G Protein-Coupled Receptor Signaling, 1st ed. 2019 Methods and Protocols Methods in Molecular Biology Series, Vol. 1947

Part I: Probing Structural Properties of GPCRs

1. Assessment of Conformational State Transitions of Class B GPCRs Using Molecular Dynamics

            Chenyi Liao, Victor May, and Jianing Li

2. Molecular Dynamic Simulations to Probe Water Permeation Pathways of GPCRs

            Katsufumi Tomobe, Eiji Yamamoto, and Kenji Yasuoka

3. Expression and Purification of a Functional E. coli ¹³CH₃-Methionine-Labeled Thermostable Neurotensin Receptor 1 Variant for Solution NMR Studies

4. A Combined Cell-Free Protein Synthesis and Fluorescence-Based Approach to Investigate GPCR Binding Properties

            Anne Zemella, Theresa Richter, Lena Thoring, and Stefan Kubick

Part II: Tweaking Ligands for GPCR Studies

5. Furan Cross-Linking Technology for Investigating GPCR-Ligand Interactions

            Marleen Van Troys, Willem Vannecke, Christophe Ampe, and Annemieke Madder

6. Optical Regulation of Class C GPCRs by Photoswitchable Orthogonal Remotely-Tethered Ligands

            Amanda Acosta-Ruiz, Johannes Broichhagen, and Joshua Levitz

7. Chemoselective Acylation of Hydrazinopeptides to Access Fluorescent Probes for Time-Resolved FRET Assay on GPCRs

            Sridévi M. Ramanoudjame, Lucie Esteoulle, Stéphanie Riché, Jean-François Margathe, Thierry Durroux, Iuliia A. Karpenko, and Dominique Bonnet

Part III: Bioluminescence and Fluorescence Resonance Transfer Energy Approaches

8. Time-Resolved FRET-Based Assays to Characterize G Protein-Coupled Receptor Hetero-Oligomer Pharmacology

            Joyce Heuninck, Candide Hounsou, Elodie Dupuis, Eric Trinquet, Bernard Mouillac, Jean-Philippe Pin, Dominique Bonnet, and Thierry Durroux

9. Combining Conformational Profiling of GPCRs with CRISPR/Cas9 Gene Editing Approaches

            Kyla Bourque, Dominic Devost, Asuka Inoue, and Terence E. Hébert

10. Measuring GPCR Stoichiometry Using Types -1, -2 and -3 Bioluminescence Resonance Energy Transfer-Based Assays

            James H. Felce, John R. James, and Simon J. Davis

11. Combining SRET² and BiFC to Study GPCR Heteromerization and Protein-Protein Interactions

            Amina M. Bagher, Melanie E.M. Kelly, and Eileen M. Denovan-Wright

12. Quantification and Comparison of Signals Generated by Different FRET-Based cAMP Reporters

            Andreas Koschinski and Manuela Zaccolo

Part IV: Exploring GPCR Signaling Properties

13. Measuring GPCR-Induced Activation of Protein Tyrosine Phosphatases (PTP) Using In-Gel and Colorimetric PTP Assays

            Geneviève Hamel-Côté, Fanny Lapointe, and Jana Stankova

14. Measurement of β-Arrestin Recruitment at GPCRs Using the Tango Assay

            Geneviève Laroche and Patrick M. Giguère

15. Probing the Interactome of Corticotropin-Releasing Factor Receptor Heteromers Using Mass Spectrometry

Burcu Hasdemir, Juan A. Oses-Prieto, Alma Burlingame, and Aditi Bhargava

Part V: Visualizing Subcellular Compartmentalization of GPCRs

16. Monitoring the Aggregation of GPCRs by Fluorescence Microscopy

            Samuel Génier, Jade Degrandmaison, Christine L. Lavoie, Louis Gendron, and Jean-Luc Parent

17. Combining RNAi and Immunofluorescence Approaches to Investigate Post-Endocytic Sorting of GPCRs into Multivesicular Bodies

            Xuezhi Li, Stéphanie Rosciglione, Andréanne Laniel, and Christine Lavoie

18. Super-Resolution Imaging of G Protein-Coupled Receptors Using Ground State Depletion Microscopy

            Fabiana A. Cateano-Crowley, Bryan Heit, and Stephen S.G. Ferguson

19. Analysis of Spatial Assembly of GPCRs Using Photoactivatable Dyes and Localization Microscopy

            Kim C. Jonas and Aylin C. Hanyaloglu

Part VI: Investigating In Vivo Functions of GPCRs

20. Optical Modulation of Metabotropic Glutamate Receptor Type 5 In Vivo Using a Photoactive Drug

            Marc López-Cano, Joan Font, Amadeu Llebaria, Víctor Fernández-Dueñas, and Francisco Ciruela

21. Imaging of Tissue-Specific and Temporal Activation of GPCR Signaling Using DREADD Knock-In Mice

            Dmitry Akhmedov, Nicholas S. Kirkby, Jane A. Mitchell, and Rebecca Berdeaux

22. Laser Doppler Flowmetry to Study the Regulation of Cerebral Blood Flow by G Protein-Coupled Receptors in Rodents

            Xavier Toussay, Mario Tiberi, and Baptiste Lacoste

23. Preparation of Pulmonary Artery Myocytes and Rings to Study Vasoactive GPCRs

            Martha Hinton, Anurag Singh Sikarwar, and Shyamala Dakshinamurti

Includes cutting-edge techniques

Provides step-by-step detail essential for reproducible results

Contains key implementation advice from the experts