Affiliation:
1. Aventis Pharma France, 93235 Romainville
2. Enveloppes Bactériennes et Antibiotiques, UMR 8619 CNRS, Université de Paris-Sud, 91405 Orsay, France
Abstract
ABSTRACT
We have developed a novel assay specific to MraY, which catalyzes the first membrane step in the biosynthesis of bacterial cell wall peptidoglycan. This was accomplished by using UDP-MurNAc-
N
ε
-dansylpentapeptide, a fluorescent derivative of the MraY nucleotide substrate, and a partially purified preparation of MraY solubilized from membranes of an
Escherichia coli
overproducing strain. Two versions of the assay were developed, one consisting of the high-pressure liquid chromatography separation of the substrate and product (dansylated lipid I) and the other, without separation and adapted to the high-throughput format, taking advantage of the different fluorescence properties of the nucleotide and lipid I in the reaction medium. The latter assay was validated with a set of natural and synthetic MraY inhibitors.
Publisher
American Society for Microbiology
Subject
Infectious Diseases,Pharmacology (medical),Pharmacology
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5. mraY
Is an Essential Gene for Cell Growth in
Escherichia coli
Cited by
66 articles.
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