Location of a cytochrome c binding site on the surface of flavocytochrome b(2)

D M Short, M D Walkinshaw, P Taylor, G A Reid, S K Chapman

Research output: Contribution to journalArticlepeer-review

Abstract

Saccharomyces cerevisiae flavocytochrome b(2) couples the oxidation of L-lactate to the reduction of cytochrome c. The second-order rate constant for cytochrome c reduction by flavocytochrome b(2) depends on the rate of complex formation and is sensitive to ionic strength, Mutations in the heme domain of flavocytochrome b(2) (Glu63-->Lys, Asp72-->Lps and the double mutation GluG3-->Lys:Asp72-->Lys) have significant effects on the reaction with cytochrome c, implicating these residues in complex formation. This kinetic information has been used to guide molecular modelling studies, which are consistent with there being no one single best-configuration. Rather, there is a set of possible complexes in which the docking-face of cytochrome c can approach flavocytochrome b(2) in a variety of orientations, Four cytochromes c can be accommodated on the flavocytochrome b(2) tetramer, with each cytochrome c forming interactions with only one flavocytochrome b(2) subunit. All the models involve residues 72 and 63 on flavocytochrome b(2) but in addition predict that GIu237 may also be important for complex formation. These acidic residues interact with the basic residues 13, 27 and 79 on cytochrome c, Through this triangle of interactions runs a possible sigma-tunnelling pathway for electron transfer. This pathway starts with the imidazole ring of His66 (a ligand to the heme-iron of flavocytochrome b(2)) and ends with the ring of Pro68, which is in van der Waals contact with the cytochrome c heme, In total, the edge-to-edge "through space" distance from the imidazole ring of His66 to the C3C pyrrole ring of cytochrome c is 13.1 Angstrom.

Original languageEnglish
Pages (from-to)246-252
Number of pages7
JournalJBIC Journal of Biological Inorganic Chemistry
Volume3
Issue number3
Publication statusPublished - Jun 1998

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