Description

This new volume of Methods in Enzymology continues the legacy of this premier serial by containing quality chapters authored by leaders in the field. This volume covers G protein coupled receptors and includes chapters on such topics as G protein-coupled receptor trafficking motifs, structure-based virtual screening, and automation-friendly high throughput assays for identification of pharmacoperone drugs.

Key Features

  • Continues the legacy of this premier serial with quality chapters authored by leaders in the field
  • Covers G protein coupled receptors
  • Contains chapters on such topics as G protein-coupled receptor trafficking motifs, structure-based virtual screening, and automation-friendly high-throughput assays for identifying pharmacoperone drugs

Readership

Biochemists, biophysicists, molecular biologists, analytical chemists, and physiologists

Table of Contents

Series Page

Contributors

Preface

Methods in Enzymology

Chapter One. Therapeutic Rescue of Misfolded/Mistrafficked Mutants

1. Introduction

2. Choosing Pharmacoperone Model Systems

3. Selection of Endpoint Measures

4. Assay Automation

5. Data Analysis

6. Hit Follow-Up Experiments

7. Conclusions

8. Acknowledgments

References

Chapter Two. Trafficking of the Follitropin Receptor

1. Introduction

2. Outward Trafficking Defective FSHR Mutants. Studying Plasma Membrane Expression of the FSHR

3. Studying Oligomerization of Intracellular and Cell Surface-Expressed FSHRs

4. Phosphorylation, Internalization, and Recycling of the FSHR (Downward Trafficking)

5. Acknowledgments

References

Chapter Three. Single-Molecule Imaging Technique to Study the Dynamic Regulation of GPCR Function at the Plasma Membrane

1. Introduction

2. Labeling of GPCRs and Sample Preparation for SPT

3. Single-Molecule Imaging

4. Data Analysis

5. Concluding Remarks

6. Acknowledgments

References

Chapter Four. GPCR Oligomerization and Receptor Trafficking

1. Introduction

2. GPCR Expression Using the Flp-In™ T-Rex™ System

3. Detecting GPCR Internalization by Fluorescent Microscopy: The Orexin OX1 Receptor

4. SNAP–CLIP Tagging

5. Detecting GPCR Oligomerization by other Resonance Energy Transfer Techniques

6. Biotinylation Studies

7. ER Trapping, Pharmacological Chaperones, and the Use of Engineered Synthetic Ligands

8. Acknowledgments

References

Chapter Five. β-Arrestins and G Protein-Coupled Receptor Trafficking

1. Introduction

2. Arrestin Expression

3. Assays to Measure GPCR Trafficking

4. Evaluating the Role of β-Arrestins in GPCR Traffick

Details

No. of pages:
480
Language:
English
Copyright:
© 2013
Published:
Imprint:
Academic Press
Print ISBN:
9780123918628
Electronic ISBN:
9780123918734