Development of a high-throughput fluorescence polarization assay for the discovery of phosphopantetheinyl transferase inhibitors

被引：29

作者：

Duckworth, Benjamin P. ^{[1
]}

Aldrich, Courtney C. ^{[1
]}

机构：

[1] Univ Minnesota, Ctr Drug Design, Minneapolis, MN 55455 USA

来源：

ANALYTICAL BIOCHEMISTRY | 2010年 / 403卷 / 1-2期

关键词：

Sfp; Phosphopantetheinyl transferase; Fluorescence polarization; High-throughput screening; Enzyme assay; MODULAR PEPTIDE SYNTHETASES; PROTEIN-COUPLED RECEPTORS; SMALL-MOLECULE INHIBITORS; BACILLUS-SUBTILIS; SIDEROPHORE BIOSYNTHESIS; SCREENING ASSAYS; SFP; POLYKETIDE; MECHANISM; SUPERFAMILY;

D O I：

10.1016/j.ab.2010.04.009

中图分类号：

Q5 [生物化学];

学科分类号：

071010 ; 081704 ;

摘要：

An alarming number of clinically relevant bacterial pathogens are becoming resistant to many antibiotics, thereby fueling intense research into the discovery of novel therapeutic targets. Phosphopantetheinyl transferases (PPTases) represent a promising target for antibacterial development because these enzymes are crucial for the biosynthesis of a multitude of a pathogen's collection of essential metabolites and virulence factors biosynthesized via polyketide synthase (PKS) and nonribosomal peptide synthetase (NRPS) pathways. Here we describe the development of a fluorescence polarization (FP) assay that is amenable for high-throughput screening to identify PPTase inhibitors. The FP assay was validated against a panel of competitive ligands and displayed an excellent Z' score. (C) 2010 Elsevier Inc. All rights reserved.

引用

页码：13 / 19

页数：7

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