The identification of the aminoglycoside-phosphorylating enzymes APH(2'') and APH(3') from the characterization of their reaction products by high performance liquid chromatography.
A method is described for the identification of the aminoglycoside-phosphorylating (APH) enzymes APH(2'') and APH(3') by the high performance liquid chromatographic (HPLC) identification of their products of reaction with kanamycin and ATP. Twelve reference strains which produce either APH(2'') or APH(3') were examined by both this method and the radioenzymatic profile method; correct enzyme identification was made with 12/12 of the strains by the HPLC method and 11/12 of the strains by the radioenzymatic profile method. Reaction products of APH(2'') or APH(3') with ATP and butirosin, geneticin, lividomycin, ribostamycin, sissomicin or tobramycin were also characterized by HPLC. Conditions for their chromatography are given.
D000617:Aminoglycosides / Q000378:metabolism / Q000494:pharmacology; D000900:Anti-Bacterial Agents / Q000378:metabolism* / Q000494:pharmacology; D001419:Bacteria / Q000187:drug effects / Q000201:enzymology*; D002851:Chromatography, High Pressure Liquid; D004352:Drug Resistance, Microbial; D007612:Kanamycin / Q000378:metabolism; D019868:Kanamycin Kinase; D010766:Phosphorylation; D010770:Phosphotransferases / Q000032:analysis*; D017853:Phosphotransferases (Alcohol Group Acceptor); D009764:o-Phthalaldehyde / Q000032:analysis
D000617:Aminoglycosides; D000900:Anti-Bacterial Agents; D007612:Kanamycin; D009764:o-Phthalaldehyde; D010770:Phosphotransferases; D017853:Phosphotransferases (Alcohol Group Acceptor); C048564:aminoglycoside 2''-phosphotransferase; D019868:Kanamycin Kinase