Mechanistic Investigations of PoyD, a Radical S-Adenosyl-l-methionine Enzyme Catalyzing Iterative and Directional Epimerizations in Polytheonamide A Biosynthesis.
(2018) Journal of the American chemical society — Vol. 140, n° 7, p. 2469-2477 (2018)
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- Authors
Berteau, Olivier 
- Abstract
- Ribosomally synthesized and post-translationally modified peptides (RiPPs) are a growing family of bioactive peptides. Among RiPPs, the bacterial toxin polytheonamide A is characterized by a unique set of post-translational modifications catalyzed by novel radical S-adenosyl-l-methionine (SAM) enzymes. Here we show that the radical SAM enzyme PoyD catalyzes in vitro polytheonamide epimerization in a C-to-N directional manner. By combining mutagenesis experiments with labeling studies and investigating the enzyme substrate promiscuity, we deciphered in detail the mechanism of PoyD. We notably identified a critical cysteine residue as a likely key H atom donor and demonstrated that PoyD belongs to a distinct family of radical SAM peptidyl epimerases. In addition, our study shows that the core peptide directly influences the epimerization pattern allowing for production of peptides with unnatural epimerization patterns.
- Affiliations
INRAe ChemSyBio
Citations
Parent, A., Benjdia, A., Guillot, A., Kubiak, X., Balty, C., Lefranc, B., Leprince, J., & Berteau, O. (2018). Mechanistic Investigations of PoyD, a Radical S-Adenosyl-l-methionine Enzyme Catalyzing Iterative and Directional Epimerizations in Polytheonamide A Biosynthesis. Journal of the American chemical society, 140(7), 2469-2477. https://doi.org/10.1021/jacs.7b08402 (Original work published 2018)