G Protein-Coupled Receptor Screening Assays, Softcover reprint of the original 1st ed. 2015 Methods and Protocols Methods in Molecular Biology Series, Vol. 1272

 This volumeexplores the considerable efforts that have been directed towards the development of G Protein-Coupled Receptors (GPCR) screening assays in order to disclose GPCR acting compounds, elucidate signaling mechanisms or evaluate compound?s efficacy. New discoveries in the field, along with the widely recognized need for better and safer pharmaceutical drugs constitute the main motivation for this book. Readers, both beginners and experienced researchers, will receive an updated overview of not only the established, but also the innovative technologies that promise to advance GPCR drug research. This book is organized into two major parts: the introductory part discusses the necessary foundations for the understanding of GPCR action and the rationale behind the design of the available screening assays; and part two provides detailed protocols for different screening approaches. Written in the highly successful Methods in Molecular Biology series format, the chapters include the kind of detailed description and implementation advice that is crucial for getting optimal results in the laboratory.

Practical and innovative, G Protein-Coupled Receptor Screening Assays: Methods and Protocols reaches out to everyone involved in the discovery of GPCR-active drugs, and provides a transversal overview of the different levels of GPCR signaling addressable in the different screening strategies and presents practical examples of how current assay technologies are contributing to new paradigms in GPCR drug research.

1. G-Protein Coupled Receptors: An Overview of Signaling Mechanisms and Screening Assays

Duarte Miguel F. Prazeres and Sofia Aires M. Martins

Part II: GPCR Screening Assays

2. Time–Resolved FRET Strategy to Screen GPCR Ligands Library
Nadia Oueslati, Candide Hounsou, Abderazak Belhocine, Thieric Rodriguez, Elodie Dupuis, Jurriaan M. Zwier, Eric Trinquet, Jean-Philippe Pin, and Thierry Durroux

3. Homogeneous Fluorescence Anisotropy-Based Assay for Characterization of Ligand Binding Dynamics to GPCRs in Budded Baculoviruses: The Case of Cy3B-NDP-a-MSH Binding to MC4 Receptors
Santa Veiksina, Sergei Kopanchuk, Olga Mazina, Reet Link, Anne Lille, and Ago Rinken

4. Construction of Recombinant HEK293 Cell Lines for the Expression of the Neurotensin Receptor NTSR1
Su Xiao, Joseph Shiloach, and Reinhard Grisshammer

5. cAMP Assay for GPCR Ligand Characterization: Application of BacMam Expression System
Olga Mazina, Anni Allikalt, Annika Heinloo, Reet Reinart-Okugbeni, Sergei Kopanchuk, and Ago Rinken

6. Ca²⁺ Mobilization Assays in GPCR Drug Discovery
Grzegorz Woszczek and Elisabeth Fuerst

7. Using Constitutive Activity to Define Appropriate High Throughput Screening Assays for Orphan G Protein-Coupled Receptors
Tony Ngo, James L. Coleman, and Nicola J. Smith

8. Monitoring G Protein-Coupled Receptor Activation Using the Protein Fragment Complementation Technique Split TEV
Michael C. Wehr, Sabrina Galinski, and Moritz J. Rossner

9. Quantifying GPCR Internalization: A Focus on the Kisspeptin Receptor
Macarena Pampillo and Andy V. Babwah

10. GPCR Oligomerization Analysis by Means of BRET and dFRAP
Francisco Ciruela and Víctor Fernández-Dueñas

11. Use of ImageJ to Recover Information from Individual Cells in a G-Protein Coupled Receptor Assay
J. R. C. Trabuco, Sofia Aires M. Martins, and Duarte Miguel F. Prazeres

12. Methods to Immobilize GPCR on the Surface of SPR Sensors
Laura Martínez-Muñoz, Rubén Barroso, Anabel Guedán Paredes, Mario Mellado, and José Miguel Rodríguez-Frade

13. Olfactory Receptor Screening Assay Using Nanovesicle-Immobilized Carbon Nanotube Transistor
Jong Hyunn Lim, Juhum Park, Seunghun Hong, and Tai Hyun Park

14. Label-Free Biosensor Assays in GPCR Screening
Manuel Grundmann and Evi Kostenis

15. Multi-Dimensional GPCR Profiling and Screening Using Impedance-Based Label-Free and Real-Time Assay
Ning Ke, Khanh Nguyen, Jeffrey Irelan, and Yama A. Abassi

16. Label-Free Functional Selectivity Assays
Ann M. Ferrie, Vasiliy Goral, Chaoming Wang, and Ye Fang

17. Measurement of Surface-Mediated Ca++ Transients on the Single-Cell Level in a Microfluidic Lab-on-a-chip Environment
Michael Kirschbaum, Magnus S. Jaeger, and Claus Duschl

18. Cell-Based Assays and Animal Models for GPCR Drug Screening
Hideo Takakura, Mitsuru Hattori, Miho Tanaka, and Takeaki Ozawa

19. Computer-Aided Design of GPCR Ligands
Hugo Gutiérrez-de-Terán, Henrik Keränen, Jhonny Azuaje, David Rodríguez, Johan Åqvist, Eddy Sotelo 

Includes cutting-edge methods and protocols

Provides step-by-step detail essential for reproducible results

Contains key notes and implementation advice

Includes supplementary material: sn.pub/extras