalanine. Poy: Proline.
i 3. hARNR without intron : hn2NA of histones. i 4. Site of post transcriptional modification :
Nucleus. 5. S0RNR + Small nucleolar NA that is invoved in RXR
processing
Site of RAR processing : Nugleoks. So, called nucleolar RNA.
Biochemistry V4.0 Marow 60-2022 TRANSLATION
Introduction 00104 Oefevtion : Translation & the process in which protein s suntnesized
from RRA (men.
Site of translation : ReR (rough endoplasmic reticulum), free ribosome.
Genetic code & Representation of amino acids in DNA 00157
Genetic code : ‘Definition: the relationship between a. sequence of ohR and a Sequence of
amino acids in the corresponding polypeptide & called genetic code.
Cracking of genetic code was done by marshal Nirenberg and u &obind Mhurana.
cistron #5 tne smabest (AEH GORE LGRERL in codes for a pouypeptide chain.
Eukaryotic mRNA is monocistronic and prokaryote mx can be polyeistronic.
Representation of amino acids in DNA:
1 base represents | amino acid, hen only 4 amino acids can be represented.
the base sequence i of , then ko amino acids can be
represented.

the base sequence is of 3, then 4 amino acs can be

represented. .
(Triplet nucleotide sequence is sufSicient 4o represent the a0 amino acid).
Codon and salient features of genetic code 00849 Codon ¢
Definition : the triplet sequences present in mRxR representing specific amino acids are
called codons.
Blochemisty +40 Marow 60-2022
286 Molecular 48 Biology
‘stop codons : mong the codons, are stop codons erminator codons), namely UAA-Ochre,
UGA-Opal and, upe-Amber (UeA can be recoded to selenocysteine 4 URG can be recoded +o
pyrrolysine). Initiator codon + ALIG is the initiator codon; (met in eukaryotes § N-formyl
methionine in prokaryotes. amino acids are represented by a single codon - Aug-met
(methionine), uae-Trp (tryptophan). Amino acids are represented by the largest number of
codons (& codons) - Ser, Leu, Ary Salient features of genetic code : * qenetic code is
degenerate (redundant) ie, amino acids are represented by more than | codon. Degeneracy
of the codon mostly lies on the 3 base of the rplet sequence. * The genetic code is
unambiguous) a codon represents only a specie amino acid, © COR ERAS TORCH UPCALES:
* Genetic code is non — punctuated ie, there is n0 comma or semicolon in between. *
Genetic code is universal ie, 0 specific codon represents, a. specific amino acid, in all the
species. fa exception is mitochondrial ON.
Cellular machinery for protein synthesis and wobbling 00:16:50
Cellular machinery + Bilbosome = r2NA + specie proteins. + eukaryptic ricosomes contain
areains and 80 different proteins. i + drtypes of raMiin eukaryotes :a6S rRNA, 16S rRNA,
SES
RAI SS RNR [= ceetpie cane do ram 0 G37 divided into 405 and 60 subunits. + The 405
subunit has 185 r@NA and 20 proteins) the 6S, subunit has a85, 5.85, and SS rRNA and SO
proteins.
Biochemistry «V4.0: Marow 60 «2022
Transfer RNA (RNA) ©
Ao called soluble RAR (SAR.
The shape of the ten: the secondary structure
1s clover leaf-shaped, and the tertiary structure is
L-shaped.
Single tRNA contains 4-05 nucleotides.
42X08 contains the largest number ox unusual bases
(dinydrourac, pseudouridine, hypoxanthine).
12rR contains Thymine (pseudouridine arm contains. rieothymiding).
Arms of ean +
Amis acid attachment ste
Receptor arm: Has 3 unpaired nucleotides CCA of 3 ends binds the specific amino acid.
fntcodon arm : cleotide sequence complementary to he codon; binds 40 the codon n the
mRAR.
Harm / © - arm — recognizes the specific aminoacyl on synthetase enzyme (helps in
addition of the amino acid to the acceptor’s arm. Pseudouridne arm: Binds the charged
tRNA Ko the ribosome.
wobbling Codon Reticodon interaction at the 3rd base is nok very strict
For example : The antiodon loop of t2KRPhe (AA or ARE) can inkeract with codons. UUM or
ULC. in mRNA.
The reduction of the number of 21S & possible due 0 wobbling.
Biochemistry -V40- Marow 60-2022 288 Molecuiar a8 The phenomenon of wobbling was
put forward by watson and Crick,
Steps in translation & Energetics 2954
Steps in translation : 1. Charging of tena. a ination. “287 elongation. 4. Termination.
Charging of tenn: Ankicadon arm : UAC =» DHL arm recognizes the specific f2xR Synthetase
> specific aminoacy 4eNA synthetase. (RTP —> Ame) > methionine > amino acid &
attached to the acceptor’s arm. Identification of the initiator codons + The first Aue
sequence alter the marker sequence is defined as the start codon.
° MORATSFRYASEIRFTBISYSLeS + Shine Dalgarno sequence.
• marker sequence in eukaryotes : Kozak Consensus. sequence.
nition of protein synthesis volves 4 steps: . Disassembly of ribosomal units (xs elF3, eI, 05 >
Gos. 405 a. Formation of 805 intiation complex TP + Fa + met 12nR (Ternary comple) | +
40s subunit 435 preictiation complex [RNY 485 inition complex TP, ea +o |< 608 subunit
805 tiation complex
Biochem -V40- Marow 60-2022 48 mr emits gc Catalyzed by proteins called elongation
Factors (£F). Ws a multi-step process that includes. 1. Binding of aminoacy 2x8 to A-site
(requires EF-1, GTP. hydrolysis). a. Peptide bond formation usp Ear GS RS TS srs rare 3.
Translocation of the ribosome on the mRNA (requires.
£F-3, GTP huydoiys).
Termination + Complexed with RF-3 (releasing factor), &TP § peptic “transferase. +
Requires 2F § TP hydrolysis
energetics : 4 phosphates are required for | peptide bond formation. * Charging of {2A
“TERME AIR respites inorganic: phosphates), + FI ~ binding of #2xRad to A-site > GTR. +
era transhcation > 16TR.
No ATP s required for actual peptide bond formation.
Classification of RNA o0svan wn ] Founcagha] wrens — an [sea ne as rom= 5 a Cm won i —.
: = . mn 3 Circular RNR (circRARD : responsible for the regulation of gene opresin,
micro RNR (mene) :
Bochamaty -V40- Marow 80 2022 200 Woecuar 48 Bear TEs * Ws a small, non-coding,
single-stranded AR * Length: ato aa nt (rucleotide). + Discovered by Craig mellow { findrew
Fire. * It is endogenousiy generated from pre-mi2nR. Pri-menn + Pre-mi2aR (contains. Cap
+ Poky A tad | DROSHA DECRE Trimmed mi2nn | xportin-5 Trimmed mien | TReP dicer
nuclease sean | pegprause protens Loaded info RISC. complex (AR induced silencing copie.
+ mena © meghnemohertioonsnotes mi2xR © nvoived in the post-iranseriptional
regulation of gene expression. mRxR binds with the seed sequence present in the 3) UTE,
on the mRNA. 1 ts a perfect bose pairing : mx degraded. = 1% one or more mismatches are
present : Transition arrest. This process is called RA interference/@NAi = gene siencng. +
mnR © involved in the molecular pathogenesis of neoplasia. + Oncogenic mA + Oncomirs.
5208 (siencing a8 / small interfering QAR) :
} + 1s a small, non-coding, single-stranded RAR. i * Length a 40 3a nt (uieotde).
• mechanism of action : same as MENA

• tis exogenous in origin

Blochamisy -VA0 Marow 80-2022 Long non-coding nA Gnened + + 1s a product of xP 1 RA

polymerase 1D. + Function: Regulation of gene expression. + Different methods of gene
expression of IncZhR -
I. ene actaton : Facitates TF (ranseripton Facto) binding,
a Prevents gene transcription Inch + TF inhibits the binding of TFs to the promoter site.
Example + decoy RAR.
3 stone and DnA modification By directing
methyjases § acetyjases to the histones and RA. Thus, helps in epigenetic modification.
(@marrowedition6notes
Bochamaty -V40- Marow 60-2022
© 202 Molecular a9 Biooay
GENE EXPRESSION
Regulation of gene expression: Factors affect the expression of the genes : Genes express in
different waus in diferent situations.
£g; Insulin although present in the fiver they express only
the ponereas. Levels of expression of gene o0or20 on (gered)
§ Transerpton
men
Translation
Proteins.
ft Gene level | At Transcription level | A m2xA level
(post-transeription)
cone CTATYRUGATTOCS [eam sag ampificaton | repression Aternate Rx Gene i processing
rearrangement | Operon Concept (RNA epigenete (Ran interference) modification
Transposons.
ene suitching
Vousereeping/Constiutie gene inducibe gene I cpeccdotammtontiote. sot opressedata
B nbosalievelof expression constant rate. eg Hexokinase. expressed abter a stimulus. £3.
Slucokinase.
Blochamaty V4.0 Marow 60-2022
°C. EE >
Operon concept 000713
fn array of genes having a specif purpose. The concept was put forward by Francois Jacob §
Jacques monod. Lac operon
fn array of genes for metabolism of lactose in €. col
operator ste.
xR polymerase enzume binds fo the Promoter site. @marroweditionBnotes cegquatory/ |
structural/chcie gene:
coretiute gene a | Lac 1+ Secrete. Wa -y LER | vepressarteviotor | + + + | -gplactosdase
permease Thogaiactosdel Bind 40 operator + ‘ Sonscetjess) ii Lactose [ET % breakdown
enfry into the Function (Gactase) to cell. om— [glucose § lqoctose 8 polymerase cannot
move forward # the repressor/ inhibitor (active state) is bound to the operator site. i
Catabolte repression €2): § CAP/cRP : Catabolte actator proten/Catabokte repressor
protein. AP/cRP can be in an active state (oATP attached) or an inactive state (cAMP not
attached).
Blochemisy -V40- Marow 60-2022 204 Wocuar a9 Boor CaP/cRP 1 a positive reguitor of
the Lac operon.
• 1 CAP/CRP is active then the Lac operon siitched on. Glucose & a better source of
energy for Ecol (preferred Sued. Tuo conditions ¢
• Glucose s absent : ATP levels are high > CAP/C2P actwe.
• Glucose is present : cAMP levels are low > CAP/CRP active. In the presence of glucose
(catabolite), lac operon is sustched ofS (repression).
Scenario! sucose + | Seenar a: Gucose -1 | Seenar 3: Shcose + §
a [Pr [evant | Lacoperne Lao oper sched. | cae porn swtched wheat on a Coonedingtoc 11 |
qianed npiac 1108. | planed byce [SR meta notes [meterety [re cepresor/epivtor | cane evo
ae 0
rien [eho rae, | romeo ot go> coe ot avscoce (Bacon wo brs (Erar nasi
{i:4i
Isopropyl Thiogalactoside | Lactose analogue.
Gene amplification 02405
The proces by which the number of genes avadabe for transeription is nereased.
£2 Patients on methotrexate for a long time ~ increase in HF reductase by body > develop
resistance.
Biochem © =
Gene rearrangement 02631
v 1-1] [=
1 variable 2egon of rruncgbuin © VIDa V3 = unique variable chain.
(@marrowedition6notes OifSerent gene segments are brought together in different
combinations. fets at the OVA level eg, Immunoglobulin vasiakle chan is produced by joning
3 gene segments (v, 0, nd.) segments. V, 0, and J have different numbers of segments which
in variable combination produce a. different product.
Transposons w2902 mobile ORR sequences hat move from one location to another.
Decovered by Barbara McClintock.
Enzyme responsive for movement : Transposase.
Two Classes :
L1
Class 1 (90): Retroposons Class a (om: Transposons mobile OAR elements moving DNA
segment moves. with help mutation: insertion §
of an expressed QnA product. deletion.
Blochamy V4.0 Marow 60-2022 mas TH = -—mr
Epigenetics waza
Definition Reversible heritable chemical modification of RA or chromatin without altering
the nucleotide
sequence. Two Types. — ‘onA modification Histone modification
‘epigenome : Constellation of covalent modification of DNA § histone > Regulating gene
expression. DNA methylation I 003690 Taxes place of cytosine rescue wherever Cpe sands
present (seen in promoter region). Enzyme : OAR methyiransterase (mT. Metin donor +
Sam.
RPT APRIDRYES BRAS 105
5 2a a'~deoxyeytidine.
Decitabine.
methujotion of cytosine resues results in he ionbiton of gene expression — Favor
heterochromatin formation.
Histone modification 004005
Histone acetylation { deacetylation. Post-translational modification. Histone acetylation :
Positively charged histone & added with
negatively charged acety roe (erco0)
becreases the posite charge i ‘on becomes less condensed i Euchromati/Permissive
chromatin Increased expression of gene.
Blochamity VA Marow 80-2022 © on ET
Enum : Wistone Acetyjase (AT).
vistone deacetujation Increase in the positive charge by removing the negative charges. +
Increased condensation of fhe oN. i Hetero chromate. Decreased expression.
enayme :Hstone Deacetyfase (HOR. rugs ehbitng HORE + + vorinostat + afro acid
Functional consequences of histone modification + I. Histone acetyjation > fetation of gene
expression. a. vistone deacetyjaton =» Decrease gene expression. 5 vistone
phosphoryakomammmistddmioess ene @m& FETE Hionenotes 4. 1 phosphinfotion > Crate
cistern S. stone mefhation — increase/decrease gene expression. oP ibosujation > Ox
repalr: +. monoubiauitytion ~ nerease/deerease gene. expression ©. Sumoyjaton (Sumo :
Small ubiquitn-related moder) “> Chromatin condensation
Physiological & pathological ole of
gpeegel: + Regulation of gene expression i + Genomic imprinting Sight di¥Serences in
identical i alleles on paternal § maternal chromosomes. ° fgng + embryogeness.
Biochemistry + VA0- Marow 60-2022 298 Molecular 49 Beiony GEES Pathological + +
Fragie x syndrome : Fe -1 gene is hypermethyated. * cancer : oncogene is methylated >
prevents cancer. If the tumor suppressor gene is methyjated > causes concer.
• Pradter will syndrome § Angelman syndrome.
Molecular methods to detect epigenetic modification
00:55:21
1. methylation Specific Poe
2. ON Chromatin immunoprecipitation (ChiP).
Ge
YG —aeMadapBeeed to the chromatin with $ epagenetie modieaten
‘The antigen-antibody reaction takes place.
onlase added W yeprenrrenetianeriendiestromatin attached to the antibody-antigen
complex
DNA sequence taken out
Sanger’s sequencing microarray technique chip seq, (Chip technique) chiP chip. 3, BisulMte
sequencing +

Blochemisty “V4.0 + Marow 6.0 2022 HYBRIDIZATION TECHNIQUES

Heridization techniques D Biot techniques. microarray technique. Fluorescent in situ
hybridzation
lot techniques : + Southern elot, * Northern slot. * western got.
Southern Blot 00149
“1 bok technique invented. + Named after emsouthern (invented in 979). + Detects on +
asic principle + DNA - DNR hybridization. (@marrowedition6notes
PANINI ONE LE
| sot sien STIR SON
• sgprose gel lctrophoresss nn
n=)
nmr | rte toned
mpm buses
8 the membrane has Suorescence at a parteular locaton, then the sample & posite for
particular bacteria.
Bochemsty +40 Mow 60-2022 300 Molecular 50
a -—
Probe A singe stranded knoun dlgonuciestide. Labelled (radioacte/Fleuoroseent), uses: 0
microbiology (veal bacterial pathogens). sereen inborn errors. 2 on mutation stucdes. +
Large gene mutation
• Single gene mutation.
• Point mutation.
© Forensic medicine.
Northern blot 00m?
Detects eR. Principle + 2n- ON (COD hur dization. Separate eR
Lr @marroesikiegfnotes
[I
Teepe Lor
1 it detects Suorescence / radoactity then # © positive For viral enn.
con probe & used. 1 complementary to veal Rx. The val Rx s converted to ON using reverse
Hranserptose and cou probe. uses:
D To detect ex.
To detect gene expression.
Blochamisty - VA Marow 80-2022
tee 2
Western blot oota:t2
2/sfa.ienecunoblot detects protein,
W any Suorescence i detected, the sample is positive for the antigen
Principle :€Agfb interaction (immune reaction). Labeled antibody used heh is
semelementary ss antigen
Other blot techniques _ oos1716
• Southwestern biot : noun as Overlays blot. Detecting ONA protein interaction.

• pot bot/Sbot bit: Slotting with nitrocellulose membrane is not done. kis blotted to
several sts.

• 200 blot Used to study evolution,

Microarray technique _ oo:t948

‘Thousands of known oligonucleotides is impregnated on a.
slide.
uses:
• Genotyping,

• Gene sequencing,

Bochemisty V4.0 Marow 60 2022 302 Molecular 50 5 —

• @ene expression.
• Proteomics.
Classification D ona microarray a conn microarray. 2 rote microarray. 4 frray Comparative
Genomic Hybridsaton (Car,
DNA microarray 00224
y
SEE |wsaesics sed 558d
(© resipuner lignin déeode
In OAR microarray there are several sides with different wes. ach of these wells contain a
known digonuclotide. Inthe next step, an unknown oligonucleotide | added which is tagged
wth a Suorescent label 1 meets its complimentary pai # results in hybridisation. 1 not, t
gets washed ofS.
= “Wsarsslence i detected.
DNA microarray wasos
QOOOO sown conn |
OOOOO | urkroun ena labelled
l
OOBO0 computer agoritm
Bochamatry -VA0 Marow 60-2022 — 0 iimritie
cont comes sth tr eran on al
re ears mite rc ke ced
comme oli
Detect gene expression (especially oncogenes).
Protein
icroarray 02830
Foun b/g 0000 00
| Camere aig Grmoonsd (0000 00
Fluorescence
ohn tay reactors. ie peor
Toe uvren protien are added ober agg wih poten rset oc he cer SAFES Fact nie oa.
The Purousent ibe detected
Array comparative Genomic Hybridization 005253
Based on microarray technique. Genome chip : The entire genome of the organism &
deposited in the side.
a genomes are compared.
Bocnamatry -VA0- Marow 60-2022 304 Molecular 50 Bidogy
(ora care cone on
) 50m on ampitcaton
Sn json nase We sont Woo ne Normal Test
ellow Normal. areen : Gene deletion.
Red: Prgifigotion ofiasRon6notes
uses.
• Detects gene deletion

• Detects gene amplification.

• Detects geneti abnormalities in many diseases with unknown eto (autism, dusmorphic
Seatures, cancers).

• i camot detect balanced ranshcation 50 ee

Fluorescent in situ hybridization (FISH) 04853

@ FF
Dom fmm A eon ie
Perk hp
AX
px emma
epuorescent meroscopy
arroweditionénotes A technique to detect a specific genetic dement using
Suorescent labeled probes in a morphologically intact tissue.
muiteobur FISn/ metaphase FISH Spectral kar uohupng one ot metaphase.
S Fluorescent dues
mixed to form a3 distinct dye
£ach chromosome probe &
labelled wn a uniaue coir:
uses D Detects gene deletion (subtie microdeletior. 2 detects gene amplification
(exaggerated colour. 2 detects numerical abnormalities 306 Molecular Biology
50
4) Detects structural abnormalities (translocation).
9 Locate newly identified gene to is correct position.
Nuclear FiSw/interphase Fis one in interphase.
Rapid.
more sensitive than metaphase FISH.
For rapid result (prenatal sample,
tumor cel.
rousng cell 1s not needed. (arouing cells is needed in
metaphase FIs
Chromosome painting 00201
Labeling the chromosomes with different dues (not unique)
Guestion. what s the techniaue shour?
SpE Feesrston 8 doruieess FH = i
SE & siting Butter J. ose b AVRIL wei spree LLL 3 Ee oo | es or mise wee pats - Serge
Answer : Southern biot
RECOMBINANT DNA
Definition 00033
1 vvo amplification of technique used to get a clone of he.
desired onR fragment.
Restriction endonuclease wos Discovered by werner Arber. Breaks covalent bond 3' —» §
Phosphodiester bond hence i s a hydrolase (Class 2).
Ve resivcts the entry of phages into the bacterial cell 1s present side fhe bacteria. Types: +
Type Clik the sn ak a random ste. * Typed Cutt dsOR at the palindromic se. used in
moleculor bihgtechnigiéion 510105 ‘Discovered by Hamilton Smith and Daniel Nathan. Hs
olso Known os Molecular scissors. action + eCoRI is specific fo Palindromic site.
GATT soe mTTe Btey
CTTAR 6 coli CTT & ered end/
• ive end. Sticky end wth over hanging sequence

• Mpa ln Palindromic site — aTTie wal &TT MC | Guntend can (TTe cA TT

Blunt end - No overhanging Sequences

Restriction endonuclease s specific for bacterial palindromic: ste.
Cannot cut #s DNA due to site-specific methylases. GE te
Blochamisy v4.0 Marow 80-2022
51 308. Molecular 51 Biology
Vectors 01000 Vector is a molecule of DNA 10 which a. fragment of NA 10 be cloned can be
attached/inserted.
Properties + Vestors should have autonomous replication. + Atleast | restriction ste should
be present. + Atleast | geneman canfarantiaaresarstance.
Plasmids : These are circular dso present outside the nucleus. They ore extrachromosomal
OR.
8 ~ 10 copies are present in one bacterial cell
Funetion : To confer antibiotic resistance.
Plasmids con carry 0.01 ~ 10 Kop (on insert size) of foreign on
Plasmids have their origin of replication (or.
eocteriol ON©) 6rptes WR Plasma
roges
Proges: Viruses thot infect bacteria. Linear oR.
OMA insert size: 10 — 30 Hop.
a
cosmids Plasmids with cos ske/cos gene. Cos gene : It i required Sor packing phage ona
thas properties of plasmids + phages.
OAR insert size : 30 — SO Hop.
Artificial chromosomes Artifical created plasmids.
Biochamtry v4.0 Marow 80+ 2022 51 Sr lee
1# similar to a bacterial chromosome called as BAC. 1¥ similar to a phage chromosome
called os PAC. 1# similar to a yeast chromosome called as YRC. BAC 1 PAC - ONR insert size:
50 — 350 bp.
YRC 1 350 ~ 3000 bp.
WAC Human artificial chromosome.
Steps of recombinant DNA technology wo1s2a
Isolation of desired DNR Fragments. Selection of vestors.
Formation of chimeric ON or recombinant DAR. Isolation of clones.
Supthests of recombinant OVA:
oesred on _— Sie ——— ==
@| RE
sie | og (same on bot) formed.
‘Recombinant plasmid/chimeric DNA 4 multiple as the bacteria replicates Recombinases ~
a { ct ors [rarer i A att site. =r
Biochemistry v4.0. Marow 60-2022 310 Molecular 51
Beiogy
They are adjunct to restriction endonucleases. mechanism of action : Helps in the site-
specific insertion of the foreign DAR ky a process called. homologous recombination.
Foreign DAR inserted at LOx® site.
—
a |4fa
site specie esarton of Foreign On hy woriogns recast
Gene library 026.21
A collection of recombinant clones from a specific sources.
Depending on the source * + genomic irary, ‘The clone of the gene is obtained § arranged
‘The desired oxn Fragment is taken from the genome
onde SRO RYEttionsnotes
Disadvantages : dificult to replicate.
Q
“at
• eOM library The desired OMA fragment is taken from BAA
Reverse transcriptase Collect mana a cn
Advantages: + Less in size and no introns. + more informative and easy to replicate.
• can be used to study gene expression.
Biochemistry + v4.0 = Marrow 6.02022 POLYMERASE CHAIN REACTION
Introduction 00027
example of uses :
• From a scene of crime blood sample obtained. To detect the culprit using the ood
sample.

• To detect Covid 19 fection.

Ampification techniques | nevi. vitro. In-vitro amplification technique : = = Themakswkia:
Temperature of the reaction mixture varies, eg, PC, ligase chain reaction (LCR).
«isothermal cycling : Temperature of the reaction mixture doesk vary, e.g, NASBA (rucleic
acid sequence-based Rafi), BUA tence ranched oni technology.
Amplification technique
Target ampificators | robe/primer | Sgnal
. pee amplification |amplieation + QBrepicose. | techie.
Probe/primer amplification : Probe joined on target is ampifed.
Target meets probe Hybridization occurs — Attached with (Surcscent dyes — Signal
obtained — Amplified. PCR wor20 PCR 6 an in-vitro target amplification techie.
uses A
Invented by Farry © mulls, in 1969.
Ingenious technique.
Biochemistry v4.0 Mamow 80 2022
52
Act scn 312 Molecuiar 52 Biology
Prerequisites Sor PCR + sample OR (360/80 vn rate),
600m: absorbed by ORR
280 vm: absorbed by aromatic amino acids/proteins.
Numerator > Denominator | Pure sample.
adage Tosea
• ATP: For extension of primers.

• Cations mg" and K.

• Taq polymerase. VR polymerase enzyme lated from Thermus auatius.

Thermus aquaticus + Bacteria ving in hot geusers/mot sprog. ets ak high temperature.
Steps of PCR: + denaturation + aemealig, + extension. * oot arowedition6notes fonealing: +
Temperature requred: 54 To LO°C (ke cooing) + primers are requred Sor annealing
process. + primers can ‘be for ward. or reverse

on 2 ora {mmm | 1 mans pasar pcuaaass + Ser Be cxmer Sart cary Mrealng: a sets of eT ete
aitel oy, b ‘reacton mocture oarapene ERIM. imm@jns . ‘extension : = = ONR extended 3 +
seuoe _ SMEEEEIEEE Ss ihionsotes cree extention of target out done new eye started
denaturation (do not extend upto 3 minutes, ony intial de- naturation takes 3 minutes). fer
(expe of PCR a samples of NR ore produced Ge, al samples oF OND. after a™ cycle of PCR + 4
samples of OAR are produced Ge, aa: 4 samples of OND. Hence, after a”: (axn) samples of
ONA called as exponential amplification ip priate wi rogured tr 1 Peau Seonules: ‘petection
f
Blochomisty “v4 Marow 6.0 2022 314 Macular Biooay
tenon
52
Reverse transcriptase PCR (RT-PCR) 002508
Variant of pee. can ampify any ene. Enzyme used T polymerase (has both reverse
transcriptase + DNA polymerase actity). T polymerase enzyme 1 extracted from Thermus
Hhermaphius bacteria wal enn Lb corse varserptace
eT
l sep py Gide
[Io
aso
@ RGR notes
Undergges rex. Simplex PCR. muitilex PCR
One set of primer added at | multiple sets of primers
atime added ot a time.
One setof target oA | more han one target OR |
ampified ot a time. amplified at a tme
Time consuming Tene saveg ess fme
required)
more specif Less specific
Real time PCR & rRT-PCR cas
Real time pee :
Aso Known as quantitate Pee. ‘anpification and detection’ occurs smultaneousty.
Blochamatry v4.0: Marow 80 2022 =
Certain cejes (Suroscent), enzymes § probes added met tv od
Syee green
ar
FRET probe (Buorescence resonance energy rans-
fer).
A when target amplified Suroscenece emitted that wil be — Indicates amplification of
target and can quantify DNA § 238
RT-PCR (real time reverse transcriptase PCR) + Used for
cov, Applications of ICR ~ doswds + Forensic medicine example : DNR amplification of
culprit Som blood: + microbidlogy + @marroweditionénotes ‘Detection of RNA /ONA virus.
+ Study of mutation. + Cepeatummyte keen Aurec shvction Usually: 3s0nm Length wil be
increased hence more Surosence signals. detected. + Preiminany technique to other
molecular biobgieal teenies. Aon - mutated = 7 — = ts frst) frei
or saind — STIEIEY

1
No product
4
eps o detect mutation
Blochamisy- v4.0 -Marow 60-2022 316 Molecular Bickogy
toy
DNA SEQUENCING
Various techniques 00039
Maxam § bert sequencing, Sanger sequencing, Pyrosequencing,
Next generation sequencing,
maxam § Gilbert sequencing * His also called as chemical cleavage method. * 1 con only be
used for small fragments of ORR.
sanger's sequencing: + invented by Frederick Sanger: * His also known os controlled chain
termination method. ~..Ms an openious techniaue. mosEpepuias trstiiawes oles + old
standard technique for detection of mutation Sample OAR. ANT (deoxnucieotided. dideoxy
NT. Henou polymerase (on8P-1 from which §—>3" exonuclease activity is removed).
Biochemistry v4.0. Marow 60-2022 50 om ie Basic principle of Sanger's sequencing Fer xt
sss co a restore ie a unique set of single-stranded DNA molecules of varying
fre. eng AATP + ACTP + dGTP + ATP ox Porerace Tengte on oar dice ager
AAAL
ACTCAGATGCT ACTCAGATGCT ACTCAGATGCT ACTCAGATGCT
ACTCAGA ACTCAGATGC ACTCAGATG ~~ ACTCAGAT ACTA ATC ACTCAG AT A AC
[Seaweed rat din Surther,
@marroweditionénotes + The NA strands are then saccoted sira-asiamatad
gel electrophoresis, then read from top 4o botiom (3) to obtain the sequence.
ATP GdCTP ddGTP dTTP 3
7 < ‘ v x c i § ¢ T < ‘ o me 2 inn i i
Biochemistry v4.0 Mamow 60-2022 ep ET
Boor Pyro sequencing 001746
His a synthesis ype of approach of DNA sequencing, more sensitive method than Sanger's
technique.
The modified nucleotide is added with : * chemiluminescent signal * Bioluminescence.
Principle
ante T__ nucleotide
rom Tester ww wun uw repre S
. ~- @marrowedition 6g i» |
Puyogram
Biochem + 40+ Marow 6.0. 2022 oe ee
Next generation sequencing (NGS)
2120 1s a high throughput sequencing method. 1s a massive paralel sequencing Proce
Fragmented put mn = pls = mk end tapa —_— 1 a0 Todeg eb | adapior Laten s——= | Sze
selection === | fnpiieaton yet aneration [ Seeing Oifference between Sangers sequencing
NGS sequencing | next generation sequencing | [masse parce secure | fppromimates a bilan
base fpprosimatel mon basex/day | pda ene crsumiog me sai Com cow comma i
—
Biochemistry v4.0 Manow 60+ 2022 320 Molecular Bidogy
MUTATION
Definition and classification of mutations 000s
Defined as any permanent change in the nucleotide. sequence irrespective of functional
consequences. Occur in < 1% of population.
mutation epgenetes Polyrerphiom Ronormal permanent | Reversible chemical | ermal
variation change mrucleotide | modsicaton. rucleotide sequence. sequence No change in
ruckeotide sequence. Mutation in germ cell can be transmitted to next generation out nok
in somatic cell. M/C mutation : Point mutation.
a wr
once ston
ki
Stent mutation
0 chong i amen asd w -oae
soymes renee) the
“The mutation occurs due.
to yeneracy of codon.
oes he ana uy aus
44
rissense mutation : ‘Sutamic acd(ek) valine(val
Changs amen acd rat
coded.
Classification of missense mutation 00:07:48 “- ow
• conservatue: fi i
One amino acid s replaced by 6h fsparte acd (56) another with simiar characteristics. €oth
lu and fp are polar and acide.
• Non conservative ¢
Biochemistry v4.0 Marow 60-2022 One amino acid i replaced by another amino acid with
different characteristics. wh = Example + HbS mutation takes place | ' in" position of 8
gjobin chain. _ wl
lu: Reid, polar amino acid Val : Non polar.
Transition Ae——a @m: imeypgitionénotes ce—— 1 ce ST
Purine «Pure. Purine Puimdine Puimidine¢—> Pyrimidine Non sense mutation, insertion
and deletion 001537
J —— ra
• Codng codon replaced by cptens stop con
8° thalassemia: In the 8 globin + che coding codon? i replaced by UG Gop codord + Insertion
of a single nucleotide. + insertion of a single codon. + Insertion of a large gene. Deletion +
Deletion of a single nucleotide. + Deletion of a single codon. + Deletion of a large gene.
Aeon
Biochamisy - v4.0 Manow 60-2022 322 Molecular 55
se rT esha soos ro ter es
Frameshift mutation 0g17
Insertion or deletion that leads to garbled reading frame. Example : Tom AT PE. L Tom eae
te a Tom en TRE Tom eAT LoT Pre frnserton of 3 nucleotide 4. TOMPE ficietion of 3
nucleotide 1 insertion or deletion not in multiple of 3 : Frameshift mutation. 1 insertion or
deletion in multiple of 3 : No Srameshifk mutation.
eg: in cutie Boros at 508 position codon Sor Phe is deleted
Truncated mutation, run on polypeptide and
rinucleotide mutation ware @marrowedition6notes
Truncated mutation
i—a—3—4-s.. +30 => Stop codon
‘ Changed to stop codon.
Resulting polypeptide hos less amino acids than normal.
un on polypeptide
I—am34—5—e—1—E-9—10. + Stop codon changed to coding codon. Polypeptide derived
i longer than normal. example: Ho constant spring, Trinucleotide repeat mutaton
Amplification of a sequence of 3 boses. example : FM gene (C66 in fragile X syndrome.
Dynamic mutation : Number of repeats changes during gpmetogenesis.
30 > Stop codon
sso
Blochemisny v4.0. Manow 80 2022 55 Mutation 3
Mutation detection techniques 02610
Numerical or structural abnormalities in chromosomes can be detected by cytogenetic
analysis (FSH : Florescent in stu hybridization). onR sequencing an detect point mutations,
small deletions and insertions. oid standard method 10 screen for mutation but expensive.
angers technique | most widely used method for ORR sequencing Restriction fragment
length polymorphism 1 a mutation creates or abolshes a restriction site. Single strand
conformational polymorphism (55CF) : mutation results in conformational change and
oktered mobiity in electrophoresis enaturng gradient gel electrophoresis (Gs6e) : mutation
results in canforational shang apdatered mobity in electrophoresis. Oigonucleotide specific
huoridization (05H) Specific hibridization does not take placegeasER ak © mutation. RaRse
cleavage Used to detect small deletions and. insertiors. micro frray (NA chi) + qenotyping of
single nuclear polymorphism along woth mutation. Sequence alteration : PCR analyse.
—t
sanger's sequencing Restriction fogment Sele bose ke polymorphism primer Ge extension 1
Singje base primer extension ¢
fitter amplification primer created | base prior to target ONA etection of signals in
ampiieation alter hybridization
Biocnamatry 40 Marow 60+ 2022 324 Molecuiar 55 f=
To detect mutation that a¥Sects length of oR y Amplicon length Real time pCR multiple
ligation dependent
nase | probe ampisicaton (rem. more Surosence suggests nereased length
03246
Primer and stusSer sequence hupbridsed. Gap between probes gated by OuR igase.
(@marrowedition6notes fralysis 1 exons deleted : No hybridization —> No products for
amification 1 exons are duplicated Large amplicons. Copy number voriotions can be
detected using MLPA.
@ Describe mutation given below ? formal transeripk AUG ULL US: GAS met Pre Trp a
Mutant franser pt AUG ULC USE &h& 5 met phe Trp a
A. Sient mutation, Base Substitution, Transition
Biochem 40 Manow 60-2022 VITAMIN A.
Vitamins ee 0001:20
Definition: vitamins are organic compounds present in small amounts in Various 00d
substances, needed for the growth and maintenance of the body,
: heey Marea] ac orningighast | a vk
2. VR Crom 7-dehydro= 2. Pantothenic acid, chotestroD. 3 Botn,
shin psunlight Uv 6 rays G4BD- | (all are synthesised rom the
aso) ntestinal Sora).
Classieation of vitamins +
Fat soluble : Vitamins 8, © ¥5 Woter soluble : 8-complex vitamins § vitomin ©
eco PARC UTTIONENOTOS
LG Thames. Foie acd @). 0, Pyrdoxne 2.8, !Ribofavn, cobolamin 3 @, 'Mocn
(Properties [Fat souibie vtamins | water soible vitamins
[recorption [needs chuyjomseren [Doesnt need
Se ee ee
esis? a lneeedpesime lures |
Tonety (present event (exception = Nac, 10.
Fncten | vareus function [ets a8 coeraue Groat)
(ars can aot os
[core
Biochemistry * v4.0 + Marrow 8.0» 2022
ew spsce
326 vtamins
55
Vitamin A 001014 Types. * Pro-vitamin A: mainly Srom plant sources; known os.
carotenoids.
• Pre-Sormed vitamin A mainky from animal sources; colectvely called as retinoids.
The ring structure present in vit f none ring + oprend
unt
B carotene has a ionone ring,
Cartencs,
(rete Prowama®
metabolism of vitamin A: eet carotene & converted
oko retinol in the intestine,
by the help of dowgenase =p
ERE 1% Monenotes
Carried.
— retin ester (palmitate) the perisiusaidal cels (to =)
[ Carried to target stes the plasmas trimolecuiar complex bound 40 26 and. wranstyretn
2etioids :
Al compounds related 4o retinol are called retinoids. 3 forms of retinoids : + Getinal +l
trans retinal and, i-cs retinal (present in rhodopsin); required mainly Sor vision. + Retinoic
acid : Al rans retinoic acid, 9-cis retinoic acid {
13-cis retinoic acid ; required for growth, morphogenesis and cellular ditSerentiation.
Blochamaty + v4.0 + Marow 80 + 2022 7
• Retinol Important for reproduction. Carotenoids
Carotenoids are provtamins.
major source are the plants.
most prevalent carotenoid with Pro-vitamin A activity B carotene.
Non-provtamin & carotenoids
• Lutein § zeaxanthin + used in @x of macular aton.

• Lycopene : Used in Rx of prostate cancer. Vitamin fo veion ¢ Ici retinal + opsin Sorms
important visual pigment, rhodopsin. wald's visual cycle Rhodopsin (1-¢is_y git
(Photon)

ronal pee). impulse ea) 1c retiat TPATTDHEGRGRALGS won + I go Alcohol NAD" RD"
wiver) Nol SOR cohen yea et
Series of isomerisation occurs in conversion 40 I-cis retinal & protein coupled receptor :
GPCR.
active spc MEEBO Ly cong
Active & protein s called as Transducin
Inactive Phosphodesterase "" acte prosphoc-
esterose. comp active Phosphodiesterase. , gee i a t Vupecpolarisaton 1 generation of nerve
impulse
Bccnamatry + v4.0 + Marow 80 + 2022 328 vitamins
55
Regiation of gene expression by vitamin A W acts via retioi ac receptors (2A). lgand &
retinor acid
• All trans-ra. and 9 cis—ra. are transported to the nucleus of the cell bound to
cytoplasmic retinoic acid binding proteins.

• within the nucleus, all-trans ra, binds 4o retinoic acid receptorsiror) and 9-cis ra. binds
to retinoic acid x receptors).

• Rar and rxr form rar/mar heterodimers, which bind to regulatory regions of the
chromosome gene called retinoic

acd response ElemetsprR Hence Vit Af vik 0 are cioted. :

interr
Vitamin A have steroid bormane fise function. Binding oF all-trans. ra and 9-ci ra fo rar and
ox respecte allows. the complex to regulate he rate of gene transcription.
Functions of Vita + + maintenance of normal mucosa. fskin. + Regulation of gene
expression. + Normal reproduction. + thas antioxidant properties (especially f carotene) * i
also has photoprotective properties.
Biochemistry + v4.0 + Marow 6.0 + 2022
Deficiencies of vitamin A 02024
Exes: ll manifestations of eye together called as Xerophthalmia. + Loss of sensitivity
towards the green lght (eartest manestaton). * Night bindness / nuytkopia. + Conjunctal
and corneal xerosis(dryness). + tots spots (due to conunctol keratinzation. + Corneal ulcer
and keratomalacia (corneal degeneration. Skin § mucosa Follculor
huperieratosi/phrynodermaload skin. Squamous metaplasia in the mucus secreting
epthelum
vera) gon phggen
" atosis and purple and is B marrohsdiipnanoies
Toads skin
Squamous metaplasia in the mucus secreting epithelium
ramaticimprovement after montis of treatment with Vitamin & can be expected.
Uses of vitamin A & vitamin A toxicity oss
most common cause of preventable blindness.
Uses of Vitamin A n treatment Beta carotene is used in cutaneous porphyio. fl rans retinoi
acid are used in promyelocytic leukaemia as a differentiation therapy. 13-cis retinoic acid
Gsotretinoi) i used n cystic acne, peoriass, and ehidhood neuroblastoma.
Biochermsty + 40 + Marow 60 + 2022
57 wenn as
330. Vitamins 55
Vitamin & toxicity/Hypervitaminosis + ‘Commonly seen in Arctic explorers (due to intake
of polar bear ver). Organelie damaged : Lysosomes. ‘manifestations of acute toxicity +
Pseudotumer cerebri. + exfolative dermatitis, + Hepatomegaty + Hyperipidemia,
manifestations of chronic toxicity + Nauseo, restlessness, weight loss, anorexia, + ony
exostoses. + Hepatomegaly § cirrhosis. In pregnancy : koh eon reraragenie. Retinoic acid
Savours osteoclastic activity causing demineralization of the bone, hence increases the risk
oF Fracture.
RDA, Soars ACER ESOS of Vitamin A
Recommended dietary allowance of vit Chidren (-eyears) : 400mea/day, men :
eoomeg/dou women : eoomeg/day, Pregnancy : Boomea/day. Lactation : 95omeq/day,
Sources of vita * Primal sources : Egg, liver, cheese and other mits products. + Pldnt sources
: carrot, green leafy vegetables, mango, Papaya, pumpin, * Halibut iver ol: Richest source. *
cod iver ol: a richest source. * carrots Richest plant source. Assay of vit. + bark adaptation
time. Serum vitamin A by Carr § Price reaction.
lochemisty + v4.0 + Marow 6.0 + 2022 VITAMIN D, E,K
VitaminD cB
Source sun ot a rue vitamin sine it & endogenoushy sypthesczed.
aroup of sterols having a hormone lie function. Tuo sources Plant source : Commercially
avalable vitamin. ergpcaleferal (tamin 0). Fung rapt. fnnal source : Cholecaleiferol kam 0).
endogenous sunthesis of vitamin © 5 we @s0-35 0m 7 denydrocholesterol Dietary Sources
d toad Cholecalcferol (03)
wr CT ores
1* step takes place in skin + ~dehydrocholesterol — Cholecalciterdl. Cholecaleifersi i
transported in the blood by 0-inding
protein (0, gobutid. a step takes place in the ler and 3" step in the Kidney.
eogealy most potent
Blochamatry 4.0 Marow 60 2022
hearse 332 vtamins 56
at hupdrossfase © actated by Parathyrod hormone and o decrease phosphate level
1¥ the body does not need vitamin O,
25 diuydrony Choecaleferol Calero is converted nto 24,25 diydrony cholecakeferdl
CalcetroD.
Functions of vitamin © + + Regulation of calcium * immunomodulatory action
Regulation of Calcium 00711
Primary function of vitamin is fo requiate Ca" and PO," Rese, rape do bSED Orgons |
Intestine, Kidney, and bone. * 135 (OW), Increases caleum and phosphorus. + PTW:
Increases calcium and decreases phosphorus.
Acton on Kneys + In distal ubules + 435 (04,0 — Increases level of Colbindin 38% and
TRPV 5 i Favours re-absorption of calcium and phosphate i i Increased serum calcium and
phosphate: Parathyroid hormone : Favour re-absorption of calcium and increases
excretion/decreases. reabsorption of phosphates Ehrosphatur) Increases serum caleum
and decreases serum phosphate. fiction on Bone +35 (OF), and PTH Favour RANK figand.
Biochamisin v4.0. Marow 60-2022
S6 er
present on osteoblast.
Rank ligand + Rank ligand receptor on pro-osteoclast leads to Sarmation af. Made.
asteaciast. +
osteoclast —> pemineralization of bone. 4
Fovour bone resorption (Favour serum calcium level.
• Vitamin © also causes mineralization of bone : when serum calcium low —> Bone
resorption. when serum level is enough — Bone mineralization. mineralization by
stimulating the osteoblast
• Increase osteocalcin level (eakium binding protei)
• ‘Deposit calcium in the osteoid matrix ond. epiphyseal carttage. Immunomodulatory
action of vitamin DC10(05 001405
‘exhibited in tuberculosis.
Fils muobd 1, as OH), 0 —» Catheliedin (A
ES rons meres tr. Vitamin © deficiency is a predisposing factor for Covid-19 and preserioed
os preventwe. Vitamin © The holy grad of cancer medicine. + a5 OH D level <0 ng/ml s
associated with an increase. in the incidence of Colon cancer and breast cancer.
Biccnamstry v4.0 Manow 60 2022 oo4 wrens ==
• Vitamin 0's protective of pre-diabetes and metabolic syndrome. Assay of vitamin
• Normal level of as cholecaleiferol a0 ~ 100 ng/ml
• Serum osteocalcin s measured to assess the Vitamin D status of the body nrekets,
é
Biochemical changes Vitamin © deficiency
. 4 SeRFRaLA UERIMCEAWUAFdecreased serum
phosphate (serum Ca can be normal also). Secondary hyperparathyoidism
Increase at hydroxylation 4 18 dinydroxy cholecaleiFerol 4 Increase serum calcium level
but serum phosphorus, level is decreased
molecular defects :
} Vitamin dependant Rickets | Gene encoding Renal i Type Pseudo-vitamind | -hyroxuyase
resistant rickets) 7 __| vitamin dependant Rickets | vitamin 0 receptor tape atta vtemind |
resistant rickets) l
Biochemistry + v4.0» Marow 6.0 +2022
X-linked hypophosphatemie | mutation in PHEX gene | riexets ap]: me. —* increased FeF
Autosomal dominant
hypophosphatemic rickets
Autosomal recess.
hupophosphatemic
Toxicity of vitamin D 02800
Adults > 4000 I and in infants > 3000 manifestations + Caleinosis: contraction of bood
vessel (increased Hood pressure) * metastatic caleifeation. The richest dietary source of
vitamin ©: Halibut ver oi. rll chetary source (anima : Fish
eon © Chidren10 mega AGP MA FEE IGOB ERoouse ones are developing). * Adis + meg/day
: 200 * Pregnancy : 10 meg/day : 400 Vitamin E 00:31:05
Aso known as Apha~—tocopherol (Bdigeally most potent). Steredisomers of tocapherdl
Ring structure in vitamin € + Chromane/ Toca! rng, most potent naturally occurring
antioxdant. Lipophile chain breaking antioxidant : Prevent pid perosidation. .
«Protects the Low from oxidation
• Protects the PUFA in the membranes Vitamin € deficiency :

• Axonal degeneration

• perpheral neuropatry

• spinocerebelar ataxia

• Moematc anaemia. (26C membrane integrity is bs)

Blochamty v4.0 Marow 60 2022 336 Vitamins
• Eyes: Pigmented retinopathy, ophthalmoplegia. Napogris
vitamin € used as treatment: * Retrolental Abroplasia. + Intermittent claudication. +
eronchopulmonary dysplasia. + Intraventricular haemorrhage of prematurity, slow ageing,
208 males :tomg/day, Females: emg/dayy Preananey :1oma/doy, Lactation: lamg/day,
Vitamin € toxeity manifestation +
interfere with platelet aggregation Interfere with vitamin @marrowedition6notes
Selenium decreases the requirement of vitamin &
Vitamin K 003626
6 stands for @erman word Hoagulation. Naphthoquinone derivative. Three forms +
Vitamin, Phylloquinone (dietary sourced.
4. vitamin 6: menaguirone Bacteria! Sore).

5. Vitamin 6, menadione (Synthetic water-soluble). Functions + Post-translational y

carboxylation of glutamic acid in

• Factor a.

• Factor 7,9, 10.

• Protein ¢ and Protein s.

• Nephrocalcin.

• Osteocalcin

• growth arrest-specife gene product (gas.

Biochemistry «v4.0 Marow 60-2022 — ops

i i SNR A
warkarn and Dieumarol (structural anlogue).
Garmrma Pten vitamin 1 - dependent Corbortfated
i
~7 rns ax” Ox” Vitamin 1 epoxde o oN Vitamin ne se T Ee
ann
(Quinone)
Vitamin deficiency @marroweditionbnotes Increased prothrombin time and bieedng time.
Bleeding maniestation.
Newborns are suscepisle for deficiency of vitamin becouse:
• Decreased fo stores.
(Sat needed for transport of Vitamin 1) + Steriity obgos ~
• Breast mik a poor source.

• Less placental transport

• immaturity of ngs.

Vitamin ts toxicity : Hagmolyss +

etic { aoc wou arcs i
Blochamisy v4.0 Marow 60-2022 pore 9 -_ HEMATOPOIETIC VITAMINS
Hematopoietic vitamins : Vitamin &, and 8 Vitamin, / Fol acd: mainky plant orig, rich in
green leafy vegetables (Folum : Leaves).
VHamine, / Cobalamin: Animal origin.
Pure vegans develop Vitamin ©, deficiency.
Folicacid - 00350 Retive form Tetra hydro folate (THER. Carrier of | carbon group. One
carbon group and their metabolism: “The | carbon grougs are as Follows :
methy (cH).
riBuediion 6rotes
Formimino (-CHen, 0 these | Cash ous ave reversile cept : Methujene
Tito mein eA vale em oe orc ebenme worm cine il NO el TE mocgice 5 TIP spthesis (OUR
Syphesid Foamimino— Metheny 7 men hice Metdine ~N
most important entry point +o 1 carbon metabolism is serine o glycine conversion.
‘enzyme involved in this conversion is serine hydroxy metry transferase.
r—
Blochamisin v4.0 Mamow 80 2022 57 Homa en Folate trap may lead to decrease in THER,
thus carrier For | Carbon group decreases. THER deficiency leads fo nuclear maturation
defect.
Folic acid deficiency uw manifestations © Decreased DR synthesis (decrease in
TMP/Purine rebate erie
methionine
mar awl
vomocystene
Neural tube defects : Spina. bifida, anencephaly,
mo TRE wattobe Lg HH on
itself, or atieast during pregnancy. Homocysteinemia : Recumulation of homocysteine,
HomocustnuricoTuBy skeMRBg
meth] donor(SATY decreases : Epigenetic modication affected and leads to cancer.
Normal diet i rich in folate yet deficiency possible as Foujpolygutamate(Chemical form of
Fol acid) +
‘Sensitive to heat, on heating green leaves and vegetobies for 5 4010 minutes destroy 95%
of Fol acid.
Biochemical assessment + Serum folate and Red cell folate. + Wetidine load test :
Formmien THER
™ Fle (Formiming glutamic acid) —— au Flot s accumulated § excreted in urine.
Blochamisy v4.0 Marow 60 2022 340 viamins
57
• AICAR (Amino imidazole Carboxamide Ribose 5 Phosphate) excreted in urine (Purine
Synthese).
• Serum homocysteine.
• Peripheral smear : macrocytes. Tear drop cel. Vyper segmented neutrophis.
Arisopoikioeytosis.
methotrexate(it folate) Causes folate deficiency, Folic acid/ S Formy THER : Treatment Sor
folate deficiency
caused by anti folate drugs.
Leusovorin + Racemic mixture of Rolie acid can alo be used. Called os leucovorin rescue.
Vitamin B,, / cobalamin 002553
4 puyol rings with cobal(a351) n the centre.
msc farton oR Cost 0n 610105
Absorption: mouth (passive form 9) , leu (ative form 99%. Haptocorin : Cobalamin binding
protein.
From dS Lmoutn : Ee SDN opie prone snr)
n i Sh the protein of cobalamin
Parietal cel (cobalamin + haptocorra ) trie et) w
a digest haptocorrn "cobalam + intrinsic facto of castle W. leur + Hos receptor for intrinsic
factor — Cubiin.
Cubilin binds with IF and cobalamin token up into feal cals §
Blochamisny v4.0. Marow 60-2022 57 Homa iam
transported in blood using TCa TC 1+ tagret organs.
Vitamin B,, / transport 0031.00
Transeobalamin (Transporter protein) Tuo types. Transeobalamn 1 For cobalamin analogs.
Transcobalamin a : main carrier protein.
Active Forms of vitamin 8 (Cobamide enzymes) + edenosye,
• methye,
Penog.@. HAUS AISARYETS 0 © + methyl malonyl Con mutase. + Leucine amiomutase (sent
in humand).
Propiny Caf enters. gluconeogenesis pathy through hi. Leth matory Cah #denosy 8, metry)
on oxfamidaseino les
Succ con
meth 6, plays coenzume roe n+ methionine synthase/Homocysteine methy transferase.
Homocpsine| re a methonine ‘Deficiency of Vitamin, deficiency (Strict vegan. malabsorption
: Autoimmune gastritis GF destruction) pernicious anaemia. Gostric causes. Intestinal
causes leu site of cobalamine absorption. + Veal resection. + Cronnls dsease. «stagnant loop
sycrome. Fish tape worm (Diphyiobothrium latur)
Biochemistry +40 arow 60 +2022 342 vitamins 57
Manifestations oars
megaloblastic anaemia. (more common in THFA deficiency. meth THER cannot be cnverted
to THEA in vitamin 613 deficiency.
THFA deficiency causes defective nuclear maturation. Homocusteinema.
meth molonie ackduria.
Serum methyl malonate + Di Serentiates megaloblastc anemia. due fo Sol acid deficiency or
Vitamin 6, deficiency.
Neurdlogieal manifestations in vitamin 8, deficiency meth molony Cah mutase affected.
fecumulation of L methyl malony Co and Propionyl Col. +
Proponi acid level increases as a result 4
Ronorosl fatty aed aRsvEatin to the neuronal pid i
uel breakdown occurs. 4 manifests as SSH SHE ERféneration.
Assessment of vitamin B,, deficiency oouzr0
• Serum cobalamin.
• Serum homoaystene.
• urine homocysteine.
• Serum methymalonoic acid: unique for Vitamin 83 aetiology.
• sehiling test i + peripheral smear. i + one marrow eytdogy. Q uh megaloblastic anaemia
in foie acid deficiency ? Decrease i TFA — decrease in TMP — Affect oA
Synthesis.
Bochamatry v4.0 Marow 60 2022 a virile Sere @. why strict vegans develop vitamin @ia
deficiency ? ecciles hes Ack secede GOK
@ why neurological complications only in amin 61a deReieney ?
ecumulation oF L methyl malony CA and Propiony) CoA. ‘
Propionic acid level increases as a result sora ety cates tosei reptnieatinn manifests 0s Sub
sss ess degeneration.
why isi essential fo exchide vitamin @ 12 deficiency as the cause of megoloblastic anaemia?
Ana case of megaloblastie anemia due to vitamin 6, deficiency, Fits not doarsend
ReSMRSBR RSHERLS 9 Place of 61a, tt leads to agoravation of neurological symptoms.
As folic acid supplemented used for conversion on methyl
‘THEA to THFA and 612 will be excessively used in conversion. 21a store further depletes.
rs
‘Blochematry v4.0 = Maow 6.0» 2022 344 Vitamins
ENERGY RELEASING VITAMINS
Vitamin B1 000203
Required for carbohydrate metabolism. The sources of thiamine are +
• Pleurone layer (most importand : Between the white
and brown layer of cereals.
• Unpolished rice, Whole wheat Sour and Yeast. The aleurone layer is lost when the rice
is polished. Parboling ic the prefered method of cooking rice to prevent this. Coenzyme
role fective form : Thiamine pyrophosphate (TPP) or Thiamine diphosphate. “Thymine
pyrophosphate acts as a. coenxyme in the Following, enzymes :
Pyruvate
ao niger apes
3, Granched chain ketoacid dehydrogenase.
4. Transketolase (non oxidative phase of He).
Deficiency of Vitamin B1 00106:18
Common in alcoholics. Alcohol decreases absorption of thiamine.
~ lek Bari dare Associated symptoms are §
LL High output cardiac failure. a. oyeproea.
3. Cardomegaly

4. Per gheral edema.

S. Pulmonary edema.
Case +A chronic alcoholic presented with pain and paresthe- sia. of wer limbs , with
cramps. Erythrocytes transeketolate activity ¢ reduced, which & the vitamin deficient ?
fosiver + Thiamine deficiency causing ary beri beri
lochemisty «V4.0 Marrow 60 = 2022 58 Eno iam ory beri geri: Peripheral nervous sustem
affected. Symmetric motor and Sensory neuropathy Pa, paresthesia, oss of reflexes
especially in lower limbs. Muscle eramps, Muscle atrophy(Severe cases).
wernieke's encephalopathy Case : chronic alchohol presented with prose of eyelid ,
confusion, nystagmus. Diagnosis : Wernickels encephalopathy. Central nervous sustem
fected. tren en TAGS. Opnthabmoplegia(Ptose). Truncal ataxia. contusion.
wlernicke's KorsaKo?S syndrome werricke's encephalopathy ‘associated with memory loss,
(Dementio) and confabulatory
pry eee: @marrowedition6notes Symptoms aggravated in Transketolase deficiency.
Triasmine phosphoryates choride channels in nervous system,
activates nerve conduction.
Biochemical assessment of amine deficiency eryiivocute Transketolase.
urnasy thamine excreton.
Required day allowance (20M 11-15 mg/day No reported toxicity.
Riboflavin/ Vitamin B2 _on1ase
Questions uring phototherapy which vitamin s supplemented ? i Answer : RiboSlavin.
{ When we take B-complex vitamins , what the reason for
discoloration # urine 7 Answer : Riboflavin.
Biochemistry + V4.0 Marow 60-2022 346 vitamins
58
whieh vitamin s a redox vitamin? Answer : iboSlavin.
Cooking Sood wil not destroy this vitamin? Ansier : Riboflavin.
Pigmented wtaminuarburg yellow enzyme) : Gives urine yellow colour.
83 @iboTlavi) and 83 (ack are redox vitamins. Supplemented during phototherapy (Light
sensitive viamie). Heat resistant vitamin
Coenzyme roe @ibotavn fictive form + Flavin Adenine Dinucleotide(FAD), Flavin mono
NucleotideFmnD. Fw coenzyme role Complex in Electron transport chain(uRoH
dehydrogenase). L-Amino eg pieMfedition6notes FAD coenzyme is associated with he
following enzymes © fy) Con dehydrogenase. a. Succinate dehydrogenase (Complex I in
ETC). 5 © finoocid owdase. 4. Xanthine oxdase. S. Al mult enzyme complexes.
Riboflavin deficiency 001956
Intilly asymptomatic.
Cheosis Pollr in angles of mouth. Thering ond maceration of eptheium.
Fisuring extends radially to sin.
Blochamisty - VA Marow 60-2022 * rt -— Be erin oes =
el
eyes + heats + Conpunctts. + Protophabia. f + Lacrimation, ¢ + Comeal vascularisation N
(Chasacterstic Sndng.
Other Features
Seborrheic dermatitis
Normocytie normochromic anemia.
Biochemical assessment of Risofavin eryfivocute Glutathione reductase after giving FAD in
vitro. urinary excretion of ribof@ifarroweditionBnotes
20815 mo/dayy
No reported toxeity, Niacin / Nicotinic Acid / Vitamin B3 002600
fn endogenously synthesised vitamin from an amino acid, Nagin actve form ore,
ARO Gicotinamide adenine
dinucleotide).
POP (Nicotinamide adenine
deucleotide phosphate).
symmetrical eryfhmotous seloy lesions. on the sun exposed.
Blochemisty V4.0 Marow 60-2022 348 Vitamins 58
“Casal Necklace” due to protosensitie dermatitis.
The lihenifed lesions on both the hand is known os Gantiet of pellagra.
NRO’ Coenzyme role *
every dehydrogenase requires NAD, Except enzymes hat reauire Fan Or NADP,
NROP' and KROPH coenzyme role AROP" generating reactions :
1 tuo enaumes of oxidative phase of Hexose monophosphate Pothuay :
• Gucose ~~ Phosphate dehydrogenase.

• to- Prospho gluconate dehydrogenase.

• Cufoplasmic ocitrate dehydrogenase.

«malic enzymes.
NAP requiring enzymes :
All reductase recuire NADPH.
oy mhenisan ia psidsuthess
« etoacy reductose : For fatty acid sunthesis.
= HI COR reductase : For cholesterol synthess.
«Glutathione reductase | Free radical scavenging
• Riconucieotide reductase | Conversion of Risonucleotide to Deoxy ribonucieotide.
«Folate reductose.
Niacin Deficiencies | Initially present with vague symptoms.
i + epressive paughose + eth Gevanced cased). + Casals necklace develops i sun exposed
areas
Syrneirieal,
Blochamiy V4.0 Marow 60-2022 0 wr ier pe Pellagra lke symptoms : Hartnup's disease *
Due to decreased tryptophan. Carcinoid syndrome: Tryptophan used for serotonin
production. ©, deficiency inhibits kynureninase. maize/corm diet : Niacin in bound form.
SorghumCiar) : gh leucine (hibits @ PRTase).
Prostaglandin medated Cutaneous Sushing. most fatal: Fulminant hepatitis.
ahucose tolerance.
Vper uricemia.
ostric rtaton.
macular edema.
Liver enzymes are increased
© fulminant hepatitis.
Treatment © @marroweditionénotes Premedication with spin.
Laroppront(Prostaglondin 0, iibitor). Nain: Lipid modifier drug, Decreases Triguycerdes,
increases Hot, Decreases LoL.
Pantothenic acid/Vitamin BS 03502 eta alanine is present in Pantothenic acd which s nturn
present in Con Endogenous sunthesised in intestinal Sora. Coenzyme role: Present
Coenzyme A Required for: i + Succiny) Con and Reety) CoRCTCA cycle) i + Has
CorlCrolesterol and ketone body systhess, Leucine catabolism).
• feu carrier proten(Fatty acid synthase comple)
Biochemistry +40: Marow 80-2022 350 vitamins
Deficiency manifestation : Gopalan's burning Sook syndrome! nusritional mela.
Biotin / Vitamin H/ Vitamin B7 oasis Case +A young adult who Sond of eating rauw eqqs
dauky developed erythematous rash around nose, eyes and mouth. which enzyme actwty
affected in this person 7
Ans + Rau egg contains avelin which inhibits biotin.
Active form : Carboxy biocytin,
Coenzyme role: Biotin dependent carboxyjation reaction. * Propony Con carbojase. *
Pyruvate carboxjase. + Roety) coenzyme A carborjase.
60th Independent carboajation Dc is
• molic enzyme.
• AiR carboxyase For purine synthesis.
• Carbomyphosphase synthetase land a
sot deficiency mental changes(Depression, Hallucination).
Scaling, seborrheic and erythematous rash. ounanasewEeases atte form of biote deficiency :
Leiners disease.
Other uses Ei0tin afSinty toward avidin is very strong, Streptavidin : Capable of binding 40
4 bite, used in laboratory investigations.
Blochomisy +40 Marrow 80+ 2022 VITAMIN B6 AND VITAMIN C
Vitamin B6 00123
Retive Form and Co - enzyme role Pyridoxine (ring structure). Aotive form: Puyidoral
phosphate (ALE). Needed for Amino acid metabolism. Transamination reaction: fet AT
alyojate Alanine amino transferase
b
resultggnigvbaelition notes
a Amino acid decarboxation |
vistidne °F; Histamine S-wydroagtryptophan °F, s-vydroytruptamine (erotonied Sutamate
= hen co; Transsulfuration, tryptophan metabolism,
heme synthesis and glycogenolysis anos?
2) Transsulfuration vameigteine + serv op | Cigtaionine beta sce
Homoserine + custie Transfer of SH group from homocysteine fo serine.
Biochemistry V4.0 Marow 60-2022 352 viamins
wou
4. Tryptophan metabolism : 2-Hygdrony Kyrurenine
PLP | ynureninase -hydroyantiraniate
Fetogenic fate NADPH metabolism
In case of PLP deficiency {
3-Hydronfyurenine > Xanthurenic acid leads to pelagra.
S. eme synthesis : Sucsing Co A+ eyeing. PP | a systhose ie veme In PLP deficiency :
mierocyic anemia. (@marroweditionénotes
Rate lmiting enzyme : Glycogen phosphoryjase Ste : Liver and muscle (80 4 stored)
Manifestations of Vitamin B6 deficiency 01200
Neurdlogjeal Peripheral neuropathy. Personality changes (depression § confusion).
conwision.
mierocutic hyposhromic anemia.
pelagra.
Urinary analytes excreted in PLP deficiency ¢ . ine.
• xanthurenic acd
• Oxalate (causing oxaluria)
Blochamisy -V40- Marow 60-2022
on EET
PLP and hormone dependent concer Vitamin 8, : Inhibit binding of Hormone receptor
complex to hormone receptor elements.
Deficiency of vitamin 8, leads to enhanced binding
action of hormone. Toxicity : Sensory neuropathy
Biochemistry assay
enyivocute transaminase. Tryptophan load fest measurement of PLP in blood.
on: - a mg/day
Vitamin C 017.2
water soluble. James Lid used lemon for treatment of sunny
most armas sypinesize vitamin C from. gucose.
Humans cannot sypthesize vitamin C due to absence of ‘L Guborolactone owdase’
Functions : ‘udroxyjase’ LCC comgrson ange aIneg uorases. copa B rouse Pept ghycne
hukoajase
354 Viamins 59
a. Coenzyme for @ ketogutarate fied iron containing
enzyme : Proline and lysy hydroajase. Decreased hydroxylation of proline lysine
defective colagen > bleeding manifestation
Anemia Ferrireductase (need vitamin ©)
Fe" Fe (Ferrous form) (absorbed from intestine)
Hence, vitamin C deficiency causes anemia.
Clinical manifestations of vitamin C deficiency 002314
Jeter sectcggrs 0 erase
Seorbutic rosary Rachitic rosary
i
| nmin nesses eens SE pars casemate
pushing in of sternum. SS ea Sp wok hss Eid atin Lo a vmeriioge ma SASS RS pps ein
Spinter hemorrhage vemearthvose.
Infante scurvy a/k/a Barlow's disease. Infants between b - 1 months are affected.
hen weaning starts, Yam GIRSBIROENGIES 50 they should be supplemented wih wiamn C.