G Protein-Coupled Receptors : Essential Methods

by Editor: David Poyner (University of Aston); Editor: Mark Wheatley (University of Birmingham )

• ISBN13: 9780470749227
• ISBN10: 0470749229
• Format: eBook
• Copyright: 2009-12-01
• Publisher: Wiley

Summary

G-Protein Coupled Receptors (GPCRs) are not only the largest protein family in the human genome but are also the single biggest target for therapeutic agents. Research into GPCRs is therefore growing at a fast pace and the range of techniques that can be applied to GPCRs is vast and continues to grow. This book provides an invaluable bench-side guide into the best and most up-to-date techniques for current and future research on GPCRs.With contributions from leading international authorities, this book equips readers with clear and detailed protocols for both well-known and up-and-coming techniques along with hints and tips for success. All the methods have been tried and tested by leading international research labs and are presented in easy-to-follow stages along with a useful overview of each technique.This book is an essential resource for all researchers in molecular biology, biochemistry, pharmacology and for graduate students.

Table of Contents

Preface

List of Contributors

Measurement of Ligand G-protein-coupled Receptor Interactions Using Radioligand Binding Techniques

Introduction

Methods and Approaches

References

Second Messenger Assays for G-protein-coupled Receptors: cAMP, Ca²⁺, Inositol Phosphates, ERK1/2

Introduction

Methods and Approaches

Troubleshooting

References

Use of the [³⁵S]GTPgS Binding Assay to Determine Ligand Efficacy at G-protein-coupled Receptors

Introduction

Methods and Approaches

Troubleshooting

Acknowledgements

References

Quantitative Imaging of Receptor Trafficking

Introduction

Methods and Approaches

Troubleshooting

References

Production of Recombinant G-protein-coupled Receptor in Yeast for Structural and Functional Analysis

Introduction

Methods and Approaches

Troubleshooting

References

Monitoring GPCR Protein Complexes Using Bioluminescence Resonance Energy Transfer

Introduction

Methods and Approaches

Troubleshooting

References

Using Intramolecular Fluorescence Resonance Energy Transfer to Study Receptor Conformation

Introduction

Methods and Approaches

Troubleshooting

References

A Disulfide Cross-linking Strategy Useful for Studying Ligand-induced Structural Changes in GPCRs

Introduction

Methods and Approaches

General Considerations, Caveats and Troubleshooting

Acknowledgements

References

Use of Fluorescence Correlation Spectroscopy to Study the Diffusion of G-protein-coupled Receptors

Introduction

Methods and Approaches

Measuring the Diffusion of a Ligand-occupied GPCR

Data Analysis

Troubleshooting

References

Identification and Proteomic Analysis of GPCR Phosphorylation

Introduction

Methods

Acknowledgements

References

Measurement and Visualization of G-protein-coupled Receptor Trafficking by Enzyme-linked Immunosorbent Assay and Immunofluorescence

Introduction

Methods and Approaches

Troubleshooting

References

Substituted Cysteine Accessibility Method (SCAM)

Introduction

Methods and Approaches

Troubleshooting

References

Homology Modelling of G-protein-coupled Receptors

Introduction

Methods and Approaches

Troubleshooting

Automated Methods for Generating Models of GPCRs

References

Appendix: Site-directed Mutagenesis and Chimeras

Introduction

Why Mutagenesis?

Troubleshooting

Conclusion

References

Table of Contents provided by Publisher. All Rights Reserved.