A MurG assay which utilises a synthetic analogue of lipid I

被引：32

作者：

Auger, G ^{[1
]}

van Heijenoort, J ^{[1
]}

Mengin-Lecreulx, D ^{[1
]}

Blanot, D ^{[1
]}

机构：

[1] Univ Paris 11, UMR 8619 CNRS, F-91405 Orsay, France

来源：

FEMS MICROBIOLOGY LETTERS | 2003年 / 219卷 / 01期

关键词：

MurG; lipid I; lipid II; peptidoglycan; dimethylsulfoxide;

D O I：

10.1016/S0378-1097(02)01203-X

中图分类号：

Q93 [微生物学];

学科分类号：

071005 ; 100705 ;

摘要：

A standard assay for the MurG enzyme using a lipid I analogue [MurNAc(N-epsilon-dansylpentapeptide)-pyrophosphoryl (R,S)-alpha-dihydroheptaprenol] and radioactive UDP-N-acetylglucosamine was set up. A high concentration (35%) of dimethylsulfoxide was necessary for maximal activity. Separation and quantitation were accomplished by reverse-phase high performance liquid chromatography (HPLC) in isocratic conditions and on-line radioactivity detection, thereby providing a rapid and accurate assay. The kinetic parameters of the MurG reaction were determined; the reaction was shown to also catalyse the reverse reaction at a measurable rate. A lipid I analogue containing dihydroundecaprenol as the prenyl chain turned out to be a poor MurG substrate, presumably owing to aggregation. (C) 2003 Federation of European Microbiological Societies. Published by Elsevier Science B.V. All rights reserved.

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页码：115 / 119

页数：5

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