Electron transport chain,oxidative phosphorylation & mitochondrial transport systems. M.Kohutiar, B.Sopko
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1 Electron transport chain,oxidative phosphorylation & mitochondrial transport systems. M.Kohutiar, B.Sopko
2 Content 1. Structure of mitochondria Mitochondrial transport systems 2. Electron transport 3. Parts of Electron transport chain 4. Proton gradient and synthesis of ATP 5. Uncoupling
3 Metabolism ANABOLIC CATABOLIC ΔG > 0 ΔG < 0
4 Gain of electron transport chain C H O 6O 6CO 6H O G 2823kJ / mol
5 Gain of electron transport chain 1 NADH H O NAD H O n(adp P ) i n(atp)
6 Gain of electron transport chain C H O 6O 6CO 6H O G 2823kJ / mol C H O 6O 6CO H 24e 6O 24H 24e 12H O 2 2
7 Gain of electron transport chain -0,4 NADH COMPLEX I FADH 2 COMPLEX II CoQ COMPLEX III cytochrome c COMPLEX IV 0,8
8 Gain of electron transport chain -0,4 NADH COMPLEX I FADH 2 COMPLEX II CoQ COMPLEX III cytochrome c COMPLEX IV 0,8
9 Gain of electron transport chain -0,4 NADH COMPLEX I FADH 2 COMPLEX II CoQ COMPLEX III cytochrome c COMPLEX IV 0,8 ΔE > 0
10 Gain of electron transport chain 1 NADH H O NAD H O n(adp P ) i n(atp) -0,4 NADH COMPLEX I FADH 2 COMPLEX II CoQ COMPLEX III cytochrome c COMPLEX IV 0,8
11 GLYCOLYSIS glucose glucose-6-phosphate glyceraldehyde-3-phosphate NAD + NADH 1,3-bisphosphoglycerate pyruvate NAD + acetyl-coa NADH
12 GLYCOLYSIS glucose glucose-6-phosphate glyceraldehyde-3-phosphate NAD + acetyl-coa NADH 1,3-bisphosphoglycerate NADH citrate ocalacetate pyruvate NAD + NAD + isocitrate NAD + acetyl-coa NADH malate fumarate CITRATE CYCLE 2-oxoglutarate NADH FADH 2 sukcinate sukcinyl-coa NAD + FAD NADH
13 Mitochondrial anatomy
14
15 ATP ADP translocase
16 Cofactors structure
17
18
19 Glycerolphosphate shuttle cytosol mitochondria CH 2 OH HC OH CH 2 O P NAD + glycerol-3-phosphate NADH CH 2 OH C O CH 2 O P dihydroxyacetone phosphate
20 Glycerolphosphate shuttle cytosol mitochondria CH 2 OH HC OH CH 2 O P glycerol-3-phosphate NAD + NADH CH 2 OH C O CH 2 O P dihydroxyacetone phosphate CH 2 OH HC OH CH 2 O P glycerol-3-phosphate FAD FADH 2 CH 2 OH C O CH 2 O P dihydroxyaceton phosphate
21 Malate-aspartate shuttle cytosol mitochondria HO NAD + NADH O HC CH 2 COOH COOH MDH C COOH CH 2 COOH malate oxalacetate
22 Malate-aspartate shuttle cytosol mitochondria HO NAD + NADH O HC CH 2 COOH COOH MDH C COOH CH 2 COOH malate oxalacetate malate oxalacetate HO HC COOH CH 2 COOH NAD + MDH NADH O C COOH CH 2 COOH
23 Malate-aspartate shuttle cytosol mitochondria HO NAD + NADH O HC CH 2 COOH COOH MDH C COOH CH 2 COOH malate oxalacetate malate oxalacetate HO HC COOH CH 2 COOH NAD + MDH NADH O C COOH CH 2 COOH AST Glu Asp ketoglutarate O C COOH (CH 2 ) 2 COOH
24 Malate-aspartate shuttle HO NAD + NADH O O HC AST CH 2 COOH COOH MDH C COOH CH 2 COOH Glu Asp C COOH (CH 2 ) 2 COOH cytosol malate oxalacetate a-ketoglutarate mitochondria malate oxalacetate a-ketoglutarate HO HC COOH CH 2 COOH NAD + MDH NADH O AST O C COOH CH 2 COOH Glu Asp C COOH (CH 2 ) 2 COOH
25 Electron transport NAD H 2e NADH E 0 0,32V 1 0 O2 2H 2e H2O E 2 0,82V
26 Electron transport NAD H 2e NADH E 0 0,32V 1 0 O2 2H 2e H2O E 0,82V 2 E 0 0,82-(-0,32) 1,14 V ΔG 0 zfδf 0 2 F1, kj/mol
27 Electron transport ADP P ATP ΔG 0 30,5 kj/mol
28 Electron transport ADP P ATP ΔG 0 30,5 kj/mol 330, %
29 Electron transport ADP P ATP ΔG 0 30,5 kj/mol 330, % ΔE 0 0 ΔG zf F 0,16 V
30 Electron transport NAD CoQ ox NADH CoQ red ΔE 0 0,36 V ΔG 0-69,5 kj/mol
31 Electron transport NADH CoQ ox NAD CoQ red ΔE ΔG 0 0 0,36 V -69,5 kj/mol CoQ red cytc ox CoQ ox cytc red ΔE 0 0,19 V ΔG 0-36,7 kj/mol
32 Electron transport NADH CoQ ox NAD CoQ red ΔE ΔG 0 0 0,36 V -69,5 kj/mol CoQ red cytc ox CoQ ox cytc red ΔE 0 0,19 V ΔG 0-36,7 kj/mol cytc red 1 2 O 2 cytc ox H 2 O ΔE ΔG 0 0 0,58 V -112 kj/mol
33 Electron transport FADH 2 CoQ ox FAD CoQ red ΔE 0 0,015 V ΔG 0-2,9 kj/mol
34 -0,4 NADH COMPLEX I FADH 2 COMPLEX II CoQ COMPLEX III cytochrome c COMPLEX IV 0,8
35 -0,4 FADH 2 COMPLEX II NADH COMPLEX I CoQ COMPLEX III cytochrome c ADP ATP ADP ATP 0,8 COMPLEX IV ADP ATP
36 -0,4 NADH COMPLEX I ADP rotenone ATP FADH 2 COMPLEX II CoQ COMPLEX III ADP antimycin A ATP cytochrome c 0,8 COMPLEX IV ADP ATP cyanide
37 [O 2 ] čas Time
38 [O 2 ] NAD Time
39 [O 2 ] NAD + 1 rotenon čas
40 [O 2 ] NAD + 1 rotenone 2 3 FADH time
41 [O 2 ] NAD + 1 rotenone 2 3 FADH 2 antimycin A Time
42 [O 2 ] NAD + 1 rotenone 2 3 FADH 2 antimycin A 4 5 cyanide 6 Time
43 Components of electron transport chain Complex I: NADH ubiquinonreductase NADH CoQ ox NAD CoQ red ΔE ΔG 0 0 0,36 V -69,5 kj/mol
44 FeS centres
45 Components of electron transport chain Complex II: succinate ubiquinon reductase FADH 2 CoQ ox FAD CoQ red ΔE 0 0,015 V ΔG 0-2,9 kj/mol
46 Components of electron transport chain Complex III:ubiquinol-cytC-reductase CoQ red cytc ox CoQ ox cytc red ΔE 0 0,19 V ΔG 0-36,7 kj/mol
47 Cytochromes
48 Components of electron transport chain Complex IV: cytochrome c oxidase cyt c(fe ) 4H O2 4 cyt c(fe ) 2H2O
49 Oxidative phosphorylation chemiosmotic theory (Mitchell) OXPHOS requires intact mitochondrial membrane Inner membrane is impermeable for some ions Electron transport is accompanied by transport of H +, and development of measurable gradient Compounds which increase the membrane permeability do not affect the electron chain but inhibit ATP synthesis
50 high H + H + H + H + H low H + ADP ATP
51 Proton gradient ΔG RT[pH in ph ex ] zfδ ΔG 0 21,5 kj/mol
52 Proton gradient ΔG RT[pH in ph ex ] zfδ ΔG 0 21,5 kj/mol Cytosolic side
53 Mechanism of redox loop Interimemb. space inner membrane e - matrix QH 2 H + QH.
54 Mechanism of redox loop Interimemb. space inner membrane e - matrix QH 2 QH 2 H + QH.
55 Mechanism of redox loop Interimemb. space inner membrane matrix e - H + QH 2 QH 2 H + e - cyt b k QH. QH.
56 Mechanism of redox loop Mezimemb. prostor inner membrane matrix e - H + QH 2 QH 2 H + e - cyt b k QH. QH. H + e - cyt c Q
57 Mechanism of redox loop Interimemb. space inner membrane matrix e - H + QH 2 QH 2 H + e - cyt b k QH. QH. H + e - Q Q cyt c
58 Mechanism of redox loop Interimemb. space inner membrane matrix e - H + QH 2 QH 2 H + e - cyt b k QH. QH. e - cyt b T H + e - Q Q cyt c
59 Mechanism of redox loop Interimemb. space inner membrane matrix e - H + QH 2 QH 2 H + e - QH. cyt b k e - cyt b T e - QH. H + e - Q Q H + cyt c
60 Structure of Complex V
61 3D Structure of Complex V
62 Schematic Structure of Complex V
63 Dam
64 Dam
65 Protonmotiv force and ATP synthesis
66 ATP synthesis
67 Regulation of oxidative phosphorylation 1 3 ADP P NAD cyt c 2 i NADH cyt c ATP
68 Regulation of oxidative phosphorylation ATP c cyt NAD 2 1 P ADP c cyt NADH i 3 ADP ADP.P cyt c cyt c NADH NAD K i
69 Regulation of oxidative phosphorylation ATP c cyt NAD 2 1 P ADP c cyt NADH i 3 K ATP ADP.P NAD NADH cyt c c cyt i i ADP.P ATP cyt c cyt c NADH NAD K
70 Uncoupling
71 Aerobic vs. Anaerobic ATP production Oxidation of 2 e - NADH. 2,5 mol ATP with consumption of 0,5 mol O 2 1 mol of the substrate is oxidised via Complexes I, III, and IV Oxidation of 2 e - FADH 2 1,5 mol ATP 1 mol of the substrate is oxidised via Complexes II, III a IV Total oxidation of 1 mol of glucose yields mol ATP,
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