Voltammetry of a flavocytochrome c3:The lowest potential heme modulates fumarate reduction rates

Butt, Julea N. ORCID: https://orcid.org/0000-0002-9624-5226, Thornton, Jeremy, Richardson, David J. ORCID: https://orcid.org/0000-0002-6847-1832 and Dobbin, Paul S. (2000) Voltammetry of a flavocytochrome c3:The lowest potential heme modulates fumarate reduction rates. Biophysical Journal, 78 (2). pp. 1001-1009. ISSN 0006-3495

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Iron-induced flavocytochrome C3, IfC3, from Shewanella frigidimarina NCIMB400, derivatized with a 2-pyridyl disulfide label, self-assembles on gold electrodes as a functional array whose fumarate reductase activity as viewed by direct electrochemistry is indistinguishable from that of IfC3 adsorbed on gold or graphite electrodes. The enhanced stability of the labeled protein's array permits analysis at a rotating electrode and limiting catalytic currents fit well to a Michaelis-Menten description of enzyme kinetics with K(M) = 56 ± 20 μM, pH 7.5, comparable to that obtained in solution assays. At fumarate concentrations above 145 μM cyclic voltammetry shows the catalytic response to contain two features. The position and width of the lower potential component centered on -290 mV and corresponding to a one-electron wave implicates the oxidation state of the lowest potential heme of IfC3 as a defining feature in the mechanism of fumarate reduction at high turnover rates. We propose the operation of dual pathways for electron transfer to the active site of IfC3 with the lowest potential heme acting as an electron relay on one of these pathways.

Item Type: Article
Additional Information: Funding Information: D. J. R. and P. S. D. also thank the Wellcome Trust for funding by project grant 046547. Funding Information: J. N. B. gratefully acknowledges the Wellcome Trust for a Research Career Development Fellowship, Grant 050709.
Uncontrolled Keywords: biophysics ,/dk/atira/pure/subjectarea/asjc/1300/1304
Faculty \ School: Faculty of Science > School of Chemical Sciences and Pharmacy (former - to 2009)
Faculty of Science > School of Biological Sciences
UEA Research Groups: Faculty of Science > Research Groups > Organisms and the Environment
Faculty of Science > Research Groups > Molecular Microbiology
Faculty of Science > Research Centres > Centre for Molecular and Structural Biochemistry
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Depositing User: LivePure Connector
Date Deposited: 14 Jul 2022 16:31
Last Modified: 15 May 2023 00:53
URI: https://ueaeprints.uea.ac.uk/id/eprint/86126
DOI: 10.1016/S0006-3495(00)76658-6

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