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

Benjamin P. Duckworth, Courtney C. Aldrich

Research output: Contribution to journalArticlepeer-review

21 Scopus citations

Abstract

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.

Original languageEnglish (US)
Pages (from-to)13-19
Number of pages7
JournalAnalytical Biochemistry
Volume403
Issue number1-2
DOIs
StatePublished - Aug 2010

Keywords

  • Enzyme assay
  • Fluorescence polarization
  • High-throughput screening
  • Phosphopantetheinyl transferase
  • Sfp

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