title: Studying aminoglycoside modification by the acetyltransferase class of resistance-causing enzymes via microarray.
abstract: Aminoglycosides are broad-spectrum antibacterials to which some bacteria have acquired resistance. The most common mode of resistance to aminoglycosides is enzymatic modification of the drug by different classes of enzymes including acetyltransferases (AACs). Thus, the modification of aminoglycosides by AAC(2') from Mycobacterium tuberculosis and AAC(3) from Escherichia coli was studied using aminoglycoside microarrays. Results show that both enzymes modify their substrates displayed on an array surface in a manner that mimics their relative levels of modification in solution. Because aminoglycosides that are modified by resistance-causing enzymes have reduced affinities for binding their therapeutic target, the bacterial rRNA aminoacyl-tRNA site (A-site), arrays were probed for binding to a fluorescently labeled oligonucleotide mimic of the A-site after modification. A decrease in binding was observed when aminoglycosides were modified by AAC(3). In contrast, a decrease in binding of the A-site is not observed when aminoglycosides are modified by AAC(2'). Interestingly, these effects mirror the biological functions of the enzymes: the AAC(3) used in this study is known to confer aminoglycoside resistance, while the AAC(2') is chromosomally encoded and unlikely to play a role in resistance. These studies lay a direct foundation for studying resistance to aminoglycosides and can also have more broad applications in identifying and studying non-aminoglycoside carbohydrates or proteins as substrates for acetyltransferase enzymes.
authors.affiliation: Department of Chemistry, The Center for Excellence in Bioinformatics and Life Sciences, University at Buffalo, 657 Natural Sciences Complex, Buffalo, NY 14260, USA.
authors.forename: Olivia J
authors.initials: OJ
authors.lastname: Barrett
authors.forename: Alexei
authors.initials: A
authors.lastname: Pushechnikov
authors.forename: Meilan
authors.initials: M
authors.lastname: Wu
authors.forename: Matthew D
authors.initials: MD
authors.lastname: Disney
mesh_terms: D000123:Acetyltransferases / Q000378:metabolism*; D000617:Aminoglycosides / Q000737:chemistry / Q000494:pharmacology*; D000900:Anti-Bacterial Agents / Q000494:pharmacology; D001426:Bacterial Proteins / Q000187:drug effects / Q000378:metabolism; D002236:Carbohydrate Conformation; D024881:Drug Resistance, Bacterial; D004926:Escherichia coli / Q000201:enzymology; D008958:Models, Molecular; D009169:Mycobacterium tuberculosis / Q000201:enzymology; D020411:Oligonucleotide Array Sequence Analysis / Q000379:methods*; D012329:RNA, Bacterial / Q000187:drug effects / Q000378:metabolism; D012335:RNA, Ribosomal / Q000187:drug effects / Q000378:metabolism; D013379:Substrate Specificity; D014031:Tobramycin / Q000494:pharmacology
chemical_list: D000617:Aminoglycosides; D000900:Anti-Bacterial Agents; D001426:Bacterial Proteins; D012329:RNA, Bacterial; D012335:RNA, Ribosomal; D000123:Acetyltransferases; D014031:Tobramycin
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