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

Duncan M Short, Malcolm D Walkinshaw, Paul Taylor, Graeme A Reid, Stephen K Chapman

    Research output: Contribution to journalArticle

    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
    JournalJournal of Biological Inorganic Chemistry
    Volume3
    Issue number3
    DOIs
    Publication statusPublished - Jun 1998

    Fingerprint

    Cytochromes c
    Binding Sites
    Heme
    cytochrome b558
    Pyrroles
    Molecular modeling
    Ionic strength
    Yeast
    Lactic Acid
    Rate constants
    Iron
    Ligands
    Oxidation
    Kinetics
    Electrons

    Keywords

    • flavocytochrome b(2)
    • B2
    • COMPLEX
    • FLAVIN
    • PROTEIN
    • electron transfer
    • B(5)
    • INTERDOMAIN HINGE
    • site-directed mutagenesis
    • ELECTRON-TRANSFER-REACTION
    • cytochrome c
    • modelling

    Cite this

    Short, D. M., Walkinshaw, M. D., Taylor, P., Reid, G. A., & Chapman, S. K. (1998). Location of a cytochrome c binding site on the surface of flavocytochrome b(2). Journal of Biological Inorganic Chemistry, 3(3), 246-252. https://doi.org/10.1007/PL00010651
    Short, Duncan M ; Walkinshaw, Malcolm D ; Taylor, Paul ; Reid, Graeme A ; Chapman, Stephen K. / Location of a cytochrome c binding site on the surface of flavocytochrome b(2). In: Journal of Biological Inorganic Chemistry. 1998 ; Vol. 3, No. 3. pp. 246-252.
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    Location of a cytochrome c binding site on the surface of flavocytochrome b(2). / Short, Duncan M ; Walkinshaw, Malcolm D; Taylor, Paul; Reid, Graeme A; Chapman, Stephen K.

    In: Journal of Biological Inorganic Chemistry, Vol. 3, No. 3, 06.1998, p. 246-252.

    Research output: Contribution to journalArticle

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    AU - Short, Duncan M

    AU - Walkinshaw, Malcolm D

    AU - Taylor, Paul

    AU - Reid, Graeme A

    AU - Chapman, Stephen K

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    AB - 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.

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    KW - INTERDOMAIN HINGE

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