Flash-induced electron transfer through mitochondrial QH2: cytochrome c oxidoreductase in the presence of bacterial reaction centres and cytochrome c. Analysis of subsequent processes and effect of inhibitors.
In a system containing reaction centres isolated from Rhodopseudomonas sphaeroides mutant R26, and variable amounts of horse-heart cytochrome c and bovine-heart mitochondrial QH2: cytochrome c oxidoreductase in a medium containing 2 mM ascorbate and 0.1 microM phenazine methosulphate, electron transfer was induced by a single flash. Three distinct phases of electron transfer can be distinguished: the first event is the oxidation of cytochrome c, and this is followed by an equilibration between cytochrome c, cytochrome c1 and the Rieske [2Fe-2S] cluster. The actual rates of these processes depend on the concentrations of cytochrome c and the reductase. The slower third phase is the oxidation of ubiquinol, which can follow two pathways: one sensitive to antimycin and one sensitive to myxothiazole. The antimycin-sensitive pathway (t1/2 approximately equal to 10 ms) is an equilibration between the Q/QH2 couple and cytochrome b, but may also include a direct reduction of cytochrome b by the QB of the reaction centres. The myxothiazole-sensitive pathway is a coupled reduction of cytochrome b and the Rieske [2Fe-2S] cluster which rapidly equilibrates with cytochromes c1 and c. Both pathways are sensitive to 7-(n-heptadecyl)mercapto-6-hydroxy-5,8-quinoline quinone, but with different affinities. In the absence of inhibitors the initial reduction of cytochrome b (via both pathways) is followed by a net oxidation which is the resultant of a continuing reduction (together with the reduction of the Rieske [2Fe-2S] cluster) and an oxidation (via the antimycin-sensitive site) by quinone. The results are discussed in the light of linear and cyclic models proposed to explain electron transfer between cytochromes b and c. It is concluded that only the Q-cycle model fits the present experimental data.
D002734:Chlorophyll / Q000378:metabolism; D003574:Cytochrome c Group / Q000378:metabolism*; D003580:Cytochromes / Q000378:metabolism; D004579:Electron Transport; D003576:Electron Transport Complex IV / Q000378:metabolism*; D008928:Mitochondria / Q000378:metabolism*; D010084:Oxidation-Reduction; D012242:Rhodobacter sphaeroides; D013057:Spectrum Analysis; D014451:Ubiquinone / Q000378:metabolism*
D003574:Cytochrome c Group; D003580:Cytochromes; D014451:Ubiquinone; D002734:Chlorophyll; D003576:Electron Transport Complex IV