title: Probing the recognition of post-translational modifications by combining sortase-mediated ligation and phage-assisted selection.
abstract: Reversible post-translational modifications (PTMs) are key regulators of protein function and modulate a multitude of protein-protein interactions in signal transduction networks. Here, we describe a strategy for determining the modification preferences of PTM-binding proteins with only minimal protein amounts that can be obtained by immunoprecipitation from mammalian cell lysates. This method bases on the combination of sortase-mediated ligation and phage-assisted selection strategies. This method can be used to analyze the type of modification that mediates the interaction as well as the influence of the amino acids flanking the modification sites. We have demonstrated the applicability of this method by probing the interaction of phosphorylated tyrosine and serine residues with their respective binding domains.
authors.affiliation: Department of Protein Chemistry, Leibniz-Institut für Molekulare Pharmakologie, Robert-Rössle-Strasse 10, 13125 Berlin, Germany.
authors.forename: Till
authors.initials: T
authors.lastname: Teschke
authors.forename: Bernhard
authors.initials: B
authors.lastname: Geltinger
authors.forename: Alexander
authors.initials: A
authors.lastname: Dose
authors.forename: Christian
authors.initials: C
authors.lastname: Freund
authors.forename: Dirk
authors.initials: D
authors.lastname: Schwarzer
mesh_terms: D000595:Amino Acid Sequence; D019881:Aminoacyltransferases / Q000737:chemistry*; D001426:Bacterial Proteins / Q000737:chemistry*; D001435:Bacteriophages / Q000235:genetics; D003546:Cysteine Endopeptidases / Q000737:chemistry*; D015335:Molecular Probes / Q000737:chemistry / Q000235:genetics*; D019151:Peptide Library; D010455:Peptides / Q000737:chemistry / Q000235:genetics; D010766:Phosphorylation; D011499:Protein Processing, Post-Translational / Q000235:genetics; D011506:Proteins / Q000737:chemistry; D012694:Serine / Q000737:chemistry; D014443:Tyrosine / Q000737:chemistry
chemical_list: D001426:Bacterial Proteins; D015335:Molecular Probes; D019151:Peptide Library; D010455:Peptides; D011506:Proteins; D014443:Tyrosine; D012694:Serine; D019881:Aminoacyltransferases; C120542:sortase A; D003546:Cysteine Endopeptidases