Studying aminoglycoside modification by the acetyltransferase class of resistance-causing enzymes via microarray

Aminoglycoside microarrays have been used to study modification of sugars by the acetyltransferase class of resistance enzymes. Aminoglycosides are broad-spectrum antibacterials to which some bacteria have acquired resistance. The most common mode of resistance to aminoglycosides is enzymatic modifi...

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Bibliographic Details
Published in:Carbohydrate research 2008-11, Vol.343 (17), p.2924-2931
Main Authors: Barrett, Olivia J., Pushechnikov, Alexei, Wu, Meilan, Disney, Matthew D.
Format: Article
Language:English
Subjects:
Acetyltransferases - metabolism
Aminoglycosides
Aminoglycosides - chemistry
Aminoglycosides - pharmacology
Anti-Bacterial Agents - pharmacology
Antibiotic resistance
Bacterial Proteins - drug effects
Bacterial Proteins - metabolism
Carbohydrate Conformation
Carbohydrate modification
Drug Resistance, Bacterial
Escherichia coli - enzymology
Microarrays
Models, Molecular
Mycobacterium tuberculosis - enzymology
Oligonucleotide Array Sequence Analysis - methods
RNA, Bacterial - drug effects
RNA, Bacterial - metabolism
RNA, Ribosomal - drug effects
RNA, Ribosomal - metabolism
Substrate Specificity
Tobramycin - pharmacology
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Summary:Aminoglycoside microarrays have been used to study modification of sugars by the acetyltransferase class of resistance enzymes. 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.
ISSN:0008-6215
1873-426X
DOI:10.1016/j.carres.2008.08.018