G Protein-Coupled Receptors in Drug Discovery (2^nd Ed., Softcover reprint of the original 2nd ed. 2015) Methods and Protocols Methods in Molecular Biology Series, Vol. 1335

This detailed volume provides an overview of recent techniques employed in the field of G protein-coupled receptors (GPCRs) to screen for new drugs and to derive information about their receptor structure, dynamics, and function for the purpose of developing improved therapeutics. Owing to remarkable recent advances in the structural, biophysical and biochemical analyses of these receptors, as well as a growing body of evidence hinting at the possible relevance of allosteric modulators, biased agonists and oligomer-selective ligands as improved therapeutic agents, drug discovery for GPCRs has recently taken a completely new direction. For this book, expert contributors have shared their protocols and views on the impact of these methodologies on modern drug discovery. Written for the highly successful Methods in Molecular Biology series, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols and tips on troubleshooting and avoiding known pitfalls.

Practical and fully updated, G Protein-Coupled Receptors in Drug Discovery: Methods and Protocols, Second Edition serves as an ideal guide for a diverse audience from structural and molecular biologists to pharmacologists and drug designers who wish to explore this extensive class of key drug targets.

1. Purification of Stabilized GPCRs for Structural and Biophysical Analyses

            James C. Errey, Andrew S. Doré, Andrei Zhukov, Fiona H. Marshall, and Robert M. Cooke

2. Purification and Crystallization of a Thermostabilized Agonist-Bound Conformation of the Human Adenosine A_2A Receptor

            Christopher G. Tate and Guillaume Lebon

3. 2D Projection Analysis of GPCR Complexes by Negative Stain Electron Microscopy

            Alys Peisley and Georgios Skiniotis

4. Nuts and Bolts of CF₃ and CH₃ NMR Towards the Understanding of Conformational Exchange of GPCRs

            R. Scott Prosser and Tae Hun Kim

5. Single-Molecule Fluorescence Microscopy for the Analysis of Fast Receptor Dynamics

            Julia Wagner, Titiwat Sungkaworn, Katrin G. Heinze, Martin J. Lohse, and Davide Calebiro

6. Quantitative Multi-Color Detection Strategies for Bioorthogonally-Labeled GPCRs

            Minyoung Park, He Tian, Saranga Naganathan, Thomas P. Sakmar, and Thomas Huber

7. Approaches to Characterize and Quantify Oligomerization of GPCRs

            Sara Marsango, María José Varela Liste, and Graeme Milligan

8. Monitoring G Protein Activation in Cells with BRET

            Ikuo Masuho, Kirill A. Martemyanov, and Nevin A. Lambert

9. Use of Fluorescence Indicators in Receptor Ligands

            Kaleeckal G. Harikumar and Laurence J. Miller

10. Detection and Quantification of Intracellular Signalling Using FRET-Based Biosensors and High Content Imaging

            Michelle L. Halls, Daniel P. Poole, Andrew M. Ellisdon, Cameron J. Nowell, and Meritxell Canals

11. The Measurement of Receptor Signaling

            Terry Kenakin

12. Approaches to Assess Functional Selectivity in GPCRs: Evaluating G Protein Signaling in an Endogenous Environment

            Laura M. Bohn, Lei Zhou, and Jo-Hao Ho

13. Bioluminescence Resonance Energy Transfer Approaches to Discover Bias in GPCR Signaling

            Elizabeth K.M. Johnstone and Kevin D.G. Pfleger

14. Virus-Mediated Expression of DREADDs for In Vivo Metabolic Studies

            Mario Rossi, Zhenzhong Cui, Ken-ichiro Nakajima, Jianxin Hu, Lu Zhu, and Jürgen Wess

15. High-Throughput Screening for Allosteric Modulators of GPCRs

            Robert L. Bertekap, Jr., Neil T. Burford, Zhuyin Li, and Andrew Alt

16. Radioligand Binding Assay for an Exon 11-Associated Mu Opioid Receptor Target

            Gina F. Marrone, Susruta Majumdar, and Gavril W. Pasternak

17. Docking and Virtual Screening Strategies for GPCR Drug Discovery

            Thijs Beuming, Bart Lenselink, Daniele Pala, Fiona McRobb, Matt Repasky, and Woody Sherman

18. The Dynamic Process of Drug-GPCR Binding at either Orthosteric or Allosteric Sites Evaluated by Metadynamics

            Sebastian Schneider, Davide Provasi, and Marta Filizola

19. Experiment-Guided Molecular Modeling of Protein-Protein Complexes Involving GPCRs

            Irina Kufareva, Tracy M. Handel, and Ruben Abagyan

20. Interaction Fingerprints and Their Applications to Identify Hot Spots

            Andrés F. Marmolejo, José L. Medina-Franco, Marc Giulianotti, and Karina Martinez-Mayorga

Includes cutting-edge methods and protocols on GPCRs in drug discovery

Provides step-by-step detail essential for reproducible results

Contains key notes and implementation advice from the experts