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Studying Binding Specificities of Peptide Recognition Modules by High-Throughput Phage Display Selections

Protocol
First Online:
Part of the Methods in Molecular Biology book series (MIMB, volume 781)

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.

Key words

Phage display High-throughput selection Peptide recognition modules Protein expression and purification ELISA DNA sequencing 
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Notes

Acknowledgments

This work was supported by grant from the CIHR (MOP-93684) to S.S.S. We thank Andreas Ernst for figure preparation.

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© Springer Science+Business Media, LLC 2011

Authors and Affiliations

  1. 1.Donnelly Centre for Cellular and Biomolecular ResearchUniversity of TorontoTorontoCanada

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