G Protein-Coupled Receptors in Drug Discovery

This detailed volume provides an overview of recent techniques employed in the field of G protein-coupled receptors (GPCRs) to screen for new drugs and to derive information about their receptor structure, dynamics, and function for the purpose of developing improved therapeutics. Owing to remarkable recent advances in the structural, biophysical and biochemical analyses of these receptors, as well as a growing body of evidence hinting at the possible relevance of allosteric modulators, biased agonists and oligomer-selective ligands as improved therapeutic agents, drug discovery for GPCRs has recently taken a completely new direction. For this book, expert contributors have shared their protocols and views on the impact of these methodologies on modern drug discovery. Written for the highly successful Methods in Molecular Biology series, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols and tips on troubleshooting and avoiding known pitfalls.

Practical and fully updated, G Protein-Coupled Receptors in Drug Discovery: Methods and Protocols, Second Edition serves as an ideal guide for a diverse audience from structural and molecular biologists to pharmacologists and drug designers who wish to explore this extensive class of key drug targets.

Includes cutting-edge methods and protocols on GPCRs in drug discovery Provides step-by-step detail essential for reproducible results Contains key notes and implementation advice from the experts

Inhalt

Purification of Stabilized GPCRs for Structural and Biophysical Analyses.- Purification and Crystallization of a Thermostabilized Agonist-Bound Conformation of the Human Adenosine A 2A Receptor.- 2D Projection Analysis of GPCR Complexes by Negative Stain Electron Microscopy.- Nuts and Bolts of CF 3 and CH 3 NMR Towards the Understanding of Conformational Exchange of GPCRs.- Single-Molecule Fluorescence Microscopy for the Analysis of Fast Receptor Dynamics.- Quantitative Multi-Color Detection Strategies for Bioorthogonally-Labeled GPCRs.- Approaches to Characterize and Quantify Oligomerization of GPCRs.- Monitoring G Protein Activation in Cells with BRET.- Use of Fluorescence Indicators in Receptor Ligands.- Detection and Quantification of Intracellular Signalling Using FRET-Based Biosensors and High Content Imaging.- The Measurement of Receptor Signaling Bias.- Approaches to Assess Functional Selectivity in GPCRs: Evaluating G Protein Signaling in an Endogenous Environment.- Bioluminescence Resonance Energy Transfer Approaches to Discover Bias in GPCR Signaling.- Virus-Mediated Expression of DREADDs for In Vivo Metabolic Studies.- High-Throughput Screening for Allosteric Modulators of GPCRs.- Radioligand Binding Assay for an Exon 11-Associated Mu Opioid Receptor Target.- Docking and Virtual Screening Strategies for GPCR Drug Discovery.- The Dynamic Process of Drug-GPCR Binding at either Orthosteric or Allosteric Sites Evaluated by Metadynamics.- Experiment-Guided Molecular Modeling of Protein-Protein Complexes Involving GPCRs.- Interaction Fingerprints and Their Applications to Identify Hot Spots.