Proteins 1 ● Type, number, and sequence of

amino acids in a protein chain

- highly significant substances synthesized by ● Peptide bonds formed when
the liver from amino acids carboxyl group of one amino acid joins
- there are also proteins that are synthesized in the amino group of a second forming a
other locations molecule of water as by product
2 ● Twisted shape of the primary
Examples of proteins not produced by the liver: structure due to the rotation of bonds
 immunoglobulins – produced by plasma ● shape may be: alpha helix (spirals)
cells, not by the liver and beta-pleated (flat and corrugated)
 Adult hemoglobin – bone marrow 3 ● Three-dimensional structure that
 von Willebrand factor – platelets and forms when the amino acid chain folds
endothelial cells back to itself usually due to relatively
weak interactions of the interaction of
amino acids – building blocks of protein the side chains and is stabilized
through hydrophobic effect, ionic
- made up of continuous chain of carbon and attraction, hydrogen bonds and
nitrogen atoms joined together through peptide disulfide bonds.
bonds between adjacent amino acids ● The function and physical and
- Amphoteric molecules (react as acid or base) chemical properties of a protein
- effective antigens depends on its tertiary structure.
- provide 12 to 20% of total daily energy 4 ● Formed through the combination of
requirement of the body more than one polypeptide molecule
of protein subunits forming a
Function of proteins functional protein.
Catalyze biochemical Enzymes ● Not all proteins have a quaternary
reaction structure.
Provide structural, Collagen, keratin
cellular or body Protein Classification
support 1. Simple Proteins
Contraction of Actin, myosin Composed of amino acids linked by peptide
muscle bonds:
Neutralization of Immunoglobulins, a. Globular (transport protein)
foreign materials complement b. Fibrous (structural protein)
Transport of Albumin,
substances into the hemoglobin, 2. Conjugated Proteins
plasma transferrin It is composed of a protein and a prosthetic
Regulation of Hormones group:
metabolism a. Metalloproteins
Blood coagulation Coagulation factors EXAMPLE: Hemoglobin; Ceruloplasmin
Acid-base balance Albumin b. Lipoproteins
Maintenance of Albumin EXAMPLE: HLD; LDL; Chylomicrons
oncotic pressure c. Glycoproteins
d. Mucoproteins
The difference between glycoproteins and
mucoproteins is that glycoproteins contain

Protein Structure
 more proteins than carbohydrates while 3. Alpha 1 Antitrypsin
mucoprotein has more carbohydrates - main protein migrating in the alpha-1 region
e. Nucleoproteins and considered to be an acute phase reactant
EXAMPLE: Chromatin a.) Acute Phase Reactant - substance that
increases during an inflammation
Protein Catabolism and Nitrogen Balance In an inflammation, the white blood cell
● The biologic value of dietary protein is related produces elastase to eliminate bacteria; AAT
to the extent to which they provide all the neutralizes excess elastase; the elastase, if not
necessary amino acids. neutralized, will attack the cells of the lungs and
● The insufficient dietary quantities of even one the liver
amino acid can quickly limit the synthesis and ● lungs (Pulmonary Emphysema)
lower the body level of many essential proteins ● liver (Juvenile Cirrhosis)
● In normal healthy adults are in nitrogen
balance where intake and excretion are equal: 4. Alpha 1 Fetoprotein
a. Positive Nitrogen Balance - oncofetal antigen; the first proteins produced
The intake of nitrogen exceeds the loss thus net by the liver parenchyma (fetal) and detected in
protein synthesis exceeds; the anabolism is the amniotic fluid
higher than catabolism a.) acts as a marker for possible defects in the
EXAMPLE: Wound Healing; Pregnancy baby
b. Negative Nitrogen Balance ↑ AFP – neural tube defect
There is more nitrogen excreted than ↓ AFP – Down syndrome
incorporated in the body; the catabolism is b.) acts as a marker for tumors
higher than anabolism. ↑ AFP in adults - hepatic cancer or testicular
carcinoma
Significant Plasma Proteins
1. Prealbumin or Transthyretin 5. Acid Glycoprotein
- migrates ahead of albumin - It is the transporter protein for progesterone
a.) To transport of thyroxine or T4 and vitamin A and also called orosomucoid
or retinol
b.) As a marker of malnutrition 6. Alpha 2 Macroglobulin
It has a very short half-life which is 2 days; in It is the largest non-immunoglobulin protein
malnutrition, the prealbumin is heavily affected. a. Nephrotic Syndrome
In comparing serum or plasma to cerebrospinal - The protein is being retain due to high
fluid, the prealbumin is higher in CSF molecule weight
- It is produced ten-fold thus higher levels
2. Albumin - The production of this protein increases to
- most abundant protein in the blood, compensate the loss of other protein to
comprising 50% or more of total protein maintain oncotic pressure
a.) acts as a transport protein for unconjugated
bilirubin or B1, calcium ion and magnesium ion 7. Ceruloplasmin
b.) major contributor to oncotic pressure - transport protein of copper (6); it is a protein
When protein is decreased, the oncotic with enzymatic activity or ferroxidase activity
pressure drops and water is leaked from the a.) converts stored iron to transport iron
vessels causing edema due to effusion or water Wilson’s Disease - ceruloplasmin deficiency
accumulation
 WILSON’S DISEASE MENKE DISEASE
Ceruloplasmin Severe Copper 11. Hemopexin
Deficiency Deficiency - To bind heme
Low Serum Low Serum ● It is associated with intravascular and
Ceruloplasmin Ceruloplasmin extravascular hemolysis (decreases in both
Low Serum Copper Low Serum Copper hemolysis)
Highly Elevated Urine Low or High Urine
Copper Copper INTRAVASCULAR HEMOLYSIS
With Copper The unbound(free) hemoglobin is broken down
Deposits in Brain, No Copper Deposits into heme and globin, the heme is then bound
Eyes, Liver to hemopexin which leads to its decrease
Kayser Fleischer Ring
or Copper Deposit in Kinky Hair EXTRAVASCULAR HEMOLYSIS
the Eye The red blood cells are destroyed in the liver or
spleen which releases hemoglobin and
8. Haptoglobin degraded into heme and globin; the heme is
- It binds free hemoglobin; ↓ intravascular then bound to hemopexin. No free hemoglobin
hemolysis is present.
● In the destruction of red blood cells, the
hemoglobin is released on the blood vessels; INTRAVASCULAR EXTRAVASCULAR
the haptoglobin binds to free hemoglobin. HEMOLYSIS HEMOLYSIS
● In severe cases, as haptoglobin is depleted, ↓ Haptoglobin Normal Haptoglobin
nothing will hold hemoglobin down so it is ↓ Hemopexin ↓ Hemopexin
excreted through urine (hemoglobinuria) With Hemoglobinuria No Hemoglobinuria
Intravascular hemolysis is characterized by
hemoglobinuria 12. Complement Proteins
● They contribute to the immune system with
9. Transferrin an end product of cellular lysis
- It is considered the major protein migrating in ● C3 (Most Abundant)
the beta region and the transport protein for
iron (2) 13. C-Reactive Proteins
● aka siderophilin; it is a negative acute phase ● They are found to be reacting with the C-
reactant which decreases during inflammation substance found on the cell wall of
pneumococci
Negative Acute Phase Reactants ● sensitive but nonspecific acute phase
a) Albumin 10. Fibr reactants increasing 1000x in inflammation
b) Prealbumin ino ● The levels will be higher in bacterial infections
c) Transferrin gen compared to other types of infections
● It is It is associated with the increased risk of
the cardiovascular disease
most abundant coagulation factor in the blood
and also called Factor I 14. Immunoglobulin or Antibody
● It is an acute phase reactant and one of the ● It was previously called gammaglobulins
most affected substances in disseminated because it was migrating in the gamma region
intravascular coagulation (consumption of ● It is one of the proteins not produced by the
fibrinogen) liver. plasma cells synthesize them during the
secondary or late response
IgM - It is a pentamere thus the biggest and an Monoclonal Gammopathy
acute phase antibody and is firstly produced in Total Protein: Absolute Increased
infection Albumin: Normal / Decreased
IgG - It is a monomer and the most abundant Globulin: Increased
but is anamnestic which is the second antibody A/G Ratio: Decreased
produced in infection  This condition is characterized by increased
IgA in the production of gammaglobulin
IgE
IgD HYPOPROTEINEMIA
Acute and Chronic Inflammation
TOTAL PROTEIN, ALBUMIN, GLOBULIN, AG Total Protein: Decreased
RATIO Albumin: Decreased
 It entails total protein and albumin Globulin: Increased
values to calculate for globulin levels A/G Ratio: Decreased
and A/G ratio
Hepatic Cirrhosis
Globulin=Total Protein− Albumin Total Protein: Decreased
Albumin Albumin: Decreased
AG Ratio=
Globulin Globulin: Increased
A/G Ratio: Decreased
a. AG Ratio  It results in the liver’s inability to produce
normally 1 to 1.8; the albumin and protein; the IgM has increased production
globulin should approximately the
same Nephrotic Syndrome
a) Decreased AG Ratio Total Protein: Decreased
I. Decreased Albumin Albumin: Decreased
II. Increased Globulin Globulin: Normal
EXAMPLE: Kidney Diseases; Liver Diseases A/G Ratio: Decreased
b) Increased AG Ratio  The alpha 2 macroglobulin is not removed
I. Increased Albumin from the body but is overproduced thus
There is no condition that the globulin levels does not decrease
albumin is only increased
II. Decreased Globulin Immunodeficiency
 The albumin is directly proportional to the Total Protein: Decreased
AG ratio while indirectly proportional to Albumin: Normal
globulin Globulin: Decreased
A/G Ratio: Increased
HYPERPROTEINEMIA  Deficient in immunoglobulins
Dehydration
Total Protein: Relative Increased
Albumin: Relative Increased A. PROTEIN ELECTROPHORESIS
Globulin: Relative Increased BANDS PROTEINS
A/G Ratio: Normal Albumin
Albumin
 The total protein is only increased due to (62%)
the loss of water causing a relative increase α-1 (4) Alpha 1 Antitrypsin (Main
Protein)
 Alpha 1 Fetoprotein
Acid Glycoprotein
Alpha 2 Macroglobulin 3. Hypogammaglobulinemia
(Main Protein) a. Highly Decreased Gamma
α-2 (8.6%)  There is decrease or failure in the
Ceruloplasmin
Haptoglobin production of immunoglobulins which can
Transferrin (Main Protein) be seen in immunodeficient states
Hemopexin EXAMPLE: Bruton’s Disease;
β (12.5%) Complement Proteins Agammaglobulinemia; HIV or AIDS
Fibrinogen
C-Reactive Proteins 4. Hepatic Cirrhosis or Polyclonal
Gammopathy
Immunoglobulins
γ (12.9%) a. Decreased Albumin
C-Reactive Proteins
b. Highly Increased Gamma
 The albumin is decreased due to decrease
 In a serum specimen, the fibrinogen in the
or failure of production by the liver
beta band will no longer be found
 The increased production “fast migrating”
 The most anodal protein is albumin
immunoglobulins or IgM creates a bridging
because it is the protein that is first to
effect towards the beta region thus called
migrate to the anode
beta-gamma bridging.
 The prealbumin actually arrives first than
albumin but it is not considered to be a
5. Monoclonal Gammopathy
main protein
a. Decreased Albumin
b. Marked Increase Gamma with
1. Immediate Response Pattern
Narrow Band
a. Decreased Albumin
 There is a gamma spike wherein the
b. Increased Alpha 1
gamma region is a narrow spike
c. Increased Alpha 2
EXAMPLE: Multiple Myeloma or Increased
d. Increase in Beta region
Immunoglobulin Light Chain; Waldenstrom
 It is also known as acute inflammation; the
Macroglobulinema or Increased IgM
decreased albumin is because it is a
negative acute phase reactant
MULTIPLE WALDENSTROM
 The increase alpha 1 and 2 and beta
MYELOMA MACROGLOBULINEMA
regions is because acute phase reactants
Increased Increased IgM
are usually migrating in these regions
Immunoglobulin
Light Chains due to
2. Delayed Response Pattern
Plasma Cell
a. Decreased Albumin
Neoplasm
b. Increased Alpha 1
c. Increased Alpha 2 Common Bence Less Common Bence
d. Increase in Beta region Jones Proteinuria Jones Proteinuria
e. Increased Gamma Bone Involvement No Bone Involvement
 It is also known as chronic inflammation; Hypercalcemia No Hypercalcemia
the gamma region is increased due to the
production of immunoglobulins 6. Nephrotic Syndrome
a. Highly Decreased Albumin
b. Highly Increased Alpha 2
c. Highly Decreased Gamma
  The marked decrease in albumin and
other globulins is due to urinary loss; 2. Kjeldahl Method
the increased alpha 2 region is due to  It is a reference method involved in the
retention of alpha 2 macroglobulin digestion of protein and measurement of
producing in 10 folds nitrogen content
 There may be an alpha 2 spike or  It assumes that the average nitrogen
increase in alpha 2 and beta region; the content of 16%
beta region may be increased due to
the retention of hemopexin 3. Direct Optical Method or UV Method
 It is based on the absorbance of proteins at
7. Protein Losing Enteropathy the UV spectrum
a. Highly Decreased Albumin a. 210nm: Peptide Bonds
b. Increased Alpha 2 b. 280nm: Amino Acids – Tryptophan
c. Decreased Gamma
 It is characterized by losing proteins in 4. Dye Binding Techniques
the intestines or is not absorbed by the  It is mainly used for albumin
proteins determination; it binds to the dye
causing a shift in the maximum
8. Alpha 1 Antitrypsin Deficiency absorption
a. Decreased Alpha 1 a. Bromcresol Green (BCG)
 The deficiency in alpha 1 antitrypsin is  It is mostly commonly used which is
due to pulmonary emphysema or highly sensitive but overestimates low
juvenile cirrhosis; the pattern is called albumin levels
alpha 1 flat curve b. Bromcresol Purple (BCP)
 It is the most sensitive, specific and
ERRORS IN PROTEIN ELECTROPHORESIS precise
1. Inadvertent Use of Plasma Instead of c. HABA (2,4’-Hydroxyazobenzene
Serum Benzoic Acid)
 The narrow distinct band in the beta  It is affected by a lot of interference
region due to the presence of such as salicylates and bilirubin
fibrinogen d. Coomassie Brilliant Blue (CBB)
2. Hemolysis  It is used to determine proteins in
 There is a blip in the pattern in the late other body fluids specifically for
alpha 2 or early beta region due to the cerebrospinal fluid electrophoresis
presence of free hemoglobin  There is limited linearity with unequal
 There is a small blip in the alpha 2 sensitivity for individual proteins
region due to hemoglobin-haptoglobin
complexes [Link] Precipitation or Turbidimetry
Techniques
B. PROTEIN DETERMINATION METHODS  The proteins are precipitated as fine
1. Biuret Method particles; the turbidity is measure
 It is a routine method based on the spectrophotometerically
reaction of peptide bonds and cuprous ions  The amount of protein formed is directly
forming a violet complex measured at proportional to the amount of precipitate
540nm formed
 It requires at least two peptide bonds and  It is rapid and easy to use but unequal
an alkaline medium sensitivity for individual protein
a. Sulfosalicylic Acid
 It is more sensitive for albumin and
sodium sulfate is added to better
precipitate globulins
b. Tricholoracetic Acid
 It is the preferred acid for
cerebrospinal fluid; it detects both
albumin and globulin