Amino acid divergence in the ligand-binding pocket of Vibrio LuxR/HapR proteins determines the efficacy of thiophenesulfonamide inhibitors
Abstract
The quorum-sensing signaling systems in Vibrio bacteria converge to control levels of the master transcription factors LuxR/HapR, a family of highly conserved proteins that regulate gene expression for bacterial behaviors. A compound library screen identified 2-thiophenesulfonamide compounds that specifically inhibit Vibrio campbellii LuxR but do not affect cell growth. We synthesized a panel of 50 thiophenesulfonamide compounds to examine the structure–activity relationship effects on Vibrio quorum sensing. The most potent molecule identified, PTSP (3-phenyl-1-(thiophen-2-ylsulfonyl)-1H-pyrazole), inhibits quorum sensing in multiple strains of V. vulnificus, V. parahaemolyticus, and V. campbellii at nanomolar concentrations. However, thiophenesulfonamide inhibition efficacy varies significantly among Vibrio species: PTSP is most inhibitory against V. vulnificus SmcR, but V. cholerae HapR is completely resistant to all thiophenesulfonamides tested. Reverse genetics experiments show that PTSP efficacy is dictated by amino acid sequence in the putative ligand-binding pocket: F75Y and C170F SmcR substitutions are each sufficient to eliminate PTSP inhibition. Further, in silico modeling distinguished the most potent thiophenesulfonamides from less-effective derivatives. Our results revealed the previously unknown differences in LuxR/HapR proteins that control quorum sensing in Vibrio species and underscore the potential for developing thiophenesulfonamides as specific quorum sensing-directed treatments for Vibrio infections.
Additional Information
© 2021 John Wiley & Sons Ltd. Issue Online: 23 October 2021; Version of Record online: 14 September 2021; Accepted manuscript online: 01 September 2021; Manuscript accepted: 26 August 2021; Manuscript revised: 09 August 2021; Manuscript received: 30 April 2021. The authors thank Stanna Dorn and the Indiana University students in Organic Chemistry C344 laboratory classes for synthesis of molecules in the thiophenesulfonamide panel. They thank the Indiana University School of Medicine Chemical Genomics Core Facility for assistance with chemical screens and analyses. They thank Dr. Irene Newton for assistance with data analysis. They also also thank Dr. Giovanni Gonzalez-Gutierrez for assistance with ITC experiments and analysis. This work was supported by National Institutes of Health grant R35GM124698 to JVK and with support from the Indiana Clinical and Translational Sciences Institute funded, in part by Award Number UL1TR002529 from the National Institutes of Health, National Center for Advancing Translational Sciences, Clinical and Translational Sciences Award. The content is solely the responsibility of the authors and does not necessarily represent the official views of the National Institutes of Health. Author Contributions: JVK and LCB designed the experiments; JVK, JDN, PS, JC, ES, MEM, and LCB performed experiments; JVK, JDN, RH, JC, ES, MEM and LCB analyzed experimental results; and JVK, JDN, and LCB wrote the manuscript. The authors declare that they have no conflicts of interest. Data Availability Statement: The data that support the findings of this study are available from the corresponding author upon reasonable request.Attached Files
Accepted Version - nihms-1846872.pdf
Submitted - 2021.04.14.439868v1.full.pdf
Supplemental Material - mmi14804-sup-0001-supinfo.pdf
Supplemental Material - mmi14804-sup-0002-supinfo.xlsx
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Additional details
- Alternative title
- Thiophenesulfonamides are specific inhibitors of quorum sensing in pathogenic Vibrios
- PMCID
- PMC9682466
- Eprint ID
- 108737
- Resolver ID
- CaltechAUTHORS:20210415-074717081
- R35GM124698
- NIH
- Indiana Clinical and Translational Sciences Institute
- UL1TR002529
- NIH
- Created
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2021-04-16Created from EPrint's datestamp field
- Updated
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2023-07-07Created from EPrint's last_modified field