This second edition of Receptor Signal Transduction Protocols not only has a new editor, but also a greater focus on G-protein-coupled receptors, their properties per se, and their coupling to immediate downstream binding partners―principally, although not exclusively, the heterotrimeric G-proteins. The new edition combines updates of key chapters from the first edition, as well as a large number of new contributions covering key methodologies that have emerged, or been extended to receptor/G-protein research, in the past 5–6 years. In common with many fields, the range of methods used to assess the first steps in signal transduction are continually expanding and methods that might have been considered too specialized five years ago are now sufficiently routine to be included here. Unlike many research areas, where off-the-shelf kits have made research basically foolproof, signal transduction research still requires considerable expertise, and the methods included here are provided by internationally recognized experts in their fields who have many years of experience using the methods they describe. This not only allows each chapter to impart a clear description of the method, but also to furnish invaluable troubleshooting advice for when things do not go entirely according to plan. Once again we would like to thank the Series Editor, John Walker, for the invitation to compile this second edition, and to express our gratitude to all of the authors who have enthusiastically agreed to provide the uniformly excellent contributions.
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In this second edition of a widely appreciated work, Receptor Signal Transduction Protocols, a panel of internationally recognized investigators presents their best methods for studying G-protein-coupled receptors (GPCRs) and events immediately downstream of their activation. This new edition combines updates of key chapters from the first edition with a large number of new contributions on the many successful methodologies that have emerged more recently. The methods are focused primarily on events at the receptor level, including ligand binding, on the genetic manipulation of receptors, the generation of model cell lines in which to study them, and the interaction and activation of G-proteins. Additional methods concentrate on receptor expression and localization, receptor internalization and post-translational modification, GPCR-protein interactions, and the use of knock-out and knock-in strategies for determining the physiological roles of receptors. The laboratory protocols follow the successful Methods in Molecular Biology™ series format, each one offering step-by-step laboratory instructions, an introduction outlining the principle behind the technique, lists of equipment and reagents, and tips on troubleshooting and avoiding known pitfalls.
Comprehensive and highly practical, Receptor Signal Transduction Protocols, Second Edition offers both novice and experienced investigators powerful cutting-edge techniques that provide an array of approaches and specific methods that will aid in the understanding of GPCR structure and function.
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