Receptor Binding Techniques Methods and Protocols

About this book

A broad definition of a receptor is a specialized protein on or in a cell that recognizes and binds a specific ligand to undergo a conformational change, leading to a physiological response or change in cell function. A ligand can be an endogenous neurotransmitter, hormone, paracrine/autocrine factor, or a synthetic drug that may function as an agonist or antagonist. The third edition of Receptor Binding Techniques: Methods and Protocols expands upon the methods and techniques used for studying receptors in silico, in vitro and in vivo. Comprehensive chapters describe how to use online resources for experimental research such as prediction of receptor-ligand interactions and mine the IUPHAR receptor database.

Classical techniques of radioligand binding, quantitative autoradiography and their analyses are complemented by the use of immunocytochemistry for the cellular localization of receptor protein and hybridization to detect receptor mRNA. Protocols using fluorescent labeled ligands are described to visualise receptors in living cells, their interaction with beta-arrestin to measure ligand-induced internalisation and green fluorescent protein to study trafficking. Non-radioactive, chemiluminescent cAMP and arrestin assays facilitate the identification of novel 'biased agonists'.

Detailed methods are provided for in vivo imaging of receptors using positron emission tomography (PET). Written in the highly successful Methods in Molecular Biology series format, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and key tips on troubleshooting and avoiding known pitfalls. Authoritative and practical, Receptor Binding Techniques: Methods and Protocols, Third Edition, aids scientists in continuing to study receptor binding.

Contents

1. Receptor Databases and Computational Websites for Ligand Binding Brinda K. Rana, Philip E. Bourne and Paul A. Insel
2. How to use the IUPHAR Receptor Database to Navigate Pharmacological Data Chidochangu P. Mpamhanga, Joanna L. Sharman, Anthony J. Harmar, and NC-IUPHAR
3. Radioligand Binding Assays and their Analysis Janet J. Maguire, Rhoda E. Kuc and Anthony P. Davenport
4. Use of Scintillation Proximity Assay (SPA) to Measure Radioligand Binding to Immobilised Receptors without Separation of Bound from Free Ligand Jenny Berry, Molly Price-Jones, and Barbara Killian
5. Visualisation and Analysis of Vascular Receptors using Confocal Laser Scanning Microscopy and Fluorescent Ligands Craig J Daly, Ingela Parmryd and John C. McGrath
6. Dissecting the Pharmacology of G Protein Coupled Receptor Signalling Complexes using Bimolecular Fluorescence Complementation Laura E Kilpatrick and Nicholas D Holliday
7. Live Cell Imaging of G Protein-Coupled Receptors Anke Teichmann, Antje Schmidt , Burkhard Wiesner , Alexander Oksche , and Ralf Schulein
8. Characterization of G-protein Coupled Receptor (GPCR) Modulators using Homogeneous cAMP Assays Daniel L. Bassoni, Qumber Jafri, Sunitha Sastry, Mahesh Mathrubutham and Tom Wehrman
9. Measurements of beta-arrestin Recruitment to Activated Seven Transmembrane Receptors using Enzyme Complementation Daniel L. Bassoni, William J. Raab, Philip L. Achacoso, Chin Yee Loh and Tom S. Wehrman
10. Quantitative Phosphor Imaging Autoradiography of Radioligands for Positron Emission Tomography Peter Johnstrom, Joseph L. Bird and Anthony P. Davenport
11. Dynamic In Vivo Imaging of Receptors in Small Animals using Positron Emission Tomography Peter Johnstrom, Tim D. Fryer, Joseph L. E. Bird, Hugh K. Richards, and Anthony P. Davenport
12. Cellular Localization of Receptors using Antibodies Visualised by Light and Dual Labelling Confocal Microscopy Anthony P. Davenport and Rhoda E. Kuc
13. Detection of mRNA Encoding Receptors by in Situ And Northern Hybridisation Alessandra P. Princivalle, Rachel M. C Parker, Terri J. Dover, and Nicholas M. Barnes

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