Mechanism of ubiquinol oxidation by the bc1 complex: Different domains of the quinol binding pocket and their role in the mechanism and binding of inhibitors

Antony R. Crofts, Blanca Barquera, Robert B. Gennis, Richard Kuras, Mariana Guergova-Kuras, Edward A. Berry

Research output: Contribution to journalArticlepeer-review


Structures of mitochondrial ubihydroquinone:cytochrome c oxidoreductase (bc1 complex) from several animal sources have provided a basis for understanding the functional mechanism at the molecular level. Using structures of the chicken complex with and without inhibitors, we analyze the effects of mutation on quinol oxidation at the Q(o) site of the complex. We suggest a mechanism for the reaction that incorporates two features revealed by the structures, a movement of the iron sulfur protein between two separate reaction domains on cytochrome c1 and cytochrome b and a bifurcated volume for the Q(o) site. The volume identified by inhibitor binding as the Q(o) site has two domains in which inhibitors of different classes bind differentially; a domain proximal to heme b(L), where myxothiazole and β- methoxyacrylate(MOA-) type inhibitors bind (class II), and a distal domain close to the iron sulfur protein docking interface, where stigmatellin and 5- n-undecyl-6-hydroxy-4,7-dioxobenzothiaole (UHDBT) bind (class I). Displacement of one class of inhibitor by another is accounted for by the overlap of their volumes, since the exit tunnel to the lipid phase forces the hydrophobic 'tails' to occupy common space. We conclude that the site can contain only one 'tailed' occupant, either an inhibitor or a quinol or one of their reaction products. The differential sensitivity of strains with mutations in the different domains is explained by the proximity of the affected residues to the binding domains of the inhibitors. New insights into mechanism are provided by analysis of mutations that affect changes in the electron paramagnetic resonance (EPR) spectrum of the iron sulfur protein, associated with its interactions with the Q(o)-site occupant. The structures show that all interactions with the iron sulfur protein must occur at the distal position. These include interactions between quinone, or class I inhibitors, and the reduced iron sulfur protein and formation of a reaction complex between quinol and oxidized iron sulfur protein. The step with high activation energy is after formation of the reaction complex, likely in formation of the semiquinone and subsequent dissociation of the complex into products. We suggest that further progress of the reaction requires a movement of semiquinone to the proximal position, thus mapping the bifurcated reaction to the bifurcated volume. We suggest that such a movement, together with a change in conformation of the site, would remove any semiquinone formed from further interaction with the oxidized [2Fe-2S] center and also from reaction with O2 to form superoxide anion. We also identify two separate reaction paths for exit of the two protons released in quinol oxidation.

Original languageEnglish (US)
Pages (from-to)15807-15826
Number of pages20
Issue number48
StatePublished - Nov 30 1999

ASJC Scopus subject areas

  • Biochemistry


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