A High-Throughput, Homogeneous, Fluorescence Resonance Energy Transfer-Based Assay for Phospho-N-acetylmuramoyl-pentapeptide Translocase (MraY)
A High-Throughput, Homogeneous, Fluorescence Resonance Energy Transfer-Based Assay for Phospho-N-acetylmuramoyl-pentapeptide Translocase (MraY)
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DOI：
10.1177/1087057112436885
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发表时间：
2012-06-01
影响因子：
--
通讯作者：
Rivin, Olga
中科院分区：
文献类型：
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作者：
Shapiro, Adam B.;Jahic, Haris;Rivin, Olga
肽聚糖生物合成是细菌中的一个重要过程，因此是发现新的抗菌药物的合适靶点。肽聚糖生物合成的最后细胞质步骤之一由整合膜蛋白MraY催化，其将可溶性UDP-N-乙酰胞壁酰-五肽连接至膜结合受体磷酸十一异戊二烯酯。尽管已知几种天然产物衍生的MraY抑制剂，但没有一种具有作为抗菌药物临床使用所必需的性质。在这里，我们描述了一种新的，均匀的，基于荧光共振能量转移的MraY检测，适用于高通量筛选新的MraY抑制剂。该测定允许连续测量，或者可以在测量前淬灭。
Peptidoglycan biosynthesis is an essential process in bacteria and is therefore a suitable target for the discovery of new antibacterial drugs. One of the last cytoplasmic steps of peptidoglycan biosynthesis is catalyzed by the integral membrane protein MraY, which attaches soluble UDP-N-acetylmuramoyl-pentapeptide to the membrane-bound acceptor undecaprenyl phosphate. Although several natural product-derived inhibitors of MraY are known, none have the properties necessary to be of clinical use as antibacterial drugs. Here we describe a novel, homogeneous, fluorescence resonance energy transfer-based MraY assay that is suitable for high-throughput screening for novel MraY inhibitors. The assay allows for continuous measurement, or it can be quenched prior to measurement.