Receptor Signal Transduction Protocols

ISBN-13: 9780896034952 / Angielski / Twarda / 1997 / 275 str.

R. A. John Challis

Opis

Gaining an understanding of the mechanisms by which cells process and respond to extracellular cues has become a major goal in many areas of bi- ogy and has attracted the attentions of almost every traditional discipline within the biological sciences. At the heart of these divergent endeavors are common methods that can aid biochemists, physiologists, and pharmacologists in ta- ling the specific questions addressed by their research. In Receptor Signal Transduction Protocols, a diverse array of meth- ologies employed to interrogate ligand--receptor and receptor-effector int- actions are described by authors who have devised and successfully applied them. The authors blend excellent descriptions and applications of fairly well established methodologies with new technologies at the cutting-edge of signal transduction research and as such I hope the present volume will complement and extend a previous excellent volume in this series edited by David Kendall and Stephen Hill (Methods Molecular Biology, vol. 41, Signal Transd- tion Protocols).

Spis treści

Methods for Characterizing Receptors. Radioligand-Binding Methods for Membrane Preparations and Intact Cells Mary Keen Site-Directed Mutagenesis and Chimeric Receptors in the Study of Receptor-Ligand Binding Mark E. Olah and Gary L. Stiles Approaches to the Stable Transfection of G Protein-Coupled Receptors Andrea Townsend-Nicholson Methods for Transient Expression of Hetero-Oligomeric Ligand-Gated Ion Channels Hartmut Lüddens and Esa R. Korpi The Generation of Receptor-Selective Antibodies John J. Mackrill In Situ Hybridization Judith C.W. Mak and Peter J. Barnes Immunocytochemical Methods for Investigating Receptor Localization Maria I. Fonseca and R. Dale Brown Methods for Studying Receptor Coupling Mechanisms. Measurement of Agonist-Stimulated [35S]GTPgS Binding to Cell Membranes Sebastian Lazareno Autoradiographic Visualization in Brain of Receptor-G Protein Coupling Using [35S]GTPgS Binding Laura J. Sim, Dana E. Selley, and Steven R. Childers Agonist-Induced High-Affinity GTP Hydrolysis as an Index of Receptor-Mediated G Protein Activation in Mammalian Brain Membranes Yuji Odagaki and Kjell Fuxe Use of Random-Saturation Mutagenesis to Study Receptor-G Protein Coupling Ethan S. Burstein, Tracy A. Spalding, and Mark R. Brann Identification and Quantitation of G Protein a-Subunits Ian Mullaney and Graeme Milligan Covalent Modification of G Proteins by Affinity Labeling Martin Hohenegger, Michael Freissmuth, and Christian Nanoff Heterologous Expression of Receptors and Signaling Proteins in Adult Mammalian Sympathetic Neurons by Microinjection Stephen R. Ikeda Nuclear Application of Antisense Oligonucleotides by Microinjection and Ballistomagnetic Transfer to Identity G Protein Heterotrimers Activating PhospholipaseC Frank Kalkbrenner, Edgar Dippel, Matthias Schroff, Burghardt Wittig, and Günter Schultz Use of Antisense-Generating Plasmids to Probe the Function of Signal Transduction Proteins in Primary Neurons Fe C. Abogadie, Yvonne Vallis, Noel J. Buckley, and Malcolm P. Caulfield Methods for Investigating the Regulation of Receptor Function Protocols Employed in the Investigation of G Protein-Coupled Receptor Phosphorylation Andrew B. Tobin Assay of G Protein-Coupled Receptor Kinase Activity by Rhodopsin Phosphorylation Alison W. Gagnon and Eamonn Kelly Radioligand Binding Measurements of Receptor Sequestration in Intact and Permeabilized Cells Diana M. Slowiejko and Stephen K. Fisher Regulation of Receptor Expression: Analysis of Receptor mRNA and Gene Transcription Philip A. Iredale, Maura E. Charlton, John D. Alvaro, and Ronald S. Duman Index.

Blurb

R. A. J. Challiss assembles in Receptor Signal Transduction Protocols an outstanding collection of state-of-the-art methods from leading experts for interrogating ligand-receptor and receptor-effector interactions. These powerful methods emphasize the regulation of G-protein-coupled receptors and focus on receptor characterization, methods for investigating how receptors couple to G-proteins, and approaches to studying how receptor expression and function is regulated. Designed to be easily reproducible, the protocols offered include chimeric receptor construction, antibody generation, transient and stable transfection methods, in situ hybridization, as well as immunocytochemistry, functional assays of G-protein function, antisense methods for defining receptor-effector coupling pathways, and many more.

Receptor Signal Transduction Protocols blends authoritative descriptions and applications of well-established methodologies with many novel techniques at the cutting edge of signal transduction research. The coverage cuts across disciplines and allows those already well-versed in one area to expand their repertoire of potent methods and broaden their research horizons. Every researcher interested in signal transduction today will find this richly practical guide indispensable.