Willars / Challiss Receptor Signal Transduction Protocols

2. Auflage 2004
ISBN: 978-1-59259-754-3
Verlag: Humana Press
Format: PDF
Kopierschutz: 1 - PDF Watermark

E-Book, Englisch, Band 259, 418 Seiten, eBook

Reihe: Methods in Molecular Biology

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.

Zielgruppe

Research

Autoren/Hrsg.

Willars, Gary B.

Challiss, R. A. John

Inhaltsverzeichnis

Radioligand-Binding Methods for Membrane Preparations and Intact Cells.- Determination of Allosteric Interactions Using Radioligand-Binding Techniques.- Generation, Use, and Validation of Receptor-Selective Antibodies.- Immunocytochemical Identification of G-Protein-Coupled Receptor Expression and Localization.- Generation and Use of Epitope-Tagged Receptors.- Identification of G-Protein-Coupled Receptor mRNA Expression by Northern Blotting and In Situ Hybridization.- Electron Microscopic Studies of Receptor Localization.- Generation of Model Cell Lines Expressing Recombinant G-Protein-Coupled Receptors.- Viral Infection Protocols.- Expression of G-Protein Signaling Components in Adult Mammalian Neurons by Microinjection.- Covalent Modification of G-Proteins by Affinity Labeling.- Measurement of Agonist-Stimulated [35S]GTP?S Binding to Assess Total G-Protein and G?-Subtype-Specific Activation by G-Protein-Coupled Receptors.- Identification and Quantitation of G-Protein ?-Subunits.- Analysis of Function of Receptor-G-Protein and Receptor-RGS Fusion Proteins.- Assessment of Receptor Internalization and Recycling.- G-Protein-Coupled Receptor Phosphorylation and Palmitoylation.- Identification of G-Protein-Coupled Receptor Phosphorylation Sites by 2D Phosphopeptide Mapping.- Ubiquitination of G-Protein-Coupled Receptors.- Receptor Mutagenesis Strategies for Examination of Structure-Function Relationships.- Study of G-Protein-Coupled Receptor-Protein Interactions by Bioluminescence Resonance Energy Transfer.- Fluorescence Resonance Energy Transfer to Study Receptor Dimerization in Living Cells.- Identification of Protein Interactions by Yeast Two-Hybrid Screening and Coimmunoprecipitation.- Study of G-Protein-Coupled Receptor-Protein Interactions Using Gel Overlay Assaysand Glutathione-S-Transferase-Fusion Protein Pull-Downs.- Receptor Knock-Out and Knock-In Strategies.- Statistical Methods in G-Protein-Coupled Receptor Research.