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

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• 作者：Dr. Brandon I. Morinaka ; Marjan Verest ; Dr. Michael F. Freeman ; Dr. Muriel Gugger and Prof. Dr. Jö ; rn Piel
• 刊名：Angewandte Chemie
• 出版年：2017
• 出版时间：January 16, 2017
• 年：2017
• 卷：129
• 期：3
• 页码：780-784
• 全文大小：3115K
• ISSN：1521-3757

文摘

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.