title: A mutant of Escherichia coli fumarate reductase decoupled from electron transport.
abstract: The terminal electron-transfer enzyme fumarate reductase of Escherichia coli is a complex iron-sulfur flavoenzyme composed of four nonidentical subunits organized into two domains: FrdA and -B (a membrane-extrinsic catalytic domain) and FrdC and -D (a transmembrane anchor domain). We have identified a mutation within the membrane-intrinsic domain that alters the electron transfer properties of the iron-sulfur and flavin redox centers of the catalytic domain. Functional electron flow from the quinone analog 2,3-dimethyl-1,4-naphthoquinone or from the electron transport chain is impaired. However, the mutant enzyme can be reduced normally by single-electron donors such as the dye benzyl viologen. The mutant phenotype results from a single A----G transition changing His-82, within the second transmembrane alpha-helix of the FrdC anchor sequence, to an arginine. The mutation, physically located within the anchor domain, is manifested by altered catalytic properties, indicating that the intrinsic and extrinsic domains are conformationally connected. These results confirm the important role of the anchor subunits in functional electron transport and have implications for communication between intrinsic and extrinsic domains of membrane proteins.
mesh_terms: D001483:Base Sequence; D001594:Benzyl Viologen / Q000378:metabolism; D002384:Catalysis; D002462:Cell Membrane / Q000201:enzymology; D003001:Cloning, Molecular; D004578:Electron Spin Resonance Spectroscopy; D004579:Electron Transport; D004926:Escherichia coli / Q000201:enzymology / Q000235:genetics* / Q000378:metabolism; D005816:Genetic Complementation Test; D009154:Mutation; D009285:Naphthoquinones / Q000378:metabolism; D010084:Oxidation-Reduction; D010446:Peptide Fragments / Q000032:analysis; D013385:Succinate Dehydrogenase / Q000235:genetics*
chemical_list: D009285:Naphthoquinones; D010446:Peptide Fragments; D001594:Benzyl Viologen; C041498:2,3-dimethyl-1,4-naphthoquinone; D013385:Succinate Dehydrogenase