Modification of the catalytic function of the mitochondrial cytochrome b-c1 complex by dicyclohexylcarbodiimide.
N,N'-Dicyclohexylcarbodiimide (DCCD) induces a complex set of effects on the succinate-cytochrome c span of the mitochondrial respiratory chain. At concentrations below 1000 mol per mol of cytochrome c1, DCCD is able to block the proton-translocating activity associated to succinate or ubiquinol oxidation without inhibiting the steady-state redox activity of the b-c1 complex either in intact mitochondrial particles or in the isolated ubiquinol-cytochrome c reductase reconstituted in phospholipid vesicles. In parallel to this, DCCD modifies the redox responses of the endogenous cytochrome b, which becomes more rapidly reduced by succinate, and more slowly oxidized when previously reduced by substrates. At similar concentrations the inhibitor apparently stimulates the redox activity of the succinate-ubiquinone reductase. Moreover, DCCD, at concentrations about one order of magnitude higher than those blocking proton translocation, produces inactivation of the redox function of the b-c1 complex. The binding of [14C]DCCD to the isolated b-c1 complex has shown that under conditions leading to the inhibition of the proton-translocating activity of the enzyme, a subunit of about 9500 Da, namely Band VIII, is the most heavily labelled polypeptide of the complex. The possible correlations between the various effects of DCCD and its modification of the b-c1 complex are discussed.
D000818:Animals; D002234:Carbodiimides / Q000494:pharmacology*; D002417:Cattle; D003574:Cytochrome c Group / Q000378:metabolism; D004024:Dicyclohexylcarbodiimide / Q000494:pharmacology*; D014450:Electron Transport Complex III; D007700:Kinetics; D008564:Membrane Potentials / Q000187:drug effects; D008929:Mitochondria, Heart / Q000201:enzymology*; D008930:Mitochondria, Liver / Q000201:enzymology*; D009097:Multienzyme Complexes / Q000378:metabolism*; D009247:NADH, NADPH Oxidoreductases / Q000378:metabolism*; D010100:Oxygen / Q000378:metabolism; D011808:Quinone Reductases / Q000378:metabolism*; D051381:Rats; D013385:Succinate Dehydrogenase / Q000378:metabolism; D014451:Ubiquinone / Q000378:metabolism
D002234:Carbodiimides; D003574:Cytochrome c Group; D009097:Multienzyme Complexes; D014451:Ubiquinone; D004024:Dicyclohexylcarbodiimide; D013385:Succinate Dehydrogenase; D009247:NADH, NADPH Oxidoreductases; D011808:Quinone Reductases; D014450:Electron Transport Complex III; D010100:Oxygen