title: The Saccharomyces cerevisiae succinate dehydrogenase does not require heme for ubiquinone reduction.
abstract: The coupling of succinate oxidation to the reduction of ubiquinone by succinate dehydrogenase (SDH) constitutes a pivotal reaction in the aerobic generation of energy. In Saccharomyces cerevisiae, SDH is a tetramer composed of a catalytic dimer comprising a flavoprotein subunit, Sdh1p and an iron-sulfur protein, Sdh2p and a heme b-containing membrane-anchoring dimer comprising the Sdh3p and Sdh4p subunits. In order to investigate the role of heme in SDH catalysis, we constructed an S. cerevisiae strain expressing a mutant enzyme lacking the two heme axial ligands, Sdh3p His-106 and Sdh4p Cys-78. The mutant enzyme was characterized for growth on a non-fermentable carbon source, for enzyme assembly, for succinate-dependent quinone reduction and for its heme b content. Replacement of both Sdh3p His-106 and Sdh4p Cys-78 with alanine residues leads to an undetectable level of cytochrome b(562). Although enzyme assembly is slightly impaired, the apocytochrome SDH retains a significant ability to reduce quinone. The enzyme has a reduced affinity for quinone and its catalytic efficiency is reduced by an order of magnitude. To better understand the effects of the mutations, we employed atomistic molecular dynamic simulations to investigate the enzyme's structure and stability in the absence of heme. Our results strongly suggest that heme is not required for electron transport from succinate to quinone nor is it necessary for assembly of the S. cerevisiae SDH.
authors.affiliation: Department of Biochemistry, University of Alberta, Edmonton, Alberta, Canada T6G 2H7.
authors.forename: Kayode S
authors.initials: KS
authors.lastname: Oyedotun
authors.forename: Clarissa S
authors.initials: CS
authors.lastname: Sit
authors.forename: Bernard D
authors.initials: BD
authors.lastname: Lemire
mesh_terms: D019943:Amino Acid Substitution / Q000235:genetics; D004579:Electron Transport / Q000235:genetics; D006418:Heme / Q000502:physiology; D010084:Oxidation-Reduction; D012441:Saccharomyces cerevisiae / Q000201:enzymology* / Q000235:genetics; D029701:Saccharomyces cerevisiae Proteins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D013056:Spectrophotometry, Ultraviolet; D013385:Succinate Dehydrogenase / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D014451:Ubiquinone / Q000378:metabolism*
chemical_list: D029701:Saccharomyces cerevisiae Proteins; D014451:Ubiquinone; D006418:Heme; D013385:Succinate Dehydrogenase