A MurG assay which utilises a synthetic analogue of lipid I
- 1 February 2003
- journal article
- research article
- Published by Oxford University Press (OUP) in FEMS Microbiology Letters
- Vol. 219 (1) , 115-119
- https://doi.org/10.1016/s0378-1097(02)01203-x
Abstract
A standard assay for the MurG enzyme using a lipid I analogue [MurNAc( N ɛ-dansylpentapeptide)-pyrophosphoryl ( R , S )-α-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.Keywords
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