title: Catalytically active monoamine oxidase type A from human liver expressed in Saccharomyces cerevisiae contains covalent FAD.
abstract: Monoamine oxidase type A from human liver cDNA was expressed in Saccharomyces cerevisiae. This enzyme's properties with respect to Km and Ki values for kynuramine and amphetamine, respectively, were similar to values for human placental enzyme. As expected, clorgyline inhibited the yeast enzyme at lower concentrations than deprenyl. Interestingly, the FAD cofactor was covalently attached and fluorescence properties of the enzyme bound prosthetic group indicate that it is attached to a cysteine residue, the same linkage observed in other monoamine oxidases. The yield of expressed enzyme is about 15 mg/l of culture with an A600 of 15. It is suggested that covalent flavin attachment proceeds by an autoflavination mechanism.
authors: Department of Veterans Affairs Medical Center, Molecular Biology, San Francisco, CA 94121.
authors: W
authors: W
authors: Weyler
authors: C C
authors: CC
authors: Titlow
authors: J I
authors: JI
authors: Salach
mesh_terms: D001483:Base Sequence; D003001:Cloning, Molecular; D003010:Clorgyline / Q000494:pharmacology; D004247:DNA / Q000737:chemistry; D004274:DNA, Recombinant; D005182:Flavin-Adenine Dinucleotide / Q000737:chemistry*; D005453:Fluorescence; D006801:Humans; D007700:Kinetics; D008099:Liver / Q000201:enzymology*; D008969:Molecular Sequence Data; D008995:Monoamine Oxidase / Q000096:biosynthesis / Q000235:genetics*; D010920:Placenta / Q000201:enzymology; D015183:Restriction Mapping; D012441:Saccharomyces cerevisiae / Q000235:genetics*; D012642:Selegiline / Q000494:pharmacology
chemical_list: D004274:DNA, Recombinant; D005182:Flavin-Adenine Dinucleotide; D012642:Selegiline; D004247:DNA; D008995:Monoamine Oxidase; D003010:Clorgyline