title: EPR studies of wild-type and several mutants of cytochrome c oxidase from Rhodobacter sphaeroides: Glu286 is not a bridging ligand in the cytochrome a3-CuB center.
abstract: Wild-type and several mutants of cytochrome c oxidase from Rhodobacter sphaeroides were characterized by EPR spectroscopy. A pH-induced g12 signal, seen previously in mammalian cytochrome oxidase and assigned to the presence of a bridging carboxyl ligand in the bimetallic cytochrome a3-CuB site, is found also in the bacterial enzyme. Mutation of glutamate-286 to glutamine inactivates the enzyme but does not affect this signal, demonstrating that the carboxyl group of this residue is not the bridging ligand. Three mutants, M106Q, located one helix turn below a histidine ligand to cytochrome a, and T352A as well as F391Q, located close to the bimetallic center, are shown to affect dramatically the low-spin heme signal of cytochrome a. These mutants are essentially inactive, suggesting that these three mutations result in alterations to cytochrome a that render the oxidase non-functional.
authors.affiliation: School of Chemical Sciences, University of Illinois, Urbana 61801, USA.
authors.forename: D M
authors.initials: DM
authors.lastname: Mitchell
authors.forename: R
authors.initials: R
authors.lastname: Aasa
authors.forename: P
authors.initials: P
authors.lastname: Adelroth
authors.forename: P
authors.initials: P
authors.lastname: Brzezinski
authors.forename: R B
authors.initials: RB
authors.lastname: Gennis
authors.forename: B G
authors.initials: BG
authors.lastname: Malmström
mesh_terms: D001483:Base Sequence; D003300:Copper / Q000737:chemistry*; D017936:Cytochrome a Group / Q000378:metabolism; D004578:Electron Spin Resonance Spectroscopy; D003576:Electron Transport Complex IV / Q000737:chemistry* / Q000235:genetics; D018698:Glutamic Acid / Q000737:chemistry* / Q000235:genetics; D005973:Glutamine / Q000737:chemistry; D006639:Histidine; D006863:Hydrogen-Ion Concentration; D008969:Molecular Sequence Data; D016297:Mutagenesis, Site-Directed; D017433:Protein Structure, Secondary; D012242:Rhodobacter sphaeroides / Q000201:enzymology*
chemical_list: D017936:Cytochrome a Group; D005973:Glutamine; D018698:Glutamic Acid; D006639:Histidine; D003300:Copper; D003576:Electron Transport Complex IV