An Orthogonal D2O-Based Induction System that Provides Insights into d-Amino Acid Pattern Formation by Radical S-Adenosylmethionine Peptide Epimerases

Brandon I. Morinaka, Marjan Verest, Michael F. Freeman, Muriel Gugger, Jörn Piel

Research output: Contribution to journalArticlepeer-review

29 Scopus citations

Abstract

Radical S-adenosyl methionine peptide epimerases (RSPEs) are an enzyme family that accomplishes regiospecific and irreversible introduction of multiple d-configured residues into ribosomally encoded peptides. Collectively, RSPEs can generate diverse epimerization patterns in a wide range of substrates. Previously, the lack of rapid methods to localize epimerized residues has impeded efforts to investigate the function and applicative potential of RSPEs. An efficient mass spectrometry-based assay is introduced that permits characterization of products generated in E. coli. Applying this to a range of non-natural peptide-epimerase combinations, it is shown that the d-amino acid pattern is largely but not exclusively dictated by the core peptide sequence, while the epimerization order is dependent on the enzyme-leader pair. RSPEs were found to be highly promiscuous, which allowed for modular introduction of peptide segments with defined patterns.

Original languageEnglish (US)
Pages (from-to)762-766
Number of pages5
JournalAngewandte Chemie - International Edition
Volume56
Issue number3
DOIs
StatePublished - Jan 16 2017

Bibliographical note

Funding Information:
We thank M. Helf for a sample of PoyH and MS annotation software, and S. Fuchs for insightful discussion. J.P. acknowledges funding from the SNF (31003A_146992/1) and EU (SYNPEPTIDE), B.I.M. is a fellowship recipient of the Alexander von Humboldt foundation. M.F.F. is recipient of a Human Frontier Science Program Long-term Fellowship.

Publisher Copyright:
© 2017 Wiley-VCH Verlag GmbH & Co. KGaA, Weinheim

Keywords

  • epimerases
  • labeling
  • peptide biosynthesis
  • post-translation
  • radical-SAM

Fingerprint

Dive into the research topics of 'An Orthogonal D2O-Based Induction System that Provides Insights into d-Amino Acid Pattern Formation by Radical S-Adenosylmethionine Peptide Epimerases'. Together they form a unique fingerprint.

Cite this