title: The role of histidine residues conserved in the putative ATP-binding region of macrolide 2'-phosphotransferase II.
abstract: Macrolide 2'-phosphotransferase (MPH(2')) catalyzes the transfer of the gamma-phosphate of ATP to the 2'-hydroxyl group of macrolide antibiotics. In this study, H198 and H205, conserved in the ATP-binding region motif 1 in the putative amino acid sequence of MPH(2')II, were replaced by Ala to investigate their role. H205 was also subsequently replaced by Asn. H198A and H205N mutant enzymes retained more than 50% of the specific activity of the original enzyme to substrate oleandomycin. On the other hand, the specific activity of the H205A mutant enzyme was reduced to less than 1% of that of the wild enzyme. The results suggested that H205 is crucial for maintaining the catalytic activity of MPH(2')II, and Asn can substitute for His at this position.
authors.affiliation: Division of Microbial Chemistry, Faculty of Pharmaceutical Sciences, Chiba University, 1-33, Yayoi-cho, Inage-ku, Chiba 263-8522, Japan.
authors.forename: Kazuo
authors.initials: K
authors.lastname: Taniguchi
authors.forename: Akio
authors.initials: A
authors.lastname: Nakamura
authors.forename: Kazue
authors.initials: K
authors.lastname: Tsurubuchi
authors.forename: Koji
authors.initials: K
authors.lastname: O'Hara
authors.forename: Tetsuo
authors.initials: T
authors.lastname: Sawai
mesh_terms: D000255:Adenosine Triphosphate / Q000378:metabolism*; D000595:Amino Acid Sequence; D000900:Anti-Bacterial Agents / Q000494:pharmacology; D017124:Conserved Sequence; D024881:Drug Resistance, Bacterial / Q000235:genetics; D004926:Escherichia coli / Q000187:drug effects / Q000235:genetics*; D029968:Escherichia coli Proteins; D006639:Histidine / Q000235:genetics; D017853:Phosphotransferases (Alcohol Group Acceptor) / Q000235:genetics* / Q000378:metabolism*; D015572:Spiramycin / Q000494:pharmacology; D015645:Tylosin / Q000494:pharmacology
chemical_list: D000900:Anti-Bacterial Agents; D029968:Escherichia coli Proteins; D006639:Histidine; D015572:Spiramycin; D000255:Adenosine Triphosphate; D017853:Phosphotransferases (Alcohol Group Acceptor); C061369:macrolide 2'-kinase; C501557:mphB protein, E coli; D015645:Tylosin