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 post-translation modification of GPCR in relationship to biased agonism, structure-based virtual screening, and GPCR oligomerization in the brain.

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 post-translation modification of GPCR in relationship to biased agonism, structure-based virtual screening, and GPCR oligomerization in the brain

Readership

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

Table of Contents

Series Page

Contributors

Preface

Methods in Enzymology

Chapter One. Expression of GPCRs in Pichia pastoris for Structural Studies

1 Introduction

2 Generation of Expression Clones

3 Screening Transformants

4 Characterization and Optimization of Initial Target Receptor Production

5 Large-Scale Production

6 Prospects for Large-Scale GPCR Production in P. pastoris

References

Chapter Two. Conformational Ensemble View of G Protein-Coupled Receptors and the Effect of Mutations and Ligand Binding

1 Introduction

2 Overview of the Conformational Ensemble Prediction

3 Generating Starting Structures Based on Templates

4 BiHelix: TM Bundle Conformational Sampling of Helix Rotation Angles

5 SuperBiHelix TM Bundle Sampling of All Helix Orientation Angles

6 Effect of Mutations on the WT Conformational Ensemble

7 Ensemble Docking of Ligands to WT or Mutant Receptors

8 Summary

References

Chapter Three. Structural Evolution of G-Protein-Coupled Receptors

Abbreviations

1 Introduction

2 The Puzzle of GPCR Evolution

3 The Evolution of Class A GPCRs Viewed by MDS

4 Evolutionary Trends

5 Structural Evolution of GPCRs

6 Summary

References

Chapter Four. Directed Evolution of G-Protein-Coupled Receptors for High Functional Expression and Detergent Stability

1 Introduction

2 Methods

References

Chapter Five. The Role of Hydrophobic Amino Acids in the Structure and Function of the Rhodopsin Family of G Protein-Coupled Receptors

1 Introduction

2 The Structure of GPCRs

3 Sequence Analyses of the 7TM Segments of the Rhodopsin Family of GPCRs

4 Importance of the Highly Conserved Hydrophobic Amino Acid at Position 3.40 in the Process of Agonist-Induced Receptor Activation

5 The Hydro

Details

No. of pages:
440
Language:
English
Copyright:
© 2013
Published:
Imprint:
Academic Press
Electronic ISBN:
9780123918727
Print ISBN:
9780123918611

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