The enzymes of the bc1 complex family (ubiquinol:cytochrome c oxidoreductases, and the closely related b6f complex of oxygenic photosynthesis), carry the energy flux of the biosphere, serving as the central enzymes of respiratory and photosynthetic electron transfer chains.
The aim of this project has been to understand how these important enzymes function. X-ray crystallographic structures for several mitochondrial complexes have recently been solved in collaboration with Dr. Ed Berry, and an extensive analysis in the light of previous work has provide new insights on function. Much biophysical work has established the basic mechanism, and the focus in recent years has been on putting this into a structural context. The complexes catalyze the oxidation of ubiquinol and the reduction of cytochrome c through a modified Q-cycle. Three catalytic subunits, a cytochrome b with two hemes, cytochrome c1 and an iron sulfur protein, house the mechanism. These are well conserved across the bacterial/mitochondrial divide. Two separate internal electron transfer chains connect three catalytic sites that catalyze oxidation and reduction of the quinone pool, and reduction of cytochrome c. Electron transfer across the membrane, and coupling of these redox reactions to the release or uptake of protons, allows the complex to generate the transmembrane gradient that drives ATP synthesis. From our analysis of the structure, we have suggested some novel extensions of this basic mechanism, including a dramatic movement of the iron sulfur protein between its two reaction partners, a revised mechanism for the reaction by which quinol is oxidized, and a more detailed understanding of the quinone reduction site. In the renewal period, we will make use of the structure in an extended exploration of the molecular mechanism, using spectroscopic methods, and biophysical, molecular engineering and biochemical protocols developed under the grant. Apart from its intrinsic interest, the bc1 complex is a major site of production of oxygen radicals, which cause cell aging and DNA damage leading to cancer, and also the locus of inherited genetic diseases. Natural inhibitors block turnover by mimicking quinone at the catalytic sites, and commercial interest has centered on the possibility of using these as green pesticides.
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