title: Identification of a lysine residue important for the catalytic activity of yeast farnesyl diphosphate synthase.
abstract: The Saccharomyces cerevisiae ERG20 gene (encoding farnesyl diphosphate synthase) has been subjected to a set of mutations at the catalytic site, at position K254 to determine the in vivo impact. The mutated strains have been shown to exhibit various growth rates, sterol profiles and monoterpenol producing capacities. The results obtained suggest that K at position 254 helps to stabilize one of the three Mg(2+) forming a bridge between the enzyme and DMAPP, and demonstrate that destabilizing two of the three Mg(2+) ions, by introducing a double mutation at positions K197 and K254, results in a loss of FPPS activity and a lethal phenotype.
authors.affiliation: Laboratoire Métabolisme Secondaire de la Vigne, Univ. Strasbourg, INRA, Inst. Natl. Recherche Agron., Métab. Second. Vigne, Unit Mixte Recherche Santé Vigne and Qual. Vins, Colmar, France. fischer@colmar.inra.fr
authors.forename: Marc J C
authors.initials: MJ
authors.lastname: Fischer
authors.forename: Sophie
authors.initials: S
authors.lastname: Meyer
authors.forename: Patricia
authors.initials: P
authors.lastname: Claudel
authors.forename: Marc
authors.initials: M
authors.lastname: Bergdoll
authors.forename: Francis
authors.initials: F
authors.lastname: Karst
mesh_terms: D000595:Amino Acid Sequence; D020134:Catalytic Domain; D048708:Cell Growth Processes / Q000235:genetics; D002470:Cell Survival / Q000235:genetics; D004795:Enzyme Stability; D051228:Geranyltranstransferase / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D049452:Green Fluorescent Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D045782:Hemiterpenes; D008239:Lysine / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D008958:Models, Molecular; D008969:Molecular Sequence Data; D039821:Monoterpenes / Q000378:metabolism; D016297:Mutagenesis, Site-Directed; D009943:Organophosphorus Compounds; D010840:Phytosterols / Q000378:metabolism; D011993:Recombinant Fusion Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D012441:Saccharomyces cerevisiae / Q000201:enzymology* / Q000235:genetics; D029701:Saccharomyces cerevisiae Proteins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D016415:Sequence Alignment
chemical_list: D045782:Hemiterpenes; D039821:Monoterpenes; D009943:Organophosphorus Compounds; D010840:Phytosterols; D011993:Recombinant Fusion Proteins; D029701:Saccharomyces cerevisiae Proteins; D049452:Green Fluorescent Proteins; C043060:3,3-dimethylallyl pyrophosphate; D051228:Geranyltranstransferase; D008239:Lysine