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Colibactin assembly line enzymes use S-adenosylmethionine to build a cyclopropane ring

Nature Chemical Biology volume 13, pages 1063–1065 (2017)Cite this article

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Abstract

Despite containing an α-amino acid, the versatile cofactor S-adenosylmethionine (SAM) is not a known building block for nonribosomal peptide synthetase (NRPS) assembly lines. Here we report an unusual NRPS module from colibactin biosynthesis that uses SAM for amide bond formation and subsequent cyclopropanation. Our findings showcase a new use for SAM and reveal a novel biosynthetic route to a functional group that likely mediates colibactin's genotoxicity.

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Figure 1: SAM is activated as a nonribosomal peptide synthetase (NRPS) building block.
Figure 2: ClbH-mediated SAM elongation precedes ClbI-catalyzed cyclopropanation.

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Acknowledgements

We thank C. Brotherton and A. Sieg for help with cloning; J. May (Kahne lab, Harvard University, Cambridge, MA) for help with radiometric assays; M. McCallum for help with bioinformatic analyses; and P. Boudreau, C. Chittim, D. Kenny, H. Nakamura, and S. Peck for helpful discussions. L.Z., Y.J., M.R.W., and E.P.B. acknowledge financial support from National Cancer Institute (1R01CA208834-01), the Damon Runyon–Rachleff Innovation Award, and the Packard Fellowship for Science and Engineering. M.T.H. and N.L.K. were supported by Northwestern University and the US National Institutes of Health (GM 067725 and AT 009143). M.R.W. is supported by an American Cancer Society-New England Division Postdoctoral Fellowship (PF-16-122-01-CDD).

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Author notes

  1. Matthew T Henke

    Present address: Department of Biological Chemistry and Molecular Pharmacology, Harvard Medical School, Boston, Massachusetts, USA

  2. Yindi Jiang, Matthew T Henke and Matthew R Wilson: These authors contributed equally to this work.

Authors and Affiliations

  1. Department of Chemistry and Chemical Biology, Harvard University, Cambridge, Massachusetts, USA

    Li Zha, Yindi Jiang, Matthew R Wilson & Emily P Balskus

  2. Department of Chemistry, Northwestern University, Evanston, Illinois, USA

    Matthew T Henke & Neil L Kelleher

  3. FAS Division of Science, Small Molecule Mass Spectrometry Facility, Cambridge, Massachusetts, USA

    Jennifer X Wang

  4. Department of Molecular Biosciences and the Feinberg School of Medicine, Northwestern University, Evanston, Illinois, USA

    Neil L Kelleher

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  1. Li Zha
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Contributions

L.Z. cloned, overexpressed, and purified colibactin biosynthetic enzymes and completed all enzymatic assays and bioinformatic analyses. Y.J. generated mutants used in the study and prepared culture extracts. M.T.H. analyzed enzymatic assays by protein mass spectrometry. M.R.W. prepared and characterized synthetic standards. J.X.W. helped to develop the analysis method for small-molecule LC–MS/MS. All coauthors collected and analyzed data. L.Z., M.T.H., N.L.K., and E.P.B. prepared and revised the manuscript.

Corresponding authors

Correspondence to Neil L Kelleher or Emily P Balskus.

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The authors declare no competing financial interests.

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Supplementary Results, Supplementary Tables 1–6 and Supplementary Figures 1–26 (PDF 7897 kb)

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Supplementary Note

Preparation and characterization of synthetic standards 2 and 3 (PDF 1564 kb)

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Zha, L., Jiang, Y., Henke, M. et al. Colibactin assembly line enzymes use S-adenosylmethionine to build a cyclopropane ring. Nat Chem Biol 13, 1063–1065 (2017). https://doi.org/10.1038/nchembio.2448

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