title: Escherichia coli K-12 copper-containing monoamine oxidase: investigation of the copper binding ligands by site-directed mutagenesis, elemental analysis and topa quinone formation.
abstract: Copper-containing monoamine oxidase (EC 1.4.3.6) from Escherichia coli is a periplasmic enzyme containing topa quinone in addition to divalent copper as a cofactor. The amino acid sequence of E. coli monoamine oxidase was compared to several cloned amine oxidase genes and five well-conserved histidine residues were found. Site-directed mutagenesis studies were performed to determine which histidine residue serves as the binding ligand to the copper. Enzyme activity, absorption spectrum, and atomic absorption spectrophotometry of the mutant enzymes indicated that histidines 470, 554, and 556 are the copper binding ligands. The absorption spectra of phenylhydrazine derivatives suggested that copper is necessary for topa quinone formation.
authors.affiliation: Department of Food Science and Technology, Faculty of Agriculture, Kyoto University, Japan.
authors.forename: J H
authors.initials: JH
authors.lastname: Roh
authors.forename: Y
authors.initials: Y
authors.lastname: Takenaka
authors.forename: H
authors.initials: H
authors.lastname: Suzuki
authors.forename: K
authors.initials: K
authors.lastname: Yamamoto
authors.forename: H
authors.initials: H
authors.lastname: Kumagai
mesh_terms: D000595:Amino Acid Sequence; D001483:Base Sequence; D001665:Binding Sites / Q000235:genetics; D003001:Cloning, Molecular; D003300:Copper / Q000737:chemistry; D017931:DNA Primers / Q000235:genetics; D004269:DNA, Bacterial / Q000235:genetics; D004295:Dihydroxyphenylalanine / Q000031:analogs & derivatives / Q000096:biosynthesis; D004602:Elements; D004926:Escherichia coli / Q000201:enzymology* / Q000235:genetics; D006639:Histidine / Q000737:chemistry / Q000235:genetics; D008024:Ligands; D008969:Molecular Sequence Data; D008995:Monoamine Oxidase / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D016297:Mutagenesis, Site-Directed; D017386:Sequence Homology, Amino Acid; D013053:Spectrophotometry
chemical_list: D017931:DNA Primers; D004269:DNA, Bacterial; D004602:Elements; D008024:Ligands; D006639:Histidine; D004295:Dihydroxyphenylalanine; C067626:6-hydroxydopa quinone; D003300:Copper; D008995:Monoamine Oxidase