Dimeric ubiquinol:cytochrome c reductase of Neurospora mitochondria contains one cooperative ubiquinone-reduction centre.
Dimeric ubiquinol:cytochrome c reductase of Neurospora mitochondria was isolated as a protein-Triton complex and free of ubiquinol (Q). The enzyme was incorporated into phosphatidylcholine membranes together with Q. The effects of varying the molar ratio of Q to enzyme on the electron transfer from duroquinol (DHQ2) to the cytochromes c, c1 and b were studied. The rate of electron flow from DQH2 to cytochrome c was 15 times increased by Q and was maximal when one molecule of Q was bound to one enzyme dimer. The apparent Km value for DQH2 of the Q-free enzyme was 5 microM and of the Q-supplemented enzyme 25 microM. The pre-steady-state rate of electron transfer from DQH2 to cytochrome c1 was also 15 times increased by Q and was maximal with one Q molecule bound to one enzyme dimer. This effect of Q was inhibited by antimycin. The pre-steady-state rate of electron transfer from DQH2 to cytochrome b was 5 times decreased when Q was bound to the enzyme and this effect of Q was insensitive to myxothiazol. The H+/2e- stoichiometry with DQH2 as substrate of the Q-supplemented enzyme was 3.6. These results are interpreted in accordance with a Q-cycle mechanism operating in a dimeric cytochrome reductase. Each enzyme monomer catalyses a single electron transfer from the QH2-oxidation centre to the Q-reduction centre and the two monomers cooperate in the reduction of Q to QH2 at one Q-reduction centre. This centre contains two different binding sites for Q. DQH2 does not properly react at the QH2-oxidation centre. DQH2, however, binds to the loose Q-binding site of the Q-reduction centre and reduces the Q bound to the tight Q-binding site of the centre. The QH2 thus formed at the Q-reduction centre serves as electron donor for the QH2-oxidation centre.
D001692:Biological Transport; D003574:Cytochrome c Group / Q000378:metabolism; D004579:Electron Transport; D014450:Electron Transport Complex III; D006873:Hydroquinones / Q000378:metabolism; D008928:Mitochondria / Q000201:enzymology*; D009097:Multienzyme Complexes / Q000032:analysis*; D009491:Neurospora / Q000201:enzymology*; D010084:Oxidation-Reduction; D011522:Protons; D011808:Quinone Reductases / Q000032:analysis*; D014451:Ubiquinone / Q000032:analysis / Q000378:metabolism*
D003574:Cytochrome c Group; D006873:Hydroquinones; D009097:Multienzyme Complexes; D011522:Protons; D014451:Ubiquinone; D011808:Quinone Reductases; D014450:Electron Transport Complex III; C025204:Ubiquinone Q2; C016421:duroquinol