Stuffed Methyltransferase Catalyzes the Penultimate Step of Pyochelin Biosynthesis

Trey A. Ronnebaum, Jeffrey S. McFarlane, Thomas E. Prisinzano, Squire J. Booker, Audrey L. Lamb

Research output: Contribution to journalArticlepeer-review

9 Scopus citations

Abstract

Nonribosomal peptide synthetases use tailoring domains to incorporate chemical diversity into the final natural product. A structurally unique set of tailoring domains are found to be stuffed within adenylation domains and have only recently begun to be characterized. PchF is the NRPS termination module in pyochelin biosynthesis and includes a stuffed methyltransferase domain responsible for S-adenosylmethionine (AdoMet)-dependent N-methylation. Recent studies of stuffed methyltransferase domains propose a model in which methylation occurs on amino acids after adenylation and thiolation rather than after condensation to the nascent peptide chain. Herein, we characterize the adenylation and stuffed methyltransferase didomain of PchF through the synthesis and use of substrate analogues, steady-state kinetics, and onium chalcogen effects. We provide evidence that methylation occurs through an SN2 reaction after thiolation, condensation, cyclization, and reduction of the module substrate cysteine and is the penultimate step in pyochelin biosynthesis.

Original languageEnglish
Pages (from-to)665-678
Number of pages14
JournalBiochemistry
Volume58
Issue number6
DOIs
StatePublished - Feb 12 2019

Bibliographical note

Funding Information:
Research reported in this publication was made possible by the funds from The National Science Foundation (CHE-1403293 to A.L.L.) and The National Institute of General Medical Sciences of The National Institutes of Health (P20 GM103418 and R01 GM127655 to A.L.L., P20 GM113117 to T.E.P., and GM122595 to S.J.B.). T.A.R. and J.S.M. were supported by the National Institutes of Health Graduate Training Program in the Dynamic Aspects of Chemical Biology (T32 GM008545). J.S.M. was supported by an American Heart Association Predoctoral Fellowship (PRE33960374).

Funding Information:
*E-mail: lamb@ku.edu. Phone: (785) 864-5075. ORCID Thomas E. Prisinzano: 0000-0002-0649-8052 Squire J. Booker: 0000-0002-7211-5937 Audrey L. Lamb: 0000-0002-2352-2130 Funding Research reported in this publication was made possible by the funds from The National Science Foundation (CHE-1403293 to A.L.L.) and The National Institute of General Medical Sciences of The National Institutes of Health (P20 GM103418 and R01 GM127655 to A.L.L., P20 GM113117 to T.E.P., and GM122595 to S.J.B.). T.A.R. and J.S.M. were supported by the National Institutes of Health Graduate Training Program in the Dynamic Aspects of Chemical Biology (T32 GM008545). J.S.M. was supported by an American Heart Association Predoctoral Fellowship (PRE33960374). Notes The authors declare no competing financial interest.

Publisher Copyright:
© 2018 American Chemical Society.

ASJC Scopus subject areas

  • Biochemistry

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