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Bioenergetics & metabolism of carbohydrates, lipids, amino acids 1 Biologic oxidation 1.summary 2 O2 H 2O Carbohydrate, CO2 lipid, etc mitochondrion ATP enzymes 3 The concept of the biological oxidation 4 The three stages of biological oxidation 5 Nutriment materials Building block molecules Stage 1 Acetyl coenzyme A Stage 2 tricarboxylic acid cycle Stage 3 Oxidative phosphorylation 6 7 Nutriment materials biological oxidation H2O energy cell usage CO2 heat energy 8 Mitochondrion oxidation system 9 metabolism Carbohydrate, lipid, H2O, CO2 proteins FADH, NADH H2O, Mitochondrial oxidation system energy 10 the structure of mitochondrion Out membrane Inner membrane matrix 11 12 ATP synthase F0-F1 complex a large membrane –protein complex 13 14 15 16 respiratory chain 1. concept 2. constitutes four electron-carrier complexes 17 18 3. types NADH oxidation respiratory chain Succinate oxidation respiratory chain 19 4. The components & function of the each electron-carrier complexes in these two respiratory chain . A. complex Ⅰ ( NADH-Q reductase) 20 21 * Riboflavin proteins structure function types 22 23 24 * iron-sulfur proteins structure function 25 26 Fe 2 Fe 3 27 * ubiquinone,Q (coenzyme Q) structure function 28 29 30 B. complex Ⅱ ( succinic-Q reductase ) Fe-s AH2 A FAD FADH2 reduced Q Fe-S oxidized QH2 31 32 C. complex Ⅲ (Q-CytC reductase ) Contain: cytb562 cytb C cytc cytb566 C1 iron-sulfur proteins 33 * cytochromes, ( cyt ) a. structure 34 35 b. Classification: cytb562 cycb cytb566 cytc C C1 cyta a a3 36 c. the differences between each cytochromes the side chain linkage with apoenzyme Light –absorption spectra 37 d. function Single electron carriers iron in protoporphyrin prosthetic group 38 39 Fe 2 Fe 3 40 D. complexⅣ (CytC oxidase ) Contain: cyta,a3 CuA,CuB 41 42 5. The orders of these complexes in the respiratory chain A. A. NADH oxidation respiratory chain 43 • Compose of complex Ⅰ, Ⅲ, Ⅳ a. contain : + NAD ,FMN(Fe-S), CoQ,Cytb, c1,c , aa3,Cu,O2 44 b. order 45 46 47 B. Succinate oxidation respiratory chain • Compose of complex Ⅱ, Ⅲ, Ⅳ 48 a. contain : FAD+ (Fe-S), CoQ, Cytb, c1,c , aa3,Cu, O2 49 b. order 50 51 52 C. the evidences of the arrangement order • redox potential • light absorption spectra • inhibitors 53 54 55 • gentle treatment with detergents CoQ linked with complex Ⅰ ,Ⅱ, Ⅲ cytc linked with complex Ⅲ, Ⅳ 56 57 6. the process of the electron transportation 58 59 Oxidative phosphorylation A. concept 60 Metabolic oxidized materials products Respiratory chain 2H +1/2 O2 H 2O energy ADP H 3 PO4 ATP H 2 O 61 B. coupling sites of Oxidative phosphorylation 62 a. the methods for estimate of the coupling sites • the p/o ratio 63 64 • calculation formula of free energy change (standard reduction potential , E0’) ΔG0’= -nFΔE0’ 65 example: electrons pass from NADH(E0’=-0.32v ) to O2(E0’=+0.82v) ΔG0’= -296.6[0.82-(-0.32)]= -220 KJ NADH Q ΔG0’= -69.4 KJ CytB CytC ΔG0’= -34.7 KJ Cytaa3 O2 ΔG0’= - 102.3 KJ 66 67 C. The mechanism of Oxidative phosphorylation • The chemiosmotic hypothesis 68 69 D. factors effecting on oxidative phosphorylation • ADP/ATP • Thyroid hormone 70 E. inhibitors of oxidative phosphorylation • inhibitors of respiratory chain • uncoupler (2,4dinitrophenol) • inhibitors of oxidative phosphorylation (oligomycin) 71 72 73 74 The storage,release,& utilization of ATP A. the sources of high energy phosphate group in ATP 75 • oxidative phosphorylation • phosphorylation on substrate level 76 example: 1,3-diphosphoglycerate + ADP PEP + ADP phosphoglycerate + ATP EP + ATP PK succinylCoA + H3PO4 + GDP succinate+ CoA-SH + GTP + ADP ATP 77 B. the storage of ATP • phosphocreatin 78 79 C . the other nucleoside triphosphates 80 Electrons from cytosolic NADH enter mitochondrial by shuttles • glycerol phosphate shuttle • Malate-asparate shuttle 81 82 83