title: MoaA of Arthrobacter nicotinovorans pAO1 involved in Mo-pterin cofactor synthesis is an Fe-S protein.
abstract: MoaA, involved in an early step in the biosynthesis of the molybdopterin cofactor (MoCo), has not yet been characterized biochemically and the reaction it catalyzes is unknown. We overexpressed MoaA from pAO1 of Arthrobacter nicotinovorans in Escherichia coli as a N-terminal fusion with either glutathione-S-transferase or a 6-histidine tag. The pAO1 encoded MoaA as well as the fusion proteins functionally complement E. coli moaA mutants. Here we show that purified MoaA contains approximately 4 microM Fe and approximately 3 microM acid-labile S/microM protein. EPR spectroscopy revealed a predominant signal at g(av) = 2.01, indicative of a [3Fe-xS] cluster.
authors.affiliation: Institut für Biochemie und Molekularbiologie, Freiburg, Germany.
authors.forename: C
authors.initials: C
authors.lastname: Menéndez
authors.forename: D
authors.initials: D
authors.lastname: Siebert
authors.forename: R
authors.initials: R
authors.lastname: Brandsch
mesh_terms: D000595:Amino Acid Sequence; D001173:Arthrobacter / Q000378:metabolism*; D001483:Base Sequence; D003067:Coenzymes / Q000096:biosynthesis; D017931:DNA Primers; D004578:Electron Spin Resonance Spectroscopy; D005982:Glutathione Transferase / Q000096:biosynthesis; D007506:Iron-Sulfur Proteins / Q000378:metabolism*; D008667:Metalloproteins / Q000737:chemistry / Q000378:metabolism*; D008969:Molecular Sequence Data; D008982:Molybdenum / Q000378:metabolism*; D000090006:Molybdenum Cofactors; D016297:Mutagenesis, Site-Directed; D011621:Pteridines / Q000737:chemistry / Q000378:metabolism*; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis / Q000737:chemistry / Q000378:metabolism; D016324:Sequence Tagged Sites
chemical_list: D003067:Coenzymes; D017931:DNA Primers; D005982:Glutathione Transferase; D007506:Iron-Sulfur Proteins; D008667:Metalloproteins; D008982:Molybdenum; D000090006:Molybdenum Cofactors; D011621:Pteridines; D011993:Recombinant Fusion Proteins; C022261:molybdopterin