Development of a high-throughput fluorescence polarization assay for the discovery of phosphopantetheinyl transferase inhibitors.
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.
Duckworth
Benjamin P
BP
Center for Drug Design, University of Minnesota, Minneapolis, MN 55455, USA.
Aldrich
Courtney C
CC
D001419:Bacteria / Q000201:enzymology*; D001426:Bacterial Proteins / Q000037:antagonists & inhibitors* / Q000378:metabolism*; D055808:Drug Discovery; D004791:Enzyme Inhibitors / Q000494:pharmacology*; D005454:Fluorescence Polarization / Q000379:methods*; D057166:High-Throughput Screening Assays / Q000379:methods*; D008024:Ligands; D017855:Transferases (Other Substituted Phosphate Groups) / Q000037:antagonists & inhibitors* / Q000378:metabolism*
D001426:Bacterial Proteins; D004791:Enzyme Inhibitors; D008024:Ligands; C082662:phosphopantetheinyl transferase; D017855:Transferases (Other Substituted Phosphate Groups)