Cutting the Gordian knot: early and complete amino acid sequence confirmation of class II lasso peptides by HCD fragmentation

Scott A Jarmusch, Ingo Feldmann, Bernhard Blank-Landeshammer, Carlos Cortés-Albayay, Jean Franco Castro, Barbara Andrews, Juan A Asenjo, Albert Sickmann, Rainer Ebel, Marcel Jaspars

Research output: Contribution to journalArticle

Abstract

Lasso peptides are a diverse class of ribosomally synthesized and post-translationally modified peptides (RiPPs). Their proteolytic and thermal stability alongside their growing potential as therapeutics has increased attention to these antimicrobial peptides. With the advent of genome mining, the discovery of RiPPs allows for the accurate prediction of putatively encoded structures, however, MSn experiments only provide partial sequence confirmation, therefore 2D NMR experiments are necessary for characterisation. Multiple MS/MS techniques were applied to two structurally characterized lasso peptides, huascopeptin and leepeptin, and one uncharacterized lasso peptide, citrulassin C, which was not isolable in sufficient quantity for NMR analysis. We have shown that MS2 can be used to elucidate the full amino acid sequences previously predicted with genome mining for this compound class. HCD was able to open the macrocycles and fragment the newly opened linear peptides, confirming the complete amino acid sequences of the characterised lasso peptides. In addition, to determine if this technique could be applied at the earliest stages of the isolation process, we targeted a lasso peptide found by genome mining, citrulassin C, and were able to fully elucidate the amino acid sequence using only MS2 from a semi-crude extract of Streptomyces huasconensis HST28T.

Original languageEnglish
Pages (from-to)772-779
Number of pages8
JournalThe Journal of antibiotics
Volume73
Early online date9 Sep 2020
DOIs
Publication statusE-pub ahead of print - 9 Sep 2020

Keywords

  • CYCLIC-PEPTIDES
  • MICROCIN J25
  • ANTAGONIST
  • BI-32169
  • DESERT
  • TOOL

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    Jarmusch, S. A., Feldmann, I., Blank-Landeshammer, B., Cortés-Albayay, C., Castro, J. F., Andrews, B., Asenjo, J. A., Sickmann, A., Ebel, R., & Jaspars, M. (2020). Cutting the Gordian knot: early and complete amino acid sequence confirmation of class II lasso peptides by HCD fragmentation. The Journal of antibiotics, 73, 772-779. https://doi.org/10.1038/s41429-020-00369-z