書誌事項

Receptor signal transduction protocols

edited by Gary B. Willars and R.A. John Challiss

(Methods in molecular biology / John M. Walker, series editor, 746)(Springer protocols)

Humana Press, c2011

3rd ed

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注記

Includes bibliographical references and index

内容説明・目次

内容説明

As our understanding of G protein-coupled receptor (GPCR) signal transduction continues to grow, we cannot help but be struck by the emerging complexity and the ability of this receptor superfamily to continually surprise us as new facets are discovered. In this third edition of Receptor Signal Transduction Protocols, the expert contributors have taken into account the constant evolution of the GPCR field and dealt with methods that allow researchers to address many of the important rising issues. Serving as a companion to the previous editions, this volume contains thorough revisions of some core chapters, the complete rewriting of others to encompass new technological developments since the publication of the prior editions, and the commissioning of brand new, cutting-edge chapters to expand on previous coverage. Written in the highly successful Methods in Molecular Biology (TM) series format, chapters include introductions to the respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls. Authoritative and up-to-date, Receptor Signal Transduction Protocols, Third Edition serves the scientific community with a broad collection of the current, practical methodologies in this highly promising field.

目次

Part I: Developing Models for Studying GPCRs 1. Inducible Expression of G Protein-Coupled Receptors in Transfected Cells Beryl Koener and Emmanuel Hermans 2. Using the Flp-InTM T-RexTM System to Regulate GPCR Expression Richard J. Ward, Elisa Alvarez-Curto, and Graeme Milligan 3. Viral Infection for GPCR Expression in Eukaryotic Cells Antonio Porcellini, Luisa Iacovelli, and Antonio De Blasi 4. Generation of Epitope-Tagged GPCRs Yan Huang and Gary B. Willars 5. The Use of Site-Directed Mutagenesis to Study GPCRs Alex C. Conner, James Barwell, David R. Poyner, and Mark Wheatley 6. Approaches to Study GPCR Regulation in Native Systems Jonathon M. Willets 7. Heterologous Expression of GPCRs in Fission Yeast John Davey and Graham Ladds Part II: Examining GPCR Expression and Agonist-Induced Covalent Modifications 8. Radioligand Binding Methods for Membrane Preparations and Intact Cells David B. Bylund and Myron L. Toews 9. Quantification of GPCR mRNA Using Real-Time RT-PCR Trond Brattelid and Finn Olav Levy 10. Determining Allosteric Modulator Mechanism of Action: Integration of Radioligand Binding and Functional Assay Data Christopher J. Langmead 11. Design and Use of Fluorescent Ligands to Study Ligand-Receptor Interactions in Single Living Cells Stephen J. Briddon, Barrie Kellam, and Stephen J. Hill 12. Examining Site-Specific GPCR Phosphorylation Adrian J. Butcher, Andrew B. Tobin, and Kok Choi Kong 13. Ubiquitination of GPCRs Adriana Caballero and Adriano Marchese Part III: Examining Early Events in GPCR Signaling 14. [35S]GTP S Binding as an Index of Total G-Protein and G -Subtype-Specific Activation by GPCRs Rajendra Mistry, Mark R. Dowling, and R.A. John Challiss 15. Using Calcium Imaging as a Readout of GPCR Activation Martin D. Bootman and H. Llewelyn Roderick 16. Measuring Spatio-Temporal Dynamics of Cyclic AMP Signaling in Real-Time Using FRET-Based Biosensors Frank Gesellchen, Alessandra Stangherlin, Nicoletta Surdo, Anna Terrin, Anna Zoccarato, and Manuela Zaccolo 17. Determining the Activation of Rho as an Index of Receptor Coupling to G12/13 Proteins Michio Nakaya, Mina Ohba, Motohiro Nishida, and Hitoshi Kurose 18. The Use of Translocating Fluorescent Biosensors for Real-Time Monitoring of GPCR-Mediated Signaling Events Carl P. Nelson and R.A. John Challiss Part IV: Receptor-Receptor and Receptor-Protein Interactions 19. Study of GPCR-Protein Interactions Using Gel Overlay Assays and Glutathione-S-Transferase-Fusion Protein Pull-Downs Ashley E. Brady, Yunjia Chen, Lee E. Limbird, and Qin Wang 20. Study of GPCR-Protein Interactions by BRET Martina Kocan and Kevin D.G. Pfleger 21. Time Resolved FRET Strategy with Fluorescent Ligands to Analyze Receptor Interactions in Native Tissues: Application to GPCR Oligomerization Martin Cottet, Laura Albizu, Laetitia Comps-Agrar, Eric Trinquet, Jean-Philippe Pin, Bernard Mouillac, and Thierry Durroux 22. Peptide Affinity Purification for the Isolation and Identification of GPCR-Associated Protein Complexes Pascal Maurice, Avais M. Daulat, and Ralf Jockers 23. Tandem Affinity Purification and Identification of GPCR-Associated Protein Complexes Avais M. Daulat, Pascal Maurice, and Ralf Jockers 24. Identification of GPCR Localization in Detergent Resistant Membranes Ranju Kumari and Anna Francesconi 25. Analysis of GPCR Localization and Trafficking James N. Hislop and Mark von Zastrow Part V: Statistical Methods 26. Statistical Methods in Research Domenico Spina

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