Open AccessThe role of histidine residues conserved in the putative ATP‐binding region of macrolide 2′‐phosphotransferase II
Author(s) -
Taniguchi Kazuo,
Nakamura Akio,
Tsurubuchi Kazue,
O'Hara Koji,
Sawai Tetsuo
Publication year - 2004
Publication title -
fems microbiology letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.899
H-Index - 151
eISSN - 1574-6968
pISSN - 0378-1097
DOI - 10.1016/s0378-1097(03)00961-3
Subject(s) - oleandomycin , phosphotransferase , enzyme , biochemistry , histidine , conserved sequence , mutant , binding site , chemistry , biology , stereochemistry , peptide sequence , antibiotics , erythromycin , gene
Macrolide 2′‐phosphotransferase (MPH(2′)) catalyzes the transfer of the γ‐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.