Chemoproteomic profiling of host and pathogen enzymes active in cholera

An Erratum to this article was published on 18 May 2016

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Abstract

Activity-based protein profiling (ABPP) is a chemoproteomic tool for detecting active enzymes in complex biological systems. We used ABPP to identify secreted bacterial and host serine hydrolases that are active in animals infected with the cholera pathogen Vibrio cholerae. Four V. cholerae proteases were consistently active in infected rabbits, and one, VC0157 (renamed IvaP), was also active in human choleric stool. Inactivation of IvaP influenced the activity of other secreted V. cholerae and rabbit enzymes in vivo, and genetic disruption of all four proteases increased the abundance of intelectin, an intestinal lectin, and its binding to V. cholerae in infected rabbits. Intelectin also bound to other enteric bacterial pathogens, suggesting that it may constitute a previously unrecognized mechanism of bacterial surveillance in the intestine that is inhibited by pathogen-secreted proteases. Our work demonstrates the power of activity-based proteomics to reveal host-pathogen enzymatic dialog in an animal model of infection.

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Figure 1: ABPP detects active serine hydrolases in rabbit cecal fluid and human choleric stool.
Figure 2: Predicted protein domain structures of IvaP, VCA0812, VesA and VesB.
Figure 3: IvaP undergoes growth phase–dependent processing and autoproteolysis.
Figure 4: V. cholerae proteases alter the serine hydrolase activity, lipid profile and protein content of rabbit cecal fluid.
Figure 5: V. cholerae proteases decrease intelectin binding to V. cholerae in infected rabbits.

Change history

  • 23 March 2016

    Within the Discussion section, one instance referring to the published crystal structure of trimeric human intelectin-1 was attributed to reference 47 instead of the correct reference 44. The error has been corrected in the HTML and PDF versions of the article.

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Acknowledgements

We gratefully acknowledge R. LaRocque, Y. Millet and J. Lee for reagents and technical assistance, and members of M.K.W.'s lab for helpful discussions. We also thank V. Carey for the statistical analysis of MS data and D. Bak for help formatting MS data sets. This work was supported by the National Institutes of Health (R37 AI-042347 to M.K.W., F31 AI-120665 to T.H., R01 AI-106878 and U01 AI-058935 to E.T.R., Institutional Training Grant T32 DK 7477-30 to S.K.H.), the Howard Hughes Medical Institute (M.K.W.), the Charles A. King Trust Postdoctoral Fellowship Program, Bank of America, N.A., Co-Trustee (S.K.H.), the Damon Runyon Cancer Research Foundation (DRR-18-12 to E.W.), the Smith Family Foundation (E.W.) and the Swiss National Science Foundation (P300P3_155287/1 to S.A.).

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Authors

Contributions

S.K.H., E.W. and M.K.W. conceived the project and designed the experiments. S.K.H. generated the V. cholerae mutant strains, performed all biochemistry and ABPP experiments and analyzed the MS data. S.A., T.H. and D.M. performed the rabbit infections. J.M. performed the MS experiments and assisted with MS data analysis. J.S. provided technical guidance on intelectin assays. L.C. assisted with the construction of V. cholerae mutant strains. D.A.B. synthesized FP-biotin. F.Q. and E.T.R. provided the human choleric stool used in this study. S.K.H., B.M.D., and M.K.W. wrote the manuscript.

Corresponding authors

Correspondence to Eranthie Weerapana or Matthew K Waldor.

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The authors declare no competing financial interests.

Supplementary information

Supplementary Text and Figures

Supplementary Results, Supplementary Tables 1–9 and Supplementary Figures 1–10. (PDF 7161 kb)

Supplementary Data Set 1

FP-biotin-enriched proteins detected in wild-type cecal fluid, comparative ABPP-MudPIT analysis of wild-type and S361A cecal fluid and combined unfiltered data from all ABPP-MudPIT analyses of wild-type cecal fluid. (XLSX 92 kb)

Supplementary Data Set 2

FP-biotin–enriched proteins from human choleric stool supernatant and unfiltered data from ABPP-MudPIT analysis of human choleric stool supernatant. (XLSX 28 kb)

Supplementary Data Set 3

ABPP-MudPIT analysis of wild-type and ΔivaP biofilm culture supernatants. (XLSX 23 kb)

Supplementary Data Set 4

ABPP-MudPIT analysis of exponential- and stationary-phase V. cholerae cell lysates. (XLSX 40 kb)

Supplementary Data Set 5

Comparative total and free fatty acid analysis of wild-type and Δquad cecal fluid. (XLSX 21 kb)

Supplementary Data Set 6

V. cholerae and rabbit proteins detected in unfractionated wild-type and Δquad cecal fluid. (XLSX 188 kb)

Supplementary Data Set 7

V. cholerae and human proteins detected in unfractionated human choleric stool. (XLSX 52 kb)

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Hatzios, S., Abel, S., Martell, J. et al. Chemoproteomic profiling of host and pathogen enzymes active in cholera. Nat Chem Biol 12, 268–274 (2016). https://doi.org/10.1038/nchembio.2025

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