A Time‐Resolved FRET Assay Identifies a Small Molecule that Inhibits the Essential Bacterial Cell Wall Polymerase FtsW

• Published in: Angewandte Chemie International Edition Vol. 62; no. 25; pp. e202301522 - n/a
• Main Authors: Park, Youngseon, Taguchi, Atsushi, Baidin, Vadim, Kahne, Daniel, Walker, Suzanne
• Format: Journal Article
• Language: English
• Published: Germany Wiley Subscription Services, Inc 19-06-2023
• Edition: International ed. in English
• Subjects: Antibiotics; Assay Development; Assaying; Bacteria; Bacterial Proteins - metabolism; Cell division; Cell survival; Cell Wall - metabolism; Cell walls; Elongation; Energy transfer; Fluorescence Resonance Energy Transfer; Glycan; High-Throughput Screening; Inhibitors; Lipids; Membrane Proteins - metabolism; Penicillin-Binding Proteins - metabolism; Peptidoglycan; Peptidoglycan - metabolism; Peptidoglycan Glycosyltransferase - metabolism; Peptidoglycans; Polymerization; Proteins; Sporulation; Staphylococcus aureus - metabolism; Strands; High-Throughput Screening; Peptidoglycan; Bacteria; Antibiotics; Assay Development
• ISSN: 1433-7851; 1521-3773; 1521-3773
• DOI: 10.1002/anie.202301522

The peptidoglycan cell wall is essential for bacterial survival. To form the cell wall, peptidoglycan glycosyltransferases (PGTs) polymerize Lipid II to make glycan strands and then those strands are crosslinked by transpeptidases (TPs). Recently, the SEDS (for shape, elongation, division, and sporulation) proteins were identified as a new class of PGTs. The SEDS protein FtsW, which produces septal peptidoglycan during cell division, is an attractive target for novel antibiotics because it is essential in virtually all bacteria. Here, we developed a time‐resolved Förster resonance energy transfer (TR‐FRET) assay to monitor PGT activity and screened a Staphylococcus aureus lethal compound library for FtsW inhibitors. We identified a compound that inhibits S. aureus FtsW in vitro. Using a non‐polymerizable Lipid II derivative, we showed that this compound competes with Lipid II for binding to FtsW. The assays described here will be useful for discovering and characterizing other PGT inhibitors.
FtsW is a newly discovered peptidoglycan polymerase essential for cell division. We developed a time‐resolved Förster resonance energy transfer (TR‐FRET) assay, which we used to screen a Staphylococcus aureus lethal compound library for FtsW inhibitors. We identified a small molecule that inhibits FtsW in vitro. Using a fluorescently labeled, non‐polymerizable Lipid II derivative, we showed that this compound displaces Lipid II from FtsW.