Energy transduction by the reconstituted b-c1 complex from yeast mitochondria. Inhibitory effects of dicyclohexylcarbodiimide.
A purified cytochrome b-c1 complex isolated from yeast mitochondria has been reconstituted into proteoliposomes. The reconstituted comp]lex catalyzed antimycin A-sensitive electron transfer from different analogues of coenzyme Q to cytochrome c. The reconstituted complex was also capable of energy conservation as indicated by uncoupler-stimulated rates of electron transfer, electrogenic proton ejection, and reversed electron flow from cytochrome b to coenzyme Q2 in the presence of antimycin A driven by a valinomycin-induced K+-diffusion potential (negative inside). Close to four protons were ejected per two electrons transported through the reconstituted b-c1 complex with ferricyanide as an artificial and impermeable electron acceptor.l The H+/2e- ratio decreased to two in the presence of the proton-conducting agent, carbonyl cyanide m-chlorophenylhydrazone. The same processes were studied in parallel in energy-conserving site 2 of rat liver mitochondria with similar results. In the reconstituted b-c1 complex, dicyclohexylcarbodiimide (DCCD) blocked the function of the electrogenic proton translocating device in the forward direction of proton ejection as well as in the backwards direction, measured as reversed electron flow from cytochrome b to coenzyme Q2 driven by a K+-diffusion potential. The primary effect of DCCD is localized on the proton ejection process, as the low proton conductance of the proteoliposome membrane was totally preserved after DCCD treatment.
D000818:Animals; D002234:Carbodiimides / Q000494:pharmacology*; D003573:Cytochrome b Group / Q000378:metabolism; D004024:Dicyclohexylcarbodiimide / Q000494:pharmacology*; D004579:Electron Transport; D014450:Electron Transport Complex III; D006863:Hydrogen-Ion Concentration; D007700:Kinetics; D008081:Liposomes; D008928:Mitochondria / Q000201:enzymology*; D008930:Mitochondria, Liver / Q000378:metabolism; D009097:Multienzyme Complexes / Q000037:antagonists & inhibitors*; D009247:NADH, NADPH Oxidoreductases / Q000037:antagonists & inhibitors*; D010101:Oxygen Consumption; D010743:Phospholipids / Q000494:pharmacology; D011808:Quinone Reductases / Q000037:antagonists & inhibitors*; D051381:Rats; D012441:Saccharomyces cerevisiae / Q000201:enzymology*
D002234:Carbodiimides; D003573:Cytochrome b Group; D008081:Liposomes; D009097:Multienzyme Complexes; D010743:Phospholipids; D004024:Dicyclohexylcarbodiimide; D009247:NADH, NADPH Oxidoreductases; D011808:Quinone Reductases; D014450:Electron Transport Complex III