G Protein-Coupled Receptors (eBook, PDF)

Essential Methods
Redaktion: Poyner, David; Wheatley, Mark

• Format: PDF

Produktbeschreibung

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.

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Produktdetails

• Verlag: John Wiley & Sons
• Seitenzahl: 310
• Erscheinungstermin: 28. November 2009
• Englisch
• ISBN-13: 9780470749227
• Artikelnr.: 37299289

Autorenporträt

Dr David R. Poyner, University of Aston, Birmingham, UK; School of Life and Health Sciences, Aston University, Birmingham, UK. Professor?Mark Wheatley, University of Birmingham, UK.

Inhaltsangabe

Preface.
Contributors.
1 Measurement of Ligand-G-Protein-coupled Receptor Interactions (Katie
Leach, Celine Valant, Patrick M. Sexton and Arthur Christopoulos).
1.1 Introduction.
1.2 Methods and Approaches.
References.
2 Second Messenger Assays for G-Protein-coupled Receptors: cAMP, Ca2+,
Inositol Phosphates, ERK1/2 (Karen J. Gregory, Patrick M. Sexton, Arthur
Christopoulos and Caroline A. Hick).
2.1 Introduction.
2.2 Methods and Approaches.
2.3 Troubleshooting.
References.
3 Use of the [35S]GTP¿S Binding Assay to Determine Ligand Efficacy at
G-protein-coupled Receptors (Elodie Kara and Philip G. Strange).
3.1 Introduction.
3.2 Methods and Approaches.
3.3 Troubleshooting.
Acknowledgements.
References.
4 Quantitative Imaging of Receptor Trafficking (Andy R. James, Takeo Awaji,
F. Anne Stephenson and Nicholas A. Hartell).
4.1 Introduction.
4.2 Methods and Approaches.
4.3 Troubleshooting.
References.
5 Production of Recombinant G-Protein-coupled Receptor in Yeast for
Structural and Functional Analysis (Richard A.J. Darby, Mohammed Jamshad,
Ljuban Grgic, William J. Holmes and Roslyn M. Bill).
5.1 Introduction.
5.2 Methods and Approaches.
5.3 Troubleshooting.
References.
6 Monitoring GPCR-Protein Complexes Using Bioluminescence Resonance Energy
Transfer (Werner C. Jaeger, Kevin D.G. Pfleger and Karin A. Eidne).
6.1 Introduction.
6.2 Methods and Approaches.
6.3 Troubleshooting.
References.
7 Using Intramolecular Fluorescence Resonance Energy Transfer to Study
Receptor Conformation (Cornelius Krasel and Carsten Hoffmann).
7.1 Introduction.
7.2 Methods and Approaches.
7.3 Troubleshooting.
References.
8 A Disulfide Cross-linking Strategy Useful for Studying Ligand-induced
Structural Changes in GPCRs (Jian Hua Li, Stuart D.C. Ward, Sung-Jun Han,
Fadi F. Hamdan and Jürgen Wess).
8.1 Introduction.
8.2 Methods and Approaches.
8.3 General Considerations, Caveats and Troubleshooting.
Acknowledgements.
References.
9 Use of Fluorescence Correlation Spectroscopy to Study the Diffusion of
G-protein-coupled Receptors (Stephen J. Briddon, Jonathan A. Hern and
Stephen J. Hill).
9.1 Introduction.
9.2 Methods and Approaches.
9.3 Troubleshooting.
References.
10 Identification and Proteomic Analysis of GPCR Phosphorylation (Kok Choi
Kong, Sharad C. Mistry and Andrew B. Tobin).
10.1 Introduction.
10.2 Methods.
Acknowledgements.
References.
11 Measurement and Visualization of G-Protein-coupled Receptor Trafficking
by Enzyme-linked Immunosorbent Assay and Immunofluorescence (Stuart J.
Mundell, Shaista P. Nisar and Eamonn Kelly).
11.1 Introduction.
11.2 Methods and Approaches.
11.3 Troubleshooting.
References.
12 Substituted Cysteine Accessibility Method (SCAM) (George Liapakis and
Jonathan A. Javitch).
12.1 Introduction.
12.2 Methods and Approaches.
12.3 Troubleshooting.
References.
13 Homology Modelling of G-Protein-coupled Receptors (John Simms).
13.1 Introduction.
13.2 Methods and Approaches.
13.3 Troubleshooting.
13.4 Automated Methods for Generating Models of GPCRs.
References.
Appendix: Site-directed Mutagenesis and Chimeras (Alex Conner, Mark
Wheatley and David Poyner).
A.1 Introduction.
A.2 Why Mutagenesis?
A.3 Troubleshooting.
A.4 Conclusion.
References.
Index.