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Structure and biosynthesis of a macrocyclic peptide containing an unprecedented lysine-to-tryptophan crosslink

Nature Chemistry volume 7, pages 431–437 (2015)Cite this article

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Abstract

Streptococcal bacteria use peptide signals as a means of intraspecies communication. These peptides can contain unusual post-translational modifications, providing opportunities for expanding our understanding of nature's chemical and biosynthetic repertoires. Here, we have combined tools from natural products discovery and mechanistic enzymology to elucidate the structure and biosynthesis of streptide, a streptococcal macrocyclic peptide. We show that streptide bears an unprecedented post-translational modification involving a covalent linkage between two unactivated carbons within the side chains of lysine and tryptophan. The biosynthesis of streptide was addressed by genetic and biochemical studies. The former implicated a new SPASM-domain-containing radical SAM enzyme StrB, while the latter revealed that StrB contains two [4Fe–4S] clusters and installs the unusual lysine-to-tryptophan crosslink in a single step. By intramolecularly stitching together the side chains of lysine and tryptophan, StrB provides a new route for biosynthesizing macrocyclic peptides.

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Figure 1: Biosynthetic gene cluster and structure of streptide.
Figure 2: Proposed biosynthesis for streptide.
Figure 3: StrB contains two [4Fe–4S] clusters.
Figure 4: StrB catalyses Lys-to-Trp crosslink formation in StrA.
Figure 5: Mechanistic model for Lys-to-Trp crosslink formation catalysed by StrB.

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Acknowledgements

The authors thank P. Güntert and H. Arthanari for invaluable advice on CYANA, I. Pelczer and K. Connover at the Princeton Chemistry NMR facility for assistance with NMR data acquisition, S. Booker for the gift of pDB1282, K. Shatalin for the gift of pKS1, and Z. Brown for advice regarding peptide synthesis. This work was supported by the National Institutes of Health (grant no. GM098299, to M.R.S.) and by Princeton University start-up funds.

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  1. Kelsey R. Schramma and Leah B. Bushin: These authors contributed equally to this work

Authors and Affiliations

  1. Department of Chemistry, Princeton University, Princeton, New Jersey, USA

    Kelsey R. Schramma, Leah B. Bushin & Mohammad R. Seyedsayamdost

  2. Department of Molecular Biology, Princeton University, Princeton, New Jersey, USA

    Mohammad R. Seyedsayamdost

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  1. Kelsey R. Schramma
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Contributions

K.R.S., L.B.B. and M.R.S. designed and executed experiments, and analysed data. M.R.S. conceived of the project and wrote the manuscript.

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Correspondence to Mohammad R. Seyedsayamdost.

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Schramma, K., Bushin, L. & Seyedsayamdost, M. Structure and biosynthesis of a macrocyclic peptide containing an unprecedented lysine-to-tryptophan crosslink. Nature Chem 7, 431–437 (2015). https://doi.org/10.1038/nchem.2237

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