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
eBook ISBN:
9780123918734
Print ISBN:
9780123918628

About the serial-volume-editor

P. Michael Conn

P. Michael Conn is the Senior Vice President for Research and Associate Provost, Texas Tech Health Sciences Center. He is The Robert C. Kimbrough, Professor of Internal Medicine and Cell Biology/Biochemistry. He was previously Director of Research Advocacy and Professor of Physiology and Pharmacology, Cell Biology and Development and Obstetrics and Gynecology at Oregon Health and Science University and Senior Scientist of the Oregon National Primate Research Center (ONPRC). He served for twelve years as Special Assistant to the President and Associate Director of the ONPRC. After receiving a B.S. degree and teaching certification from the University of Michigan (1971), a M.S. from North Carolina State University (1973), and a Ph.D. degree from Baylor College of Medicine (1976), Conn did a fellowship at the NIH, then joined the faculty in the Department of Pharmacology, Duke University Medical Center where he was promoted to Associate Professor in 1982. In 1984, he became Professor and Head of Pharmacology at the University of Iowa College of Medicine, a position he held for eleven years. Conn is known for his research in the area of the cellular and molecular basis of action of gonadotropin releasing hormone action in the pituitary and therapeutic approaches that restore misfolded proteins to function. His work has led to drugs that have benefitted humans and animals. Most recently, he has identified a new class of drugs, pharmacoperones, which act by regulating the intracellular trafficking of receptors, enzymes and ion channels. He has authored or co-authored over 350 publications in this area and written or edited over 200 books, including texts in neurosciences, molecular biology and endocrinology. Conn has served as the editor of many professional journals and book series (Endocrinology, Journal of Clinical Endocrinology and Metabolism, Endocrine, Methods, Progress in Molecular Biology and Translational Science and Contemporary Endocrinology). Conn serve

Affiliations and Expertise

Texas Tech University Health Sciences Center, Lubbock, USA