G Protein Coupled Receptors Structure 1st Edition by Michael Conn 9780123918611 0123918618

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ISBN 10: 0123918618
ISBN 13: 9780123918611
Author: P. Michael Conn

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.

• 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

G Protein Coupled Receptors Structure 1st Table of contents:

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

1. Introduction
2. Generation of Expression Clones
   • P. pastoris strains
   • P. pastoris expression vectors
   • High efficiency electroporation
3. Screening Transformants
   • Single picking from initial transformation plates
   • Assessing multiple expression cassette insertions
   • Direct screening for GPCR production
4. Characterization and Optimization
   • Small-scale expression (20mL)
   • Membrane preparation
   • Optimizing functional receptor expression
5. Large-Scale Production
   • 6x 1L liquid shaker culture
   • Fermentor-scale production
   • Large-scale membrane preparation
6. Prospects for Large-Scale GPCR Production

Chapter Two: Conformational Ensemble View of GPCRs and the Effect of Mutations

1. Introduction
2. Overview of Conformational Ensemble Prediction
3. Generating Starting Structures
4. BiHelix: Helix Rotation Sampling
5. SuperBiHelix: Helix Orientation Sampling
6. Effect of Mutations
7. Ensemble Docking of Ligands
   • Agonists and antagonists to A3 adenosine receptor
   • Ligand docking to CCR5 receptor mutants
8. Summary

Chapter Three: Structural Evolution of GPCRs

1. Introduction
2. The Puzzle of GPCR Evolution
3. Evolution of Class A GPCRs
   • Sequence preparation
   • MDS analysis
   • Sequence space analysis
4. Evolutionary Trends
   • Hallmark residues
   • Mono vs. polyphyletic groups
   • Evolutionary drift of subfamilies
5. Structural Evolution of GPCRs
   • TM2 proline pattern
   • TM2 and TM5 proline mutations
6. Summary

Chapter Four: Directed Evolution of GPCRs for High Expression and Stability

1. Introduction
   • GPCR structures
   • Engineering approaches
2. Methods
   • Genetic diversity generation
   • Expression and selection
   • Characterization of selected variants

Chapter Five: Hydrophobic Amino Acids in Rhodopsin Family GPCRs

1. Introduction
2. GPCR Structure
3. Sequence Analyses of 7TM Segments
4. Functional Role of Hydrophobic Residues
5. Transmembrane Aqueous Channel
6. GPCR Oligomerization
7. Conclusions

Chapter Six: Structure of β-Adrenergic Receptors

1. Introduction
2. Engineering and Expression
3. Purification and Crystallization
4. Structural Insights

Chapter Seven: Advances in Characterizing Ligand-Induced GPCR Activation

1. Introduction
2. Experimental Principles
3. Detailed Experimental Procedures

Chapter Eight: Crystallogenesis of A2A Receptor-T4 Lysozyme Fusion

1. Introduction
2. Fusion Receptor Design
3. Purification and Crystallization
4. Data Collection

Chapter Nine: Probing GPCR Structure: Adenosine and P2Y Receptors

1. Introduction
2. Extracellular Nucleoside Receptors
3. Extracellular Nucleotide Receptors
4. Neoceptors Approach

Chapter Ten: Photoaffinity Labeling of GPCR Binding Sites

1. Introduction
2. VPAC1 Receptor as a Model
3. Photoaffinity Labeling Methods

Chapter Eleven: Expression of GPCRs in C. elegans

1. Introduction
2. Transgenic Expression
3. Detection and Purification
4. Functional Assays

Chapter Twelve: Structural Analysis of mGluR Intracellular C-Termini

1. Introduction
2. mGluR-CT Synthesis
3. Structural Characterization

Chapter Thirteen: Unnatural Amino Acid Mutagenesis of GPCRs

1. Introduction
2. Site-Specific Mutagenesis
3. Labeling Strategies

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Michael Conn,Protein Coupled,Receptors