Title Receptor signal transduction protocols / edited by Gary B. Willars, R.A. John Challiss

Edition 2nd ed Published Totowa, N.J. : Humana Press, ©2004

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Description 1 online resource (xiii, 418 pages) : illustrations Series Methods in molecular biology, 1064-3745 ; 259 Methods in molecular biology (Clifton, N.J.) ; v. 259 Contents Radioligand-binding methods for membrane preparations and intact cells / D.B. Bylund, J.D. Deupree and M.L. Toews -- Determination of allosteric interactions using radioligand-binding techniques / S. Lazareno -- Generation, use, and validation of receptor-selective antibodies / J.J. Mackrill -- Immunocytochemical identification of G-protein-coupled receptor expression and localization / L.C. Mongan and B.D. Grubb -- Generation and use of epitope-tagged receptors / R.A. McIlhinney -- Identification of G-protein-coupled receptor mRNA expression by northern blotting and in situ hybridization / L.F. Donaldson -- Electron microscopic studies of receptor localization / R. Lujan -- Generation of model cell lines expressing recombinant G-protein-coupled receptors / E. Hermans -- Viral infection protocols / A. Porcellini and A. De Blasi -- Expression of G-protein signaling components in adult mammalian neurons by microinjection / S.R. Ikeda -- Covalent modification of G-proteins by affinity labeling / M. Hohenegger, M. Freissmuth and C. Nanoff -- Measurement of agonist-stimulated [35S]GTP[gamma]S binding to assess total G-protein and G[alpha]-subtype-specific activation by G-protein-coupled receptors / M.R. Dowling, S.R. Nahorski and R.A. Challiss -- Identification and quantitation of G-protein [alpha]-subunits / I. Mullaney and G. Milligan -- Analysis of function of receptor-G-protein and receptor-RGS fusion proteins / R.J. Ward and G. Milligan -- Assessment of receptor internalization and recycling / J.A. Koenig -- G-protein-coupled receptor phosphorylation and palmitoylation / A.B. Tobin and M. Wheatley -- Identification of G-protein-coupled receptor phosphorylation sites by 2d phosphopeptide mapping / A. Blaukat -- Ubiquitination of G-protein-coupled receptors / A. Marchese and J.L. Benovic -- Receptor mutagenesis strategies for examination of structure-function relationships / M. Blomenrohr, H.F. Vischer and J. Bogerd -- Study of G-protein-coupled receptor-protein interactions by bioluminescence resonance energy transfer / K.M. Kroeger and K.A. Eidne -- Fluorescence resonance energy transfer to study receptor dimerization in living cells / J.E. Bader and A.G. Beck-Sickinger -- Identification of protein interactions by yeast two-hybrid screening and coimmunoprecipitation / M. Tanowitz and M. von Zastrow -- Study of G-protein-coupled receptor-protein interactions using gel overlay assays and glutathione-s-transferase-fusion protein pull-downs / A.E. Brady, Q. Wang and L.E. Limbird -- Receptor knock-out and knock-in strategies / I. Matsuda and A. Aiba -- Statistical methods in G-protein-coupled receptor research / P. Freeman and D. Spina Summary 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 Bibliography Includes bibliographical references and index Notes Print version record Subject Cell receptors -- Research -- Methodology Cellular signal transduction -- Research -- Methodology Signal Transduction -- physiology SCIENCE -- Life Sciences -- Biochemistry. Cell receptors -- Research -- Methodology Cellular signal transduction -- Research -- Methodology Genre/Form Laboratory manuals Laboratory manuals. Manuels de laboratoire. Form Electronic book Author Willars, Gary B Challiss, R. A. J. (R. A. John) LC no. 2003027534 ISBN 9781592597543 1588293297 9781588293299 1592597548

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