title: Effect of site-specific mutagenesis of tyrosine-55, methionine-110 and histidine-217 in porcine kidney D-amino acid oxidase on its catalytic function.
abstract: To assess the contributions of Tyr-55, Met-110 and His-217 in porcine kidney D-amino acid oxidase (EC 1.4.3.3, DAO) to its catalytic function, we constructed three mutant cDNAs coding for the enzymes possessing Phe-55, Leu-110 and Leu-217 by site-specific mutagenesis. The mutant and wild type cDNAs could be expressed in vitro with similar efficiency. The three mutant enzymes thus synthesized showed catalytic activities comparable to that of the wild type oxidase. It is concluded that Tyr-55, Met-110 and His-217 are not directly involved in the catalytic function.
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D002384:Catalysis; D003605:D-Amino-Acid Oxidase / Q000235:genetics / Q000378:metabolism*; D004247:DNA / Q000235:genetics; D004274:DNA, Recombinant; D006639:Histidine; D007668:Kidney / Q000201:enzymology*; D008715:Methionine; D008969:Molecular Sequence Data; D009154:Mutation; D013329:Structure-Activity Relationship; D013552:Swine; D014443:Tyrosine
chemical_list: D004274:DNA, Recombinant; D014443:Tyrosine; D006639:Histidine; D004247:DNA; D008715:Methionine; D003605:D-Amino-Acid Oxidase