Library Science

This new edition combines updates of key chapters from the first edition with a large number of new key methodologies that have emerged more recently for studying G protein-coupled receptors (GPCRs) and events immediately downstream of their activation. The methods are focused primarily on events at the receptor level, including ligand binding, 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.

Authors: Willars, Gary B.; Challiss, R. A. John

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Table of Contents

1. Radioligand-Binding Methods for Membrane Preparations and Intact Cells (pp. 001-028)
   Bylund, David B.; Deupree, Jean D.; Toews, Myron L.
2. Determination of Allosteric Interactions Using Radioligand-Binding Techniques (pp. 029-046)
   Lazareno, Sebastian
3. Generation, Use, and Validation of Receptor-Selective Antibodies (pp. 047-066)
   Mackrill, John J.
4. Immunocytochemical Identification of G-Protein-Coupled Receptor Expression and Localization (pp. 067-080)
   Mongan, Laura C.; Grubb, Blair D.
5. Generation and Use of Epitope-Tagged Receptors (pp. 081-098)
   McIlhinney, R. A. Jeffrey
6. Identification of G-Protein-Coupled Receptor mRNA Expression by Northern Blotting and In Situ Hybridization (pp. 099-122)
   Donaldson, Lucy F.
7. Electron Microscopic Studies of Receptor Localization (pp. 123-136)
   Lujan, Rafael
8. Generation of Model Cell Lines Expressing Recombinant G-Protein-Coupled Receptors (pp. 137-154)
   Hermans, Emmanuel
9. Viral Infection Protocols (pp. 155-166)
   Porcellini, Antonio; Blasi, Antonio De
10. Expression of G-Protein Signaling Components in Adult Mammalian Neurons by Microinjection (pp. 167-182)
    Ikeda, Stephen R.
11. Covalent Modification of G-Proteins by Affinity Labeling (pp. 183-196)
    Hohenegger, Martin; Freissmuth, Michael; Nanoff, Christian
12. Measurement of Agonist-Stimulated [35S]GTPyS Binding to Assess Total G-Protein and Ga-Subtype-Specific Activation by G-Protein-Coupled Receptors (pp. 197-206)
    Dowling, Mark R.; Nahorski, Stefan R.; Challiss, R. A. John
13. Identification and Quantitation of G-Protein a-Subunits (pp. 207-224)
    Mullaney, Ian; Milligan, Graeme
14. Analysis of Function of Receptor–G-Protein and Receptor–RGS Fusion Proteins (pp. 225-248)
    Ward, Richard J.; Milligan, Graeme
15. Assessment of Receptor Internalization and Recycling (pp. 249-274)
    Koenig, Jennifer A.
16. G-Protein-Coupled Receptor Phosphorylation and Palmitoylation (pp. 275-282)
    Tobin, Andrew B.; Wheatley, Mark
17. Identification of G-Protein-Coupled Receptor Phosphorylation Sites by 2D Phosphopeptide Mapping (pp. 283-298)
    Blaukat, Andree
18. Ubiquitination of G-Protein-Coupled Receptors (pp. 299-306)
    Marchese, Adriano; Benovic, Jeffrey L.
19. Receptor Mutagenesis Strategies for Examination of Structure–Function Relationships (pp. 307-322)
    Blomenrohr, Marion; Vischer, Henry F.; Bogerd, Jan
20. Study of G-Protein-Coupled Receptor–Protein Interactions by Bioluminescence Resonance Energy Transfer (pp. 323-334)
    Kroeger, Karen M.; Eidne, Karin A.
21. Fluorescence Resonance Energy Transfer to Study Receptor Dimerization in Living Cells (pp. 335-352)
    Bader, Jurgen E.; Beck-Sickinger, Annette G.
22. Identification of Protein Interactions by Yeast Two-Hybrid Screening and Coimmunoprecipitation (pp. 353-370)
    Tanowitz, Michael; Zastrow, Mark von
23. Study of G-Protein-Coupled Receptor–Protein Interactions Using Gel Overlay Assays and Glutathione-S-Transferase–Fusion Protein Pull-Downs (pp. 371-378)
    Brady, Ashley E.; Wang, Qin; Limbird, Lee E.
24. Receptor Knock-Out and Knock-In Strategies (pp. 379-390)
    Matsuda, Ikuo; Aiba, Atsu
25. Statistical Methods in G-Protein-Coupled Receptor Research (pp. 391-414)
    Freeman, Pat; Spina, Domenico

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