Fluorescence detection-based functional assay for high-throughput screening for MraY.

@article{Stachyra2004FluorescenceDF,
  title={Fluorescence detection-based functional assay for high-throughput screening for MraY.},
  author={Th{\'e}r{\`e}se Stachyra and Christophe Dini and Paul L Ferrari and Ahmed Bouhss and Jean van Heijenoort and Dominique Mengin-Lecreulx and Didier Blanot and Jacques Biton and Dominique Le Beller},
  journal={Antimicrobial agents and chemotherapy},
  year={2004},
  volume={48 3},
  pages={
          897-902
        }
}
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… CONTINUE READING

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