The pathway of electron flow through ubiquinol:cytochrome c oxidoreductase in the respiratory chain. Evidence from inhibition studies for a modified 'Q cycle'.

1. Cytochrome spectra of the liver and heart mitochondria incubated under various conditions are presented to compare the effects of antimycin, colletotrichin and 2-heptyl-4-hydroxyquinoline N-oxide (HQNO) additions. 2. Under aerobic conditions, in State 4, in the presence of uncoupler or in the presence of cyanide, all three inhibitors caused oxidation of cytochromes c and c1, but different changes in the spectra of the b cytochromes. Antimycin caused oxidation of a peak at 558 nm and reduction of peaks at 562 nm and 566 nm, whereas colletotrichin caused reduction of peaks at 558 nm and 566 nm and oxidation at 562 nm. HQNO had an effect on the spectra intermediate between those of the two other inhibitors. 3. Under aerobic conditions in the presence of 5 mM-succinate and 5 mM-fumarate, antimycin caused reduction of a peak at 566 nm and oxidation of a peak at 558 nm, whereas colletotrichin had the reverse effect and HQNO caused reduction of a peak at 562 nm. 4. Colletotrichin inhibition of the ADP-stimulated oxidation of glutamate + malate was enhanced by succinate addition and declined again with rotenone addition. Similar but smaller effects were seen with inhibition by antimycin and HQNO. 5. Cytochrome spectra are shown of the effects of ADP and uncoupler addition to stimulate respiration progressively. 6. The results are interpreted in terms of a modified 'Q cycle' [Mitchell (1976) J. Theor. Biol. 62, 327-367] in which the three inhibitors are postulated to displace ubiquinone and ubisemiquinone specifically bound to cytochromes b on both sides of the membrane. 7. It is suggested that cytochromes b558 and b566 are the same b cytochrome located on the outer surface of the membrane, but binding ubisemiquinone or colletotrichin and ubiquinone or antimycin respectively. Cytochrome b562 is postulated to be on the inner surface of the mitochondrial membrane and to bind either ubiquinone or ubisemiquinone, HQNO would bind to the reduced form of the cytochrome and colletotrichin to the oxidized form. 8. Sites for the locus of action of glucagon and the protonmotive force on electron flow are suggested.