G Protein-Coupled Receptor--Protein Interactions

This exciting volume elucidates how GPCRs organize signal transduction and control intracellular activities. Chapters written by the recognized expert in each respective topic offer state-of-the-art updates on the principles and technology, as well as stepwise protocols for methods currently applied to the analysis of protein-receptor interactions. Foreword (Robert J. Lefkowitz).

Series Preface (David R. Sibley).

Preface (Susan R. George & Brian F. O'Dowd).

Contributors.

1. Purification of the G Protein-Coupled Receptor Rhodopsin for Structural Studies (David Salom, Ning Li, Li Zhu, Izabela Sokal and Krzysztof Palczewski).

2. Cooperativity in Ligand Binding to Deduce Receptor/Receptor Interaction (Meritxell Vivo and Philip G. Strange).

3. Analysis of Receptor-Receptor Interaction Using Time-Resolved Fluorescence Resonance Energy Transfer (Douglas Ramsay and Graeme Milligan).

4. Real-Time Imaging of GPCR-Medicated Arresting Translocation as a Strategy to Evaluate Receptor-Protein Interactions (Robert H. Oakley, Larry S. Barak and Marc G. Caron).

5. Localization of G Protein Coupled Receptors and Interacting Proteins in Membrane Microdomains (K. Saidas Nair, Vijaya Narayanan and Vladlen Z. Slepak).

6. Identification of Peptide Ligand Binding Domains Within G Protein-Coupled Receptors Using Photoaffinity Labeling (Maoqing Dong and Laurence J. Miller).

7. Analysis in Real-Time of the Assembly and Disassembly of Soluble G Protein-Coupled Receptor-G Protein Complexes (Seam M. Biggs, Peter I. Simons, Anna Waller, Terry Founts, Tione Buranda, Eric R. Prossnitz and Larry A. Sklar).

8. Approaches to Studying Membrane Protein Oligomerization: Use of Cysteine Cross-Linking Strategies to Analyze Receptor-Receptor Interactions (Jonathan A. Javitch, Wen Guo and Marisa E. Ring).

9. Co-Immunoprecipitation as a Strategy to Evaluate Receptor-Receptor or Receptor-Protein Interactions (Randy A. Hall).

10. Use of MALDI-TOF and Mass Spectrometry in the Evaluation of Receptor-Protein Interactions (John R. Raymond, Henry Ayiku, Justin Turner, Andrew Gelasco and John M. Arthur).

11. Use of the Yeast Two-Hybrid System to Detect Receptor-Protein Interactions (Ridwan Lin, Nadine Kabbani, Alicia Binda, Victor Canfield and Robert Levenson).

12. GST Fusion Protein and Pull-down Strategies to Evaluate Receptor-Receptor or Receptor-Protein Interactions (Frank J.S. Lee and Fang Liu).

13. G-Protein-Coupled Fusion to Monitor Integral Membrane Protein-Protein Interactions (Henrik Dohlman and Kathleen Ehrhard).

14. Enzyme-Linked Immunosorbent Assays as Tools in Studying Interactions of G-Protein-Coupled Receptors (Torsten Schöneberg, Katrin Sangkuhl, Holger Römpler and Angela Schulz).

15. Electrophysiological Characterization of GPCR and Ligand-Gated Channel Crosstalk (Tak Pan Wong, Xian-Ming Yu and Yu Tian Wang).

16. BRET as a Strategy to Evaluate Receptor-Receptor or Receptor-Protein Interactions (Karen A. Eidne and Kevin D.G. Pfleger).

Index.

"…a good starting point for the investigation of complex GPCR-protein interactions, and covers a lot of ground by introducing many different methodologies." (E-STREAMS, September 2005)

" ... The Essentials of Trading will help those contemplating hands-on investing to determine whether they're cut out for the perilous and profitable world of global financial markets. If they proceed, Essentials will have armed them with the tricks for discipline and risk management." -- Rachel Koning Beals, SFO Magazine (July 2006)

SUSAN R. GEORGE, PhD, MD, FRCP, FACP, is Professor in the Departments of Medicine and Pharmacology at the University of Toronto and a Section Head of Molecular Pharmacology at the Centre for Addiction and Mental Health, Toronto.

BRIAN F. O'DOWD, PhD, is Professor in the Department of Pharmacology at the University of Toronto and a Section Head of Molecular Pharmacology at the Centre for Addiction and Mental Health, Toronto.

G PROTEIN-COUPLED RECEPTOR–PROTEIN INTERACTIONS

As the largest family of cell surface receptors, G protein-coupled receptors (GPCRs) mediate the signals from a diverse group of signaling molecules, including those responsible for the senses of smell, taste, and vision in mammals, as well as neurotransmission, hormone action, immune function, and numerous physiological functions in the brain and the periphery. The study of the GPCR signaling complex and signaling pathways has rapidly increased the number of proteins shown to interact with GPCRs. G Protein-Coupled Receptor–Protein Interactions offers a comprehensive guide to the variety of methodologies required to analyze receptor–receptor and receptor–protein interactions.

Featuring brief annotated bibliographies, clearly formatted protocols, and useful diagrams and illustrations, this highly accessible text covers a wide range of up-to-the-minute principles and technology, as well as protocols for methods currently applied to the analysis of receptor-protein interactions. Each chapter is written by an acknowledged expert, and each topic is clarified through a critical overview of key concepts.

Topics include:

• Bioluminescent resonance energy transfer
• Fluorescent resonance energy transfer
• Expression cloning
• Photoaffinity labeling
• Reconstitution
• Co-immunoprecipitation
• PDZ mutational analysis
• Confocal microscopy
• Dominant negative effect

Taken together, G Protein-Coupled Receptor–Protein Interactions will not only prove invaluable to scientists and academicians researching GPCR signal transduction systems, but will appeal to pharmaceutical industry professionals, since the GPCR family of proteins are targeted by over half of currently used therapeutic agents.