Engineering sortase A by screening a second-generation library using phage display

J Pept Sci. 2017 Jul;23(7-8):631-635. doi: 10.1002/psc.2980. Epub 2017 Feb 10.

Abstract

Sortase-mediated ligation is one of the most commonly used chemo-enzymatic techniques for the site-specific modification of proteins. We have established a new library of sortase mutants for directed evolution of sortase substrate selectivity. Phage display screens of this second-generation library yielded sortase mutants that ligate substrate proteins containing an APxTG or FPxTG recognition sequence instead of the canonical LPxTG sorting motif. These findings indicate that the second-generation sortase library is well suited for sortase engineering in order to increase the versatility of sortase-mediated ligation. Copyright © 2017 European Peptide Society and John Wiley & Sons, Ltd.

Keywords: directed evolution; phage display; protein engineering; sortase A; sortase-mediated ligation.

MeSH terms

  • Aminoacyltransferases / genetics
  • Aminoacyltransferases / metabolism*
  • Bacterial Proteins / genetics
  • Bacterial Proteins / metabolism*
  • Cysteine Endopeptidases / genetics
  • Cysteine Endopeptidases / metabolism*
  • Mutation / genetics
  • Peptide Library
  • Peptides / genetics
  • Peptides / metabolism
  • Protein Engineering / methods*
  • Substrate Specificity

Substances

  • Bacterial Proteins
  • Peptide Library
  • Peptides
  • Aminoacyltransferases
  • sortase A
  • Cysteine Endopeptidases