PMID:14982781
| Citation |
Stachyra, T, Dini, C, Ferrari, P, Bouhss, A, van Heijenoort, J, Mengin-Lecreulx, D, Blanot, D, Biton, J and Le Beller, D (2004) Fluorescence detection-based functional assay for high-throughput screening for MraY. Antimicrob. Agents Chemother. 48:897-902 |
|---|---|
| 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(epsilon)-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. |
| Links | |
| Keywords |
Anti-Bacterial Agents/pharmacology; Bacteria/drug effects; Bacteria/genetics; Bacterial Proteins/antagonists & inhibitors; Chromatography, High Pressure Liquid; Dansyl Compounds/chemistry; Dansyl Compounds/pharmacology; Drug Evaluation, Preclinical; Escherichia coli/drug effects; Escherichia coli/enzymology; Microbial Sensitivity Tests; Monosaccharides/metabolism; Oligopeptides/metabolism; Oligopeptides/pharmacology; Plasmids/drug effects; Plasmids/genetics; Spectrometry, Fluorescence; Spectrophotometry, Ultraviolet; Transferases/antagonists & inhibitors |
| edit table |
Significance
You can help EcoliWiki by summarizing why this paper is useful
Useful Materials and Methods
You can help Ecoliwiki by describing the useful materials (strains, plasmids, antibodies, etc) described in this paper.
Annotations
<annotationlinks/>
EcoliWiki Links
Add links to pages that link here (e.g. gene, product, method pages)
References
See Help:References for how to manage references in EcoliWiki.