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F_Lec_18_color - (90 micromoles 18 umoles 30 umoles 45 umoles ADP=90 umoles Interpretation of Results a β-OH-butyrate Conversion of 90 umol ADP(or

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1 Use: Inhibitors of electron transport Rotenone/Amytal - inhibits complex I Antimycin - inhibits complex III CN - inhibits complex IV Where do electrons enter the ETC? II Rotenone/ amytal Antimycin a CN
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2 II FADH 2 FAD TMPD Ascorbate
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3 II FADH 2 FAD TMPD Ascorbate
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4 The Oxidation of NADH or FADH 2 by O 2 is Tightly Coupled to the Phosphorylation of ADP 2 NADH 1 O 2 nADP + nP i nATP How many moles of ATP are synthesized from the reduction of 1 mole O 2 ? • Measure the amount of O 2 consumed (reduced to 2H 2 O) for any given amount of ADP added. • Experimental Conditions - same as the inhibitor expt (no ADP initially, excess PO 4 ) - isolated mito’s in buffer containing excess phosphate - addition of ADP + an electron donor starts electron transport ADP, must be limiting! NADH, Pi, O 2 must be in excess.
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5 O 2 Consumption as a function of ADP P’n Conditions: Isolated mitochondria in buffer containing excess PO 4 . Reaction is initiated by addition of ADP and e- donor.
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Unformatted text preview: (90 micromoles) 18 umoles 30 umoles 45 umoles ADP=90 umoles Interpretation of Results a) β-OH-butyrate Conversion of 90 umol ADP (or PO 4 ) Æ ATP requires 18 umol O 2 (36 umol O) P/O = 90/36 = 2.5 b) Succinate Conversion of 90 umol ADP (or PO 4 ) Æ ATP requires 30 umol O 2 (60 umol O) P/O = 90/60 = 1.5 c) TMPD/Ascorbate P/O = 90/90 = 1 6 How many protons are pumped for every electron pair? 7 II FADH 2 FAD TMPD Ascorbate Complex I Lehninger Fig 19-9 8 Four Types of Redox Centers in the Electron Transport Chain 1. Flavins – FMN, FAD 2. Iron – Sulfur Centers [2Fe–2S], [4Fe-4S] 3. Hemes 4. Cu Ions Iron-Sulfur Centers 9 Complex I Lehninger Fig 19-9 10 • • Complex II Q • Oxygen is a bi-radical • Can accept e-s only 1 at a time • ETC starts with e-pairs …. • O 2 •...
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This note was uploaded on 04/16/2009 for the course MCDB 108b taught by Professor Christoffersen during the Winter '08 term at UCSB.

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F_Lec_18_color - (90 micromoles 18 umoles 30 umoles 45 umoles ADP=90 umoles Interpretation of Results a β-OH-butyrate Conversion of 90 umol ADP(or

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