Identification of a S. aureus virulence factor by activity-based protein profiling (ABPP)

Abstract

Serine hydrolases play diverse roles in regulating host–pathogen interactions in a number of organisms, yet few have been characterized in the human pathogen Staphylococcus aureus. Here we describe a chemical proteomic screen that identified ten previously uncharacterized S. aureus serine hydrolases that mostly lack human homologs. We termed these enzymes fluorophosphonate-binding hydrolases (FphA–J). One hydrolase, FphB, can process short fatty acid esters, exhibits increased activity in response to host cell factors, is located predominantly on the bacterial cell surface in a subset of cells, and is concentrated in the division septum. Genetic disruption of fphB confirmed that the enzyme is dispensable for bacterial growth in culture but crucial for establishing infection in distinct sites in vivo. A selective small molecule inhibitor of FphB effectively reduced infectivity in vivo, suggesting that it may be a viable therapeutic target for the treatment or management of Staphylococcus infections.

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Fig. 1: Identification of serine hydrolases and inhibitors in live S. aureus by competitive ABPP.
Fig. 2: LC/LC-MS/MS-based identification of serine hydrolases in S. aureus.
Fig. 3: Biochemical characterization of FphB and development of an FphB-selective fluorescent ABP.
Fig. 4: Stimulatory activity of eukaryotic cell components on FphB activity.
Fig. 5: Imaging of FphB-activity using the fluorescent ABP JCP251-bT.
Fig. 6: Effects of loss of FphB activity on infectivity in vivo.

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Acknowledgements

We thank A. Horswill (University of Iowa) for sharing the GFP plasmid pCM29. We thank N. Amara and J. Yim for help with NMR analyses and S. Chen for LC-MS analysis of JCP678, and L. Popov, O. Zurek, J. Romaniuk and L. Cegelski for discussions. We also thank E. Yeh for access to the BD Accuri flow cytometer. C.S.L. was supported through a postdoctoral research fellowship by the German Research Foundation (DFG). This work was further supported through NIH grants 1R01GM111703 and R01EB026332 to M.B., 1R01GM117004 and 1R01GM118431-01A1 to E.W., 1R01AI101171 and 1R01AI069233 to E.P.S., and R21AI117255 to M.R.A.

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C.S.L. and M.B. conceived the project. C.S.L designed and performed the in vitro experiments, synthesized compounds and analyzed data. J.R.S. designed and performed the in vivo infection experiments and the genetic manipulation of S. aureus, and analyzed data. L.A.C. and E.W. performed LC-MS/MS analysis. R.C. contributed to the comparative bacterial labeling experiments. M.G. synthesized compounds. M.R.A. contributed to the experimental design and analyzed data. E.P.S. designed and analyzed in vivo infection experiments. M.B. supervised the project, designed experiments and analyzed data. C.S.L. and M.B. wrote the manuscript, and all authors reviewed, discussed and edited the manuscript.

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Correspondence to Matthew Bogyo.

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Lentz, C.S., Sheldon, J.R., Crawford, L.A. et al. Identification of a S. aureus virulence factor by activity-based protein profiling (ABPP). Nat Chem Biol 14, 609–617 (2018). https://doi.org/10.1038/s41589-018-0060-1

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