Metabolism - 10

Oxidative phosphorylation
1Glucose Oxidative Phosphorylation

Inner Mitochondrial Membrane

2NADH
4 complex 1 complex
2Pyruvate

Oxidation – e Remove
Coupling
2NADH Electron Transport chain ATP synthase
Hydride + H

2Acetyl CoA ½O2. + 2H ------- H2O ADP + Pi = ATP

6NADH Reduction

2FADH NADH
•
•
Proton motive forces
Mechanical force

Or Hydride = 2e + 1H
Oxaloacetate FADH
 Respirosome = Com-1,2,3
Electron transport chain & ATP synthase
Enzyme complex Respiratory enzyme Prosthetic group Inhibitor of ETC
Complex 1 (46 subunit) NADH dehydrogenase FMN, Fe-S Demerol, Rotenone ,
(NADH-Coenzyme Q Amobarbital ( Amytal)
reductase) Piericidin A, Barbutrirates
Complex 2 (4 subunit) Succinate dehydrogenase FAD , Fe-S Malonate
(Succinate - Coenzyme Q Carboxin
reductase)

Complex 3 (11 subunit) Cytochrome bc1 complex Heme , Fe- S Antimycin A

(Coenzyme Q-cytochrome British anti lewisite (BAL)
C reductase) Phenformin , Dimercaprol

Complex 4 (13 subunit) Cytochrome c oxidase Heme , Cu+ Cyanide , Azide

Carbon monoxide , H2S
Complex 5 ATP synthase F° - F¹ Oligomycin,Venturicidin
Aurovertin ,DCCD
Drop Mc is Anti-Bal phenomenon |
HACC VOAD ||
 Oxidation Phosphorylation
Complex – 1.
NADH + H  NAD+ Carrier Molecules
Hydride
FMN
CytC
Fe - S Complex -3 Complex 4 Complex -5
CO- Q  QH2 Fe – S --- Cyt C1 Cyt C
ADP + Pi  ATP
Fe – S
QH2 Cu a
Cyt B - L • 4 H  1ATP
FAD
QH • NADH = 10H = 2.5
Hydride Cyt a • FADH2 = 6 H = 1.5
Q Cyt B - H
FADH2  FAD+

Oxidation Q – cycle Cu b + Cyt a3 NADH

• Com-1 = 4
• Com-3 = 2
Succinate  Fumarate ½O2 + 2H -- H2O • Com – 4 = 4
Complex -2 FADH2
• Com -3 = 2
• Com-4 = 4
Q cycle complete in two phase
Inhibitor for Oxidation Phosphorylation
Q – Cycle
ATP synthase
 Shuttle System

Glycerol – 3 – Phosphate shuttle

Malate Aspartate shuttle
Glycerol 3 Phosphate shuttle
• In skeletal muscle and brain, the
reducing equivalent from
cytoplasmic NADH are transported
to Mitochondria as FADH2 through
Glycerol – 3 – phosphate shuttle.
• Hence only 1½ ATPs are generated
when this system is operating.
• In this shuttle, the enzyme called
Cytoplasmic Glycerol 3 Phosphate
Dehydrogenase 1 (GPDH – C)
converts dihydroxyacetone
phosphate to glycerol 3 Phosphate
by oxidizing one molecule of NADH
to NAD+.
 Malate Aspartate shuttle
• Mitochondrial Membrane is Impermeable to NADH.
• The NADH equivalent generated in Glycolysis are therefore to be
transported from cytoplasm to Mitochondria for oxidation.
• This is achieved by malate – aspartate shuttle or malate shuttle,
which operates mainly in liver , kidney and heart.
• The cycle is operated with the help of enzyme Malate
dehydrogenase ( MDH) and aspartate amino transferase.
• From one molecule of NADH in the mitochondria 2½ ATP
molecules are generated.
Uncoupling agent and Ionophore
Uncoupling
“ Uncoupling agents uncouple Oxidation from phosphorylation. They allow the
oxidation of NADH and FADH2 and reduction of O2 to continue at high levels but do
not permit ATP synthesis. Thus , electron transport continues unabated, but ATP
synthesis stops.”
• Most common uncoupling agents are....
1. 2,4-dinitrophenol (DNP)
2. Dicoumarol
3. Dinitrocresol
4. Carbonyl cyanide p (trifluorocarbonyl) phenylhydrazine ( FCCP)
5. Snake venoms phospholipase
6. Pentachlorophenol
7. Thermogenine
 Ionophores
• Ionophores are lipophilic molecules that bind specific cation and facilitate
their transport through the membrane.
• Ionophore uncouple electron transfer from oxidative Phosphorylation by
dissipating the electrochemical gradient across the mitochondrial
membrane.
• Valinomycine , an antibiotic, is an example of ionophores.
• Its addition makes inner mitochondrial membrane permeable for K+ . It
causes the movement of K + along the concentration gradient from
cytosol into the Matrix.
• It decreases the membrane potential component of pfm ( without a
direct effect on pH gradient) and thus ATP synthesis.
ATP – ADP exchange across the inner mitochondrial membrane

Inhibitors of ATP – ADP transporter

• Atracycloside
• Atracyloside is a plant toxin
• Inhibits adenine nucleotide
carrier
• Adenine nucleotide carrier
facillitates transport of ATP
and ADP
• Thus blocking ADP supply
prevents phosphorylation.