title: Identification of functional amino acids in the macrolide 2'-phosphotransferase II.
abstract: Macrolide 2'-phosphotransferase [MPH(2')] transfers the gamma phosphate of ATP to the 2'-OH group of macrolide antibiotics. The role of aspartic acids in the putative ATP-binding site of MPH(2')II was investigated through the substitution of alanine for aspartate by site-directed mutagenesis. D200A, D209A, D219A, and D231A mutant strains were unable to inactivate the substrate oleandomycin, while a D227A mutant retained 7% of the activity of the original enzyme.
authors.affiliation: Division of Microbial Chemistry, Faculty of Pharmaceutical Sciences, Chiba University, Inage-ku, Chiba 263-8522, Japan.
authors.forename: K
authors.initials: K
authors.lastname: Taniguchi
authors.forename: A
authors.initials: A
authors.lastname: Nakamura
authors.forename: K
authors.initials: K
authors.lastname: Tsurubuchi
authors.forename: A
authors.initials: A
authors.lastname: Ishii
authors.forename: K
authors.initials: K
authors.lastname: O'Hara
authors.forename: T
authors.initials: T
authors.lastname: Sawai
mesh_terms: D000255:Adenosine Triphosphate / Q000378:metabolism; D000409:Alanine / Q000235:genetics / Q000378:metabolism*; D000595:Amino Acid Sequence; D000900:Anti-Bacterial Agents / Q000493:pharmacokinetics / Q000494:pharmacology; D001224:Aspartic Acid / Q000235:genetics / Q000378:metabolism*; D001665:Binding Sites; D004352:Drug Resistance, Microbial; D004926:Escherichia coli / Q000187:drug effects / Q000201:enzymology / Q000235:genetics; D029968:Escherichia coli Proteins; D008969:Molecular Sequence Data; D016297:Mutagenesis, Site-Directed; D009827:Oleandomycin / Q000493:pharmacokinetics / Q000494:pharmacology; D017853:Phosphotransferases (Alcohol Group Acceptor) / Q000235:genetics / Q000378:metabolism*; D017386:Sequence Homology, Amino Acid; D013379:Substrate Specificity
chemical_list: D000900:Anti-Bacterial Agents; D029968:Escherichia coli Proteins; D001224:Aspartic Acid; D000255:Adenosine Triphosphate; D017853:Phosphotransferases (Alcohol Group Acceptor); C061369:macrolide 2'-kinase; C501557:mphB protein, E coli; D000409:Alanine; D009827:Oleandomycin