E-Book Overview
This collection of practical, cutting-edge techniques for the study of cell signaling provides detailed, step-by-step instructions, helpful notes, and troubleshooting tips that make even the most powerful of the newest techniques readily reproducible. The protocols presented include the use of peptide libraries to study transmembrane signaling; the use of single-cell assays to analyze signal transduction pathways; the reconstitution of signaling complexes; methods for analyzing protein-protein interactions, and more. Introductory reviews explain the basic theory and enable researchers new to the area to rapidly gain understanding, as well as command of the practical knowledge and expertise afforded by the protocols. Transmembrane Signaling Protocols makes available to all researchers the many state-of-the-art techniques that have recently led to landmark discoveries in transmembrane signaling.
E-Book Content
Peptide Recognition Mechanisms of Eukaryotic Signaling Modules Chi-Hon Lee, David Cowburn,
and John Kuriyan
1. Introduction The formation of specific protem-protein interactions is one of the key mechanisms for signal transduction mediated by tyrosme phosphorylation. These intermolecular mteracttons target signaling proteins to particular cellular locations and modulate the enzymatic activities that further propagate the signal. A dtstmctive characteristic of the pathways that are mitiated by tyrosme phosphorylation is that target recognition and catalytic activity are usually functions of separate domains within the signaling molecules that participate m these pathways. Each of the signalmg molecules contains one or more of a set of modular peptide-bmdmg domains that are responsible for generating protein-protein interactions. Such peptide-recognition domains are modular in both structural and functtonal respects: They are capable of folding correctly