Electron transport chain - NOTES
Def: The transfer of electrons from reduced co-enzymes thro’ the respiratory chain to oxygen
is known as Biological oxidation. Energy is released during this process, which is trapped as
ATP.
This coupling of oxidation with phosphorylation is called ‘Oxidative Phosphorylation’.
Organization of ETC:
- In ETC, the electrons(e-) are transferred from NADH to a chain of electron carriers.
The e- flow from electronegative components to more electropositive components.
- Components of ETC are located in the inner membrane of Mitochondria.
Complexes of ETC:
Complex I: NADH Dehydrogenase – contains the prosthetic group FMN, Fe-S cntrs
Complex II: Succinate Q Reductase – contains FAD, cyt b560, 3 Fe-S cntrs
Complex III: Cytochrome Reductase – contains cyt bH, cytbL, cyt c1, Fe-SRieske
Complex IV: Cytochrome Oxidase – contains cyt a, cyt a3, CuA, CuB
Mobile carrier: 1. Co-enzyme Q 2. Cytochrome c
Shuttle pathways:
a) Malate Aspartate shuttle:
- Mitochondrial membrane is impermeable to NADH.
- Present in liver, kidney, heart.
- NADH equivalents generated are transported from cytoplasm to mitochondria.
- One molecule of NADH generates 2.5 ATP.
b) Glycerol-3-phosphate shuttle: ( PN pge: 154; fig: 8.14 )
- Present in skeletal muscle & brain
- Reducing equivalents from cytoplasmic NADH are transported to mitochondria as
FADH2.
- One molecule of FADH2 generates 1.5 ATP.
Complex I: NADH Dehydrogenase :
- Components: a) FMN b) Iron-sulphur protein
- NADH is the donor of electrons.
- NAD prosthetic group are: Glyceraldehyde 3-P DH, Isocitrate DH, Malate DH
- Flow of electrons: NADH FMN Fe-S CoQ
- Free energy of -12 kCal/ mol is released, which is utilised to drive 4 protons out of the
mitochondria.
- It is the site 1 for ATP formation
Complex II: Succinate Q Dehydrogenase
- Components: a) FAD b) Iron-sulphur protein
- FADH2 is the donor of electrons.
- FAD prosthetic group are: Succinate DH, Fatty acyl CoA DH & Glycerol P DH
- Flow of electrons: FADH2 FAD Fe-S CoQ
- This step does not liberate enough energy to act as proton pump..
- No ATP formation.
Co-enzyme Q: is a quinone derivative having a long isoprenoid tail.
It accepts a pair of electron, Ubiquinone is reduced to semiquinone and then to Quinol.
Q cycle facilitates the switching from 2 electron carrier to single electron carrier Cytochrome c.
It occurs in 2 steps.
Step (1) one electron from QHA is donated to Cyt c1 to cytc c via Reieske Fe-S and 2nd electron to Q to
form semiquinone via cyt bL & cyt bH with 2 H+ being released into intermembrane space.
Step (2) occurs with 2nd QH2, the 2nd electron is donated to semiquinone, reducing it to QH2 by taking
2H+ from the matrix
Complex III: Cytochrome Reductase
- Components: a) Iron-sulphur protein b) cytochrome b
c) Cytochrome c1
- Flow of electrons: CoQ FeS Cyt b Cyt c1 Cytochrome c
- Free energy of -10 kCal/ mol is released, which is utilised to drive 4 protons out of the
mitochondria.
- It is the site 2 for ATP formation.
Cytochrome c
It is the peripheral membrane protein containing one heme prosthetic group.
Cytochrome c collects electrons from complex III and delivers to complex IV.
Complex IV: Cytochrome Oxidase
- Components: a)cytochrome a b) cytochrome a3 c) Cu A d) Cu B
- Flow of electrons: Cytochrome c cyt a-a3 O2
- Free energy of -24 kCal/ mol is released, which is utilised to drive 2 protons out of the
mitochondria.
- It is the site 3 for ATP formation.
Energetics of ATP synthesis:
Energy of electron transfer is used to drive protons out of the matrix by the complexes I, III & IV.
3 protons are required for one ATP formation. When one NADH transfers its electrons to oxygen to
oxygen, 10 protons are pumped out and 2.5 ATP can be synthesized.
Similarly, electron transferred from FADH2 to oxygen, 6 protons are pumped out and 1.5 ATP can
be synthesized.
Chemiosmotic theory
Def: The coupling of oxidation with phosphorylation is termed as Oxidative phosphorylation.
Mechanism:
- The proton pumps (complex I, III, IV) expels H+ from inside to outside of the inner
mitochondrial membrane.
- Protons accumulate outside the membrane, so there is high H+ concentration.
- Resulting in generation of a proton gradient across the membrane, creating an
electrochemical potential difference.
- pH outside the mitochondrial inner membrane is 1.4 units lower than inside.
- This proton motive force drives the synthesis of ATP by ATP synthase complex.
- ATP synthase or complex V: is a multisubunit transmembrane protein. fig: 8.9
- F0 unit – spans the inner mitochondrial membrane.
- F1 unit – projects into the matrix. It catalyses ATP synthesis.
- Protons entering the system causes conformational changes and induces ATP
synthesis.
Inhibitors of ETC
A) site specific inhibitors:
1) site : Complex I to CoQ inhibitors
1. Alkylguanides, hypotensive drugs
2. rotenone, insecticides, fish poison
3. Chlorpromazine, tranquiliser
4. barbiturates, sedatives
5. piericidin, antibiotic
2) site between Complex II and CoQ
1. Carboxin
2. Malonate, competitive inhibitor of Succinate DH
3) site : Complex III to Cytochrome c
1. British anti lewisite (BAL), antidote of war gas
2. napthoquinone
4) site : Complex IV Inhibitors
1. carbonmonoxide, inhibits cellular respiration
2. cyanide
3. azide
4. Hydrogen sulphide
B) Inhibitors of Oxidative Phosphorylation
1. Atractyloside, inhibits Translocase
2. Oligomycin, inhibits flow of protons thro’ Fo
3. Ionophores eg. Valinomycin
Increases the permeability of lipid bilayer & dissipate the proton gradient.
C) Uncouplers of Oxidative phosphorylation
Uncouplers are chemical substances that allow electron transport chain in mitochondria.
But prevent the phosphorylation of ADP to ATP by uncoupling the essential linkage between
electron transport and phosphorylation for ATP synthesis.
Uncoupling agents are lipid soluble that diffuses thro’ mito membrane & binds H+ ion.
Membrane is abnormally permeable to protons.
Prevents the formation of proton gradient which is required for ATP formation.
Allows oxidation to proceed but the energy is dissipated as heat.
Eg:
1. 2,4 Dinitrophenol (DNP)
2. Dicuomarol
3. Chlorocarbonylcyanide phenylhydrazone
4. Salicylate
5. Physiological uncouplers : bilirubin, FFA, Thyroxine .
6. Natural uncoupler: Thermogenin present in adipocytes.
Substrate Level Phosphorylation :
DEF:-Substrate-level phosphorylation is the production of ATP from ADP by a direct transfer of a
high-energy phosphate group from a phosphorylated intermediate metabolic compound.
Does not involve the Electron Transport chain in the mitochondria.
Examples:
Glycolysis
TCA cycle
