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Matrix
H+ + NADH NAD+ + 2H+
F1 Fo
2 e Q
III IV
++
4H+ 4H+
cyt c
2H+
3H+
Intermembrane Space
The Chemiosmotic Theory of oxidative phosphorylation, for which Peter Mitchell received the Nobel prize:
Coupling of ATP synthesis to respiration is indirect, via a H+ electrochemical gradient.
Protons (H+) accumulate intermembrane space creating an electrochemical potential difference, proton gradient or electrochemical gradient. This proton motive force (PMF) drives the synthesis of ATP by ATP synthase complex.
IMM- Inner mitochondrial membrane IMS- Inter membrane space OMM- outer mitochondrial membrane
CHEMIOSMOTIC THEORY
Peter mitchel
H+ H+ H+ 4H+
I
H+ +
4H+
III
H+
2H+
Iv
H + H+ H+
2e-
4H+
H+ H+ ADP+Pi H+ H+ 4H+ H+ + 4H H+ + V H+ H + H+ H + H+ + ATP H H+ H ++ + H H+ H H H+ + H+ MATRIX H + H + H+ H+ + H + H H+ H+ H + IMM H+ H H+ H + H + H+ H+ + Complex I, III and IV H IMS are proton pumps H+ OMM
4H+
H+ + H H+ H+ H+
Uses proton gradient to make ATP Protons pumped through channel on enzyme
From intermembrane space into matrix ~4 H+ / ATP
NADH
10 H+ X 1 ATP = 2.5 (3) ATP 4 H+ FADH2 6 H+ X 1 ATP = 1.5 (2) ATP 4 H+
Boyer s binding change mechanism: ATP synthase is a protein assembly in the inner
mitochondrial membrane.
ADP + Pi F1 Fo
ATP
4H 3
+
matrix
intermembrane space H+ H+ H+ H+ H + H + H + H +
subunits occur in 3 forms O form (Open form). It has low affinity for substrates ADP +Pi
L form (loose form). Can bind substrates ADP and Pi but catalytically it is inactive. T form (Tight form). Binds substrates ADP + Pi tightly and catalyses ATP synthesis.
When protons pass through the disk of C subunits of F0 unit it causes rotation of sub unit. The subunits which are fixed to the membrane donot rotate. ADP & Pi are taken up sequentially by the subunits which undergo conformational changes and form ATP.
ETC - inhibitors
Complex I :
Rotenone, Peircidin, Amytal, Barbiturates ComplexII: Carboxin,Thenoyltrifluroacetone,malonate Complex III : site II of ATP synthesis inhibitors
Antimycin, Myxothiazol , stigmatellin Complex IV: site III of ATP synthesis inhibitors
Uncouplers will allow oxidation to proceed but energy instead of being trapped as ATP is dissipated as heat.
These protonated uncouplers due to their lipophilic nature rapidly diffuse across the membrane into
2-4 dinitrophenol a classical uncoupler electrons from NADH to oxygen proceeds normally but ATP not formed as proton motive force across inner mitochondrial membrane is dissipated .
2. Penta chloro phenol 3. Dinitro cresol 4.Bilirubin 5.Thyroxine-Physiological uncoupler 6.Valinomycin 7.Nigericin
Note: They are Lipophilic
matrix
Intermembrane space H+ H+
H+ H+
H+ H+
H+ H+ H+ H+
Physiological Uncouplers 1.Excessive thyroid hormones 2. Unconjugated hyper bilirubinaemia 3. In high doses aspirin uncouple oxidative phospharylation which explains fever that accompanies toxic over dosage of these drugs.
Uncoupling proteins
UCPs occur in the inner mitochondrial membrane of mammals, including humans. UCPs create a proton leak, that is they allow protons to re-enter the mitochondrial matrix without
UCP1, also called thermogenin, is responsible for the activation of fatty acid oxidation and heat production in the brown adipocytes of mammals. Brown fat , unlike the more abundant white fat, uses almost 90% of its respiratory energy for thermogenesis in response to cold, at birth,etc.
Oligomycin acts through one of the proteins present in F0 - F1 stalk . Oligomycin blocks the synthesis of ATP by preventing the movement of protons through ATP synthase.
The regulation of the rate of oxidative phosphorylation by ADP level is called respiratory control. The ADP level increases when ATP is consumed and so oxidation is coupled to the utilization of ATP. Under physiological conditions, electron transport is tightly coupled to oxidative phosphorylation.
Electrons do not usually flow through the electron transport chain to O2 unless ADP is simultaneously phosphorylated to ATP.
In the presence of excess substrate and Oxygen, respiration continues until all ADP is converted to ATP. After that the respiration rate or utilization of oxygen decreases In the presence of adequate oxygen and substrate, ADP becomes rate limiting; it exerts a control over the entire oxidative phosphorylation process
Oxidation cannot proceed via ETC without simultaneous phosphorylation of ADP. Chance & Williams defined 5 conditions that can control rate of respiration.
Generally most cells in the resting state are in state 4 , and respiration is controlled by the availability of ADP. The availability of inorganic phosphate could also influence the respiration. As respiration increases (Exercise) cell approaches state 3 ( ETC working to its full capacity ) or state 5 ( Availability of O2 is a limiting factor ). ADP / ATP transporter may also be a rate limiting factor