Receptor Binding Techniques

ISBN-13: 9781493959365 / Angielski / Miękka / 2016 / 309 str.

Anthony P. Davenport

Opis

The 3rd edition of this Methods in Molecular Biology(TM) book expands on classic and new methods and techniques for studying receptors in silico, in vitro and in vivo. Includes reagent and materials lists, laboratory protocols, troubleshooting tips and pitfalls.

Spis treści

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 Schülein

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 β-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 Johnström, Joseph L. Bird and Anthony P. Davenport

11. Dynamic In Vivo Imaging of Receptors in Small Animals using Positron Emission Tomography

Peter Johnström, 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

Blurb

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 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, Third Edition, aids scientists in continuing to study receptor binding.