title: Studying binding specificities of peptide recognition modules by high-throughput phage display selections.
abstract: Peptide recognition modules (PRMs) play critical roles in cellular processes, including differentiation, proliferation and cytoskeleton organization. PRMs normally bind to short linear motifs in protein ligands, and by so doing recruit proteins into signaling complexes. Based on the binding specificity profile of a PRM, one can predict putative natural interaction partners by searching genome databases. Candidate interaction partners can in turn provide clues to assemble potential in vivo protein complexes that the PRM may be involved with. Combinatorial peptide libraries have proven to be effective tools for profiling the binding specificities of PRMs. Herein, we describe high-throughput methods for the expression and purification of PRM proteins and the use of peptide-phage libraries for PRM specificity profiling. These high-throughput methods greatly expedite the study of PRM families on a genome-wide scale.
authors.affiliation: Donnelly Centre for Cellular and Biomolecular Research, University of Toronto, Toronto, ON, Canada.
authors.forename: Haiming
authors.initials: H
authors.lastname: Huang
authors.forename: Sachdev S
authors.initials: SS
authors.lastname: Sidhu
mesh_terms: D057166:High-Throughput Screening Assays; D019151:Peptide Library; D011485:Protein Binding; D060066:Protein Interaction Maps; D011506:Proteins / Q000032:analysis* / Q000378:metabolism*; D012641:Selection, Genetic
chemical_list: D019151:Peptide Library; D011506:Proteins