Inhibition of Fosfomycin Resistance Protein FosB from Gram-Positive Pathogens by Phosphonoformate

Skye Travis; Keith D. Green; Nathaniel C. Gilbert; Oleg Tsodikov; Sylvie Garneau-Tsodikova; Matthew K. Thompson

doi:10.1021/acs.biochem.2c00566

Inhibition of Fosfomycin Resistance Protein FosB from Gram-Positive Pathogens by Phosphonoformate

Skye Travis, Keith D. Green, Nathaniel C. Gilbert, Oleg Tsodikov, Sylvie Garneau-Tsodikova, Matthew K. Thompson

Research output: Contribution to journal › Article › peer-review

2 Scopus citations

Abstract

The Gram-positive pathogen Staphylococcus aureus is a leading cause of antimicrobial resistance related deaths worldwide. Like many pathogens with multidrug-resistant strains, S. aureus contains enzymes that confer resistance through antibiotic modification(s). One such enzyme present in S. aureus is FosB, a Mn²⁺-dependent l-cysteine or bacillithiol (BSH) transferase that inactivates the antibiotic fosfomycin. fosB gene knockout experiments show that the minimum inhibitory concentration (MIC) of fosfomycin is significantly reduced when the FosB enzyme is not present. This suggests that inhibition of FosB could be an effective method to restore fosfomycin activity. We used high-throughput in silico-based screening to identify small-molecule analogues of fosfomycin that inhibited thiol transferase activity. Phosphonoformate (PPF) was a top hit from our approach. Herein, we have characterized PPF as a competitive inhibitor of FosB from S. aureus (FosB^Sa) and Bacillus cereus (FosB^Bc). In addition, we have determined a crystal structure of FosB^Bc with PPF bound in the active site. Our results will be useful for future structure-based development of FosB inhibitors that can be delivered in combination with fosfomycin in order to increase the efficacy of this antibiotic.

Original language	English
Pages (from-to)	109-117
Number of pages	9
Journal	Biochemistry
Volume	62
Issue number	1
DOIs	https://doi.org/10.1021/acs.biochem.2c00566
State	Published - Jan 3 2023

Bibliographical note

Funding Information:
This work was funded by The University of Alabama startup funds (to M.K.T.) and the Cystic Fibrosis Foundation Grant #THOMPS2010 (to M.K.T.). S.T. would like to acknowledge the Department of Education GAANN Grant #P200A150329. M.K.T would like to acknowledge the generosity of the Richard N. Armstrong family and Vanderbilt University Department of Biochemistry for initial laboratory equipment along with plasmids and reagents specific to this project. This work is based upon research conducted at the Northeastern Collaborative Access Team beamlines, which are funded by the National Institute of General Medical Sciences from the National Institutes of Health (P30 GM124165). The Eiger 16M detector on the 24-ID-E beam line is funded by a NIH-ORIP HEI grant (S10OD021527). This research used resources of the Advanced Photon Source, a U.S. Department of Energy (DOE) Office of Science User Facility operated for the DOE Office of Science by Argonne National Laboratory under Contract No. DE- AC02-06CH11357.

Publisher Copyright:
© 2022 American Chemical Society.

ASJC Scopus subject areas

Biochemistry

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title = "Inhibition of Fosfomycin Resistance Protein FosB from Gram-Positive Pathogens by Phosphonoformate",

abstract = "The Gram-positive pathogen Staphylococcus aureus is a leading cause of antimicrobial resistance related deaths worldwide. Like many pathogens with multidrug-resistant strains, S. aureus contains enzymes that confer resistance through antibiotic modification(s). One such enzyme present in S. aureus is FosB, a Mn2+-dependent l-cysteine or bacillithiol (BSH) transferase that inactivates the antibiotic fosfomycin. fosB gene knockout experiments show that the minimum inhibitory concentration (MIC) of fosfomycin is significantly reduced when the FosB enzyme is not present. This suggests that inhibition of FosB could be an effective method to restore fosfomycin activity. We used high-throughput in silico-based screening to identify small-molecule analogues of fosfomycin that inhibited thiol transferase activity. Phosphonoformate (PPF) was a top hit from our approach. Herein, we have characterized PPF as a competitive inhibitor of FosB from S. aureus (FosBSa) and Bacillus cereus (FosBBc). In addition, we have determined a crystal structure of FosBBc with PPF bound in the active site. Our results will be useful for future structure-based development of FosB inhibitors that can be delivered in combination with fosfomycin in order to increase the efficacy of this antibiotic.",

author = "Skye Travis and Green, {Keith D.} and Gilbert, {Nathaniel C.} and Oleg Tsodikov and Sylvie Garneau-Tsodikova and Thompson, {Matthew K.}",

note = "Funding Information: This work was funded by The University of Alabama startup funds (to M.K.T.) and the Cystic Fibrosis Foundation Grant #THOMPS2010 (to M.K.T.). S.T. would like to acknowledge the Department of Education GAANN Grant #P200A150329. M.K.T would like to acknowledge the generosity of the Richard N. Armstrong family and Vanderbilt University Department of Biochemistry for initial laboratory equipment along with plasmids and reagents specific to this project. This work is based upon research conducted at the Northeastern Collaborative Access Team beamlines, which are funded by the National Institute of General Medical Sciences from the National Institutes of Health (P30 GM124165). The Eiger 16M detector on the 24-ID-E beam line is funded by a NIH-ORIP HEI grant (S10OD021527). This research used resources of the Advanced Photon Source, a U.S. Department of Energy (DOE) Office of Science User Facility operated for the DOE Office of Science by Argonne National Laboratory under Contract No. DE- AC02-06CH11357. Publisher Copyright: {\textcopyright} 2022 American Chemical Society.",

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AU - Thompson, Matthew K.

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Inhibition of Fosfomycin Resistance Protein FosB from Gram-Positive Pathogens by Phosphonoformate

Abstract

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