@article{e87e4124c6b644f0a5ba3e05f12be45f,
title = "Inhibitor Mimetic Mutations in the Pseudomonas aeruginosa PqsE Enzyme Reveal a Protein-Protein Interaction with the Quorum-Sensing Receptor RhlR That Is Vital for Virulence Factor Production",
abstract = "Pseudomonas aeruginosa is an opportunistic human pathogen that causes fatal infections. There exists an urgent need for new antimicrobial agents to combat P. aeruginosa. We conducted a screen for molecules that bind the virulence-controlling protein PqsE and characterized hit compounds for inhibition of PqsE enzymatic activity. The binding conformations of two inhibitory molecules, BB391 and BB393, were identified by crystallography, and inhibitor binding was mimicked by the substitution of PqsE residues E182 and S285 with tryptophan. Comparison of the inhibitor-mimetic mutations to the catalytically inactive PqsE D73A protein demonstrated that catalysis is not responsible for the role PqsE plays in driving virulence factor production. Rather, the PqsE E182W protein fails to interact with the quorum-sensing receptor, RhlR, and our results suggest that it is this interaction that is responsible for promoting virulence factor production in P. aeruginosa. These findings provide a new route for drug discovery efforts targeting PqsE.",
author = "Taylor, {Isabelle R.} and Paczkowski, {Jon E.} and Jeffrey, {Philip D.} and Henke, {Brad R.} and Smith, {Chari D.} and Bassler, {Bonnie L.}",
note = "Funding Information: We thank members of the Bassler laboratory for helpful advice and discussions, especially G. Whitney for performing the qRT-PCR experiments. The high-throughput DSF screen and initial enzyme assays with potential hit molecules were performed at Bienta. Molecules used in this study were synthesized at WuXi AppTec. Protein crystallography was performed in the Macromolecular Crystallography Core Facility at Princeton University. The AMX beamline of the National Synchrotron Light Source II, a U.S. Department of Energy (DOE) Office of Science User Facility operated for the DOE Office of Science by Brookhaven National Laboratory was also used under Contract DE-SC0012704. This work was supported by the Howard Hughes Medical Institute, NIH grant 2R37GM065859, and National Science Foundation grant MCB-1713731 to B. Bassler, NIH grant F32GM134583 to I. Taylor, and New York Community Trust Foundation grant P19-000454 to J. Paczkowski. The content herein is solely the responsibility of the authors and does not represent the official views of the National Institutes of Health. Publisher Copyright: {\textcopyright} 2021 The Authors. Published by American Chemical Society.",
year = "2021",
month = apr,
day = "16",
doi = "10.1021/acschembio.1c00049",
language = "English (US)",
volume = "16",
pages = "740--752",
journal = "ACS Chemical Biology",
issn = "1554-8929",
publisher = "American Chemical Society",
number = "4",
}