title: Engineering sortase A by screening a second-generation library using phage display.
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.
authors.affiliation: Interfaculty Institute of Biochemistry, University of Tübingen, Hoppe-Seyler-Str. 4, D-72076, Tübingen, Germany.
authors.forename: Lena
authors.initials: L
authors.lastname: Schmohl
authors.affiliation: Interfaculty Institute of Biochemistry, University of Tübingen, Hoppe-Seyler-Str. 4, D-72076, Tübingen, Germany.
authors.forename: Jan
authors.initials: J
authors.lastname: Bierlmeier
authors.affiliation: Institute of Chemistry and Biochemistry, Freie Universität Berlin, Thielallee 63, D-14195, Berlin, Germany.
authors.forename: Fabian
authors.initials: F
authors.lastname: Gerth
authors.affiliation: Institute of Chemistry and Biochemistry, Freie Universität Berlin, Thielallee 63, D-14195, Berlin, Germany.
authors.forename: Christian
authors.initials: C
authors.lastname: Freund
authors.affiliation: Interfaculty Institute of Biochemistry, University of Tübingen, Hoppe-Seyler-Str. 4, D-72076, Tübingen, Germany.
authors.forename: Dirk
authors.initials: D
authors.lastname: Schwarzer
mesh_terms: D019881:Aminoacyltransferases / Q000235:genetics / Q000378:metabolism*; D001426:Bacterial Proteins / Q000235:genetics / Q000378:metabolism*; D003546:Cysteine Endopeptidases / Q000235:genetics / Q000378:metabolism*; D009154:Mutation / Q000235:genetics; D019151:Peptide Library; D010455:Peptides / Q000235:genetics / Q000378:metabolism; D015202:Protein Engineering / Q000379:methods*; D013379:Substrate Specificity
chemical_list: D001426:Bacterial Proteins; D019151:Peptide Library; D010455:Peptides; D019881:Aminoacyltransferases; C120542:sortase A; D003546:Cysteine Endopeptidases
keywords: directed evolution; phage display; protein engineering; sortase A; sortase-mediated ligation