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G Protein-Coupled Receptors

Signaling, Trafficking and Regulation

1st Edition - February 26, 2016

Editor: Arun K. Shukla

Hardback ISBN:

9 7 8 - 0 - 1 2 - 8 0 3 5 9 5 - 5

eBook ISBN:

9 7 8 - 0 - 1 2 - 8 0 3 6 1 2 - 9

G-Protein-Coupled Receptors: Signaling, Trafficking, and Regulation, a new volume in the Methods in Cell Biology series continues the legacy of this premier serial with quality… Read more

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G-Protein-Coupled Receptors: Signaling, Trafficking, and Regulation, a new volume in the Methods in Cell Biology series continues the legacy of this premier serial with quality chapters authored by leaders in the field. This volume covers research methods in G-Protein-Coupled Receptors, and includes sections on such topics signaling, trafficking and regulation.

Series Editors
Preface
Section I: Trafficking, Localization and Imaging of GPCRs
- Chapter 1. Localization and signaling of GPCRs in lipid rafts
  - Introduction
  - 1. Localization of GPCRs in Lipid Rafts
  - 2. GPCR Signaling in Lipid Rafts
- Chapter 2. Imaging GPCRs trafficking and signaling with total internal reflection fluorescence microscopy in cultured neurons
  - 1. Image Acquisition
  - 2. Image Analysis
  - 3. Final Comments
- Chapter 3. Trafficking of ciliary G protein-coupled receptors
  - Introduction
  - 1. Rhodopsin as a Ciliary GPCR
  - 2. Ciliary GPCRs in Other Neurons
  - 3. Olfactory Sensory Neuron Ciliary GPCRs
  - 4. Dynamic Trafficking of Ciliary GPCRs
  - Conclusions and Future Directions
- Chapter 4. Single-molecule resolution of G protein-coupled receptor (GPCR) complexes
  - Introduction
  - 1. Imaging Methodologies to Study Single GPCR Molecules
  - 2. Methods for Photoactivated Dye-Localization Microscopy
  - Conclusions and Perspectives
- Chapter 5. Quantification of the mRNA expression of G protein-coupled receptors in human adipose tissue
  - Introduction
  - 1. Materials
  - 2. Methods
  - Notes
Section II: Signaling and Regulation of GPCRs
- Chapter 6. Studying the regulation of endosomal cAMP production in GPCR signaling
  - Introduction
  - 1. Materials
  - 2. Experimental Procedures
  - 3. Discussion
- Chapter 7. Olfactory receptor signaling
  - Introduction
  - 1. Specificity
  - 2. Dimerization
  - 3. Perception
  - 4. Olfactory Transduction
  - 5. Mechanisms of Odorant Adaptation
  - 6. Perspectives
- Chapter 8. Assessing Smoothened-mediated Hedgehog signaling in zebrafish
  - Introduction
  - 1. Analysis of Smoothened-mediated Hh Signaling in Zebrafish
  - Summary
- Chapter 9. GPCRs and actin–cytoskeleton dynamics
  - Introduction
  - 1. GPCRs and G Proteins
  - 2. Actin–cytoskeleton Dynamics
  - 3. GPCR/G Protein Signaling and Actin Polymerization
  - 4. Cellular Processes Controlled by Actin–cytoskeleton Rearrangements Induced by GPCRs
  - 5. The Cross Talk of GPCR Signaling with the Homeostatic Hippo and TGF-β Signaling Pathways
  - 6. Protocols to Evaluate Actin–cytoskeleton Rearrangements
  - 7. Method to Evaluate the Actin–Cytoskeleton Dynamics Modulated by GPCR Signaling in Hepatic Cells
  - 8. Protocols for IF Assays
  - Conclusions
Section III: Cellular Assays for GPCRs
- Chapter 10. GPCR-radioligand binding assays
  - 1. Types of Binding Assays
  - 2. Components of Binding Assays—Considerations for GPCR Binding Assay Design
  - Abbreviations
- Chapter 11. Tracking GPCR biosynthesis and degradation using a nonradioactive pulse chase methodology
  - Introduction
  - 1. Materials Required
  - 2. Basic Methodology
  - 3. General Workflow of Experiments
  - 4. Results
  - 5. Discussion
- Chapter 12. Tango assay for ligand-induced GPCR–β-arrestin2 interaction: Application in drug discovery
  - 1. Background and Rationale
  - 2. Various β-Arrestin Recruitment Assays
  - 3. Principle of Tango Assay
  - 4. Reagents and Methods
  - 5. Data Analysis
  - 6. Results and Data Interpretation
  - 7. Summary and Future Perspectives
- Chapter 13. Resonance Energy Transfer-Based Approaches to Study GPCRs
  - Introduction
  - 1. RET Approaches
  - 2. Ligand Binding
  - 3. Ligand-Induced Conformational Changes
  - 4. GPCR–G Protein Interaction
  - 5. Second Messenger Production
  - 6. β-Arrestin Recruitment
  - 7. Receptor Internalization
  - 8. Receptor Oligomerization
  - 9. Summary and Outlook
- Chapter 14. Quantitative analysis of G-protein-coupled receptor internalization using DnaE intein-based assay
  - Introduction
  - 1. Materials and Methods
  - 2. Discussion
  - Summary
- Chapter 15. Cellular and subcellular context determine outputs from signaling biosensors
  - Introduction and Rationale
  - 1. Materials and Instrumentation Required
  - 2. Basic Methodology
  - 3. Experimental Strategies
  - 4. Discussion
Section IV: Structural and Computational Investigation of GPCRs
- Chapter 16. Protease-activated receptors (PARs) in cancer: Novel biased signaling and targets for therapy
  - Introduction
  - 1. Novel Signaling of PARs Endowing Critical PH-Domain-Binding Motifs
  - 2. PARs and Human Placenta
  - 3. Future Directions
- Chapter 17. Computational methods for studying G protein-coupled receptors (GPCRs)
  - Introduction
  - 1. Prediction of Receptor Structure
  - 2. Studies of Ligand–Receptor Interactions
  - 3. Biased Signaling
  - 4. Allosterism
  - 5. Dimerization
- Chapter 18. Comparing Class A GPCRs to bitter taste receptors: Structural motifs, ligand interactions and agonist-to-antagonist ratios
  - Introduction
  - 1. Sequence Is Less Conserved Than Structure: Sequence Alignments and GPCR Signature Motifs
  - 2. Ligand–Receptor Interactions in TAS2Rs Compared to Class A GPCRs
  - Conclusion and Outlook
- Chapter 19. What can simulations tell us about GPCRs: Integrating the scales
  - Introduction
  - 1. Multiscale Simulations: Accuracy, Advantages, and Limitations
  - 2. Analyzing the Dynamics of GPCR–Ligand Interactions
  - 3. Can Current Simulation Techniques Allow Us to Study GPCR Activation?
  - 4. The Dynamic Interplay between the Membrane and Receptors
  - 5. GPCR Organization in the Cell Membrane
  - Conclusions
- Volumes in Series
- Index