title: Heavy metal ions inhibit molybdoenzyme activity by binding to the dithiolene moiety of molybdopterin in Escherichia coli.
abstract: Molybdenum insertion into the dithiolene group on the 6-alkyl side-chain of molybdopterin is a highly specific process that is catalysed by the MoeA and MogA proteins in Escherichia coli. Ligation of molybdate to molybdopterin generates the molybdenum cofactor, which can be inserted directly into molybdoenzymes binding the molybdopterin form of the molybdenum cofactor, or is further modified in bacteria to form the dinucleotide form of the molybdenum cofactor. The ability of various metals to bind tightly to sulfur-rich sites raised the question of whether other metal ions could be inserted in place of molybdenum at the dithiolene moiety of molybdopterin in molybdoenzymes. We used the heterologous expression systems of human sulfite oxidase and Rhodobacter sphaeroides dimethylsulfoxide reductase in E. coli to study the incorporation of different metal ions into the molybdopterin site of these enzymes. From the added metal-containing compounds Na(2)MoO(4), Na(2)WO(4), NaVO(3), Cu(NO(3))(2), CdSO(4) and NaAsO(2) during the growth of E. coli, only molybdate and tungstate were specifically inserted into sulfite oxidase and dimethylsulfoxide reductase. Other metals, such as copper, cadmium and arsenite, were nonspecifically inserted into sulfite oxidase, but not into dimethylsulfoxide reductase. We showed that metal insertion into molybdopterin occurs beyond the step of molybdopterin synthase and is independent of MoeA and MogA proteins. Our study shows that the activity of molybdoenzymes, such as sulfite oxidase, is inhibited by high concentrations of heavy metals in the cell, which will help to further the understanding of metal toxicity in E. coli.
authors.affiliation: Department of Proteinanalytics, Institute of Biochemistry and Biology, University of Potsdam, Germany.
authors.forename: Meina
authors.initials: M
authors.lastname: Neumann
authors.forename: Silke
authors.initials: S
authors.lastname: Leimkühler
mesh_terms: D001665:Binding Sites; D003067:Coenzymes / Q000378:metabolism*; D004926:Escherichia coli / Q000378:metabolism*; D006801:Humans; D007506:Iron-Sulfur Proteins / Q000378:metabolism*; D008667:Metalloproteins / Q000378:metabolism*; D019216:Metals, Heavy / Q000737:chemistry / Q000378:metabolism*; D008982:Molybdenum / Q000378:metabolism; D000090006:Molybdenum Cofactors; D010088:Oxidoreductases / Q000378:metabolism*; D011621:Pteridines / Q000378:metabolism*; D012242:Rhodobacter sphaeroides / Q000201:enzymology; D050876:Sulfite Oxidase / Q000378:metabolism*; D013466:Sulfurtransferases / Q000378:metabolism*; D017973:Tungsten Compounds / Q000378:metabolism
chemical_list: D003067:Coenzymes; D007506:Iron-Sulfur Proteins; D008667:Metalloproteins; D019216:Metals, Heavy; D008982:Molybdenum; D000090006:Molybdenum Cofactors; D010088:Oxidoreductases; D011621:Pteridines; D050876:Sulfite Oxidase; D013466:Sulfurtransferases; D017973:Tungsten Compounds; C044659:molybdate; C022261:molybdopterin; C045664:dimethyl sulfoxide reductase; C085585:molybdopterin synthase; C045951:tungstate