Receptor Signal Transduction Protocols

1. Front Matter

   Pages i-xiii

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2. Radioligand-Binding Methods for Membrane Preparations and Intact Cells

   • David B. Bylund, Jean D. Deupree, Myron L. Toews

   Pages 1-28

3. Determination of Allosteric Interactions Using Radioligand-Binding Techniques

   • Sebastian Lazareno

   Pages 29-46

4. Generation, Use, and Validation of Receptor-Selective Antibodies

   • John J. Mackrill

   Pages 47-65

5. Immunocytochemical Identification of G-Protein-Coupled Receptor Expression and Localization

   • Laura C. Mongan, Blair D. Grubb

   Pages 67-80

6. Generation and Use of Epitope-Tagged Receptors

   • R. A. Jeffrey McIlhinney

   Pages 81-98

7. Identification of G-Protein-Coupled Receptor mRNA Expression by Northern Blotting and In Situ Hybridization

   • Lucy F. Donaldson

   Pages 99-122

8. Electron Microscopic Studies of Receptor Localization

   • Rafael Luján

   Pages 123-136

9. Generation of Model Cell Lines Expressing Recombinant G-Protein-Coupled Receptors

   • Emmanuel Hermans

   Pages 137-153

10. Viral Infection Protocols

    • Antonio Porcellini, Antonio De Blasi

    Pages 155-166

11. Expression of G-Protein Signaling Components in Adult Mammalian Neurons by Microinjection

    • Stephen R. Ikeda

    Pages 167-181

12. Covalent Modification of G-Proteins by Affinity Labeling

    • Martin Hohenegger, Michael Freissmuth, Christian Nanoff

    Pages 183-195

13. Measurement of Agonist-Stimulated [35S]GTPγS Binding to Assess Total G-Protein and Gα-Subtype-Specific Activation by G-Protein-Coupled Receptors

    • Mark R. Dowling, Stefan R. Nahorski, R. A. John Challiss

    Pages 197-206

14. Identification and Quantitation of G-Protein α-Subunits

    • Ian Mullaney, Graeme Milligan

    Pages 207-224

15. Analysis of Function of Receptor-G-Protein and Receptor-RGS Fusion Proteins

    • Richard J. Ward, Graeme Milligan

    Pages 225-247

16. Assessment of Receptor Internalization and Recycling

    • Jennifer A. Koenig

    Pages 249-273

17. G-Protein-Coupled Receptor Phosphorylation and Palmitoylation

    • Andrew B. Tobin, Mark Wheatley

    Pages 275-281

18. Identification of G-Protein-Coupled Receptor Phosphorylation Sites by 2D Phosphopeptide Mapping

    • Andree Blaukat

    Pages 283-297

19. Ubiquitination of G-Protein-Coupled Receptors

    • Adriano Marchese, Jeffrey L. Benovic

    Pages 299-305

20. Receptor Mutagenesis Strategies for Examination of Structure-Function Relationships

    • Marion Blomenröhr, Henry F. Vischer, Jan Bogerd

    Pages 307-322

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.

From reviews of the first edition...
"...should be present in all cellular and molecular laboratories of the world."
-Cellular and Molecular Biology

"...enthusiastically recommend[ed]...should figure prominently in any laboratory involved in receptor biochemistry, molecular biology, and pharmacology."
-Trends in Pharmacological Sciences

• Department of Cell Physiology and Pharmacology, University of Leicester, Leicester, UK

  Gary B. Willars, R. A. John Challiss

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