Krebs Cycle, Electron Transport and Oxidative Phosphorylation

Yulia Suciati

SIKLUS KREBS

SIKLUS ASAM SITRAT

Terjadi didalam matriks mitokondria Proses ini bersifat aerobik Fungsi utama siklus asam sitrat (siklus krebs) a/ bekerja sbg lintasan akhir bersama untuk oksidasi KH, Lipid, Protein. Glukosa, as. Lemak, AA, dimetab. Mjd asetil KoA atau senyawa antara di SAS.

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Step 1: Condensation In step 1 of the Krebs cycle, the two-carbon compound, acetyl-S-CoA, participates in a condensation reaction with the four-carbon compound, oxaloacetate, to produce citrate:

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Step 2. Isomerization of Citrate step 2 involves moving the hydroxyl group in the citrate molecule so that we can later form an a-keto acid

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Step 3: Generation of CO2 by an NAD+ linked enzyme The Krebs cycle contains two oxidative decarboxylation steps; this is the first one The reaction is catalyzed by the enzyme Isocitrate dehydrogenase

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Step 4: A Second Oxidative Decarboxylation Step This step is performed by a multi-enzyme complex, the a-Ketoglutarate Dehydrogenation Complex

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Step 5: Substrate-Level Phosphorylation

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Step 6: Flavin-Dependent Dehydrogenation

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Step 7: Hydration of a Carbon-Carbon Double Bond

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Step 8: A Dehydrogenation Reaction that will Regenerate Oxaloacetate

HASIL AKHIR S.A.S

12 molekul ATP terbentuk pada setiap kali putaran S.A.S Sejumlah ekuivalen pereduksi akan dialihkan kpd rantai pernafasan dlm membran dalam mitokondria.

VITAMIN YG PENTING PD S.A.S

Riboflavin, dlm bentuk FAD (Flavin Adenin Dinukleotida) Niasin, dlm bentuk NAD (Nikotinamide Dinukleotida) Tiamin, dlm bentuk TPP (Tiamin Pirophosfat) Asam pantotenat, sbg bag. dr Koenzim A

FOSFORILASI OKSIDATIF

Electron Transport Complexes

4 multiprotein complexes in mitochondrial IM
NADH-CoQ (ubiquinone) oxidoreductase Succinate-CoQ oxidoreductase Ubiquinone-cytochrome c oxidoreductase Cytochrome c oxidase - reduction of O2

Contain a variety of prosthetic groups, iron-sulfur clusters Some subunits encoded by mitochondrial DNA

NADH-CoQ (ubiquinone) oxidoreductase (complex I)

2 electrons passed from NADH, through FMN, FeS intermediate electron carriers to ubiquinone (coenzyme Q) Ubiquinone - lipid soluble electron carrier Proton pumps transport 4 H+ from matrix to intermembrane space per pair of electrons Spatial organization important - protons used in reduction of ubiquinone come from matrix

Succinate-CoQ oxidoreductase (complex II)

Succinate-CoQ oxidoreductase
succinate dehydrogenase is a component No protons transported FAD, FeS serve as intermediate electron carriers

Ubiquinone-cytochrome c oxidoreductase (complex III)

Cytochrome c - peripheral protein, electron carrier Cytochromes can only accept 1 electron at a time, resulting in Q cycle 2 H+ from 1st Q deposited in intermembrane space, 1 e- to Cyt c, 1 e- to Qn 2 H+ from 2nd Q deposited in intermembrane space, 1 e- to Cyt c, 1 e- to Qn Qn with 2 e- takes 2 H+ from matrix.

Cytochrome c oxidase
catalyzes reduction of molecular oxygen 13 subunits Four protons translocated for each O2 reduced Accumulates 4 electrons (Cu+, Fe2+) for complete reduction before releasing products or toxic partially-reduced products O2 + 4 e- + 4 H+ --> 2 H2O occurs in matrix, thus removing 4 H+

ATP Synthase
ATP Synthase produces ATP from ADP & Pi H+ passage causes conformational changes (rotation) in F1, leading to release of ATP so ADP can bind again about 3 protons per ATP must pass through ATP synthase

The Big Picture

small molecule shuttles

molecules must be transported to and from matrix ATP-ADP translocase exports ATP, imports ADP movement of more negative ATP from matrix dissipates electrical potential across membrane, weakening gradient by 1 H+. Phosphate translocase uses 1 H+. cytosolic NADH DHAP is reduced by NADH to Glycerol-3-P in muscle Electrons passed through FAD to Q is less efficient, but allows transport against large NADH gradient

malate-aspartate shuttle
malate-aspartate shuttle used in heart, liver, kidney to transfer cytosolic reducing equivalents to matrix No loss in ATP generation (2.5 ATP per pair of electrons)

Malate Aspartate Shuttle

http://courses.cm.utexas.edu/emarcotte/ch339k/fall2005/Lecture-Ch19-2/Slide14.JPG

ATP yield/glucose
2 ATP - Glycolysis 3-5 ATP from 2 FADH from 2 NADH from glycolysis 5 ATP from 2 NADH from transition reaction 15 ATP from 6 NADH from TCA cycle 2 ATP from 2 GTP from TCA cycle 3 ATP from 2 FADH from TCA cycle 30-32 ATP from complete oxidation of glucose

Inhibitors
Electron flow can be inhibited by POISONS Useful in lab to control entry and exit points for electron transport studies Proton gradients are dissipated by DNP & FCCP, inhibiting ATP synthesis Thermogenin in brown adipose tissue dissipates proton gradient to generate heat

SEMOGA BERMANFAAT
YS 2010