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This volume explores the considerable efforts that have been directed towards the development of G Protein-Coupled Receptors (GPCR) screening assays in order to disclose GPCR acting compounds, elucidate signaling mechanisms or evaluate compound's efficacy. New discoveries in the field, along with the widely recognized need for better and safer pharmaceutical drugs constitute the main motivation for this book. Readers, both beginners and experienced researchers, will receive an updated overview of not only the established, but also the innovative technologies that promise to advance GPCR drug research. This book is organized into two major parts: the introductory part discusses the necessary foundations for the understanding of GPCR action and the rationale behind the design of the available screening assays; and part two provides detailed protocols for different screening approaches. Written in the highly successful Methods in Molecular Biology series format, the chapters include the kind of detailed description and implementation advice that is crucial for getting optimal results in the laboratory.
Practical and innovative, G Protein-Coupled Receptor Screening Assays: Methods and Protocols reaches out to everyone involved in the discovery of GPCR-active drugs, and provides a transversal overview of the different levels of GPCR signaling addressable in the different screening strategies and presents practical examples of how current assay technologies are contributing to new paradigms in GPCR drug research.
List of contents
G-Protein Coupled Receptors: An Overview of Signaling Mechanisms and Screening Assays.- Time-Resolved FRET Strategy to Screen GPCR Ligands Library.- Homogeneous Fluorescence Anisotropy-Based Assay for Characterization of Ligand Binding Dynamics to GPCRs in Budded Baculoviruses: The Case of Cy3B-NDP-a-MSH Binding to MC4 Receptors.- Construction of Recombinant HEK293 Cell Lines for the Expression of the Neurotensin Receptor NTSR1.- cAMP Assay for GPCR Ligand Characterization: Application of BacMam Expression System.- Ca2+ Mobilization Assays in GPCR Drug Discovery.- Using Constitutive Activity to Define Appropriate High Throughput Screening Assays for Orphan G Protein-Coupled Receptors.- Monitoring G-Protein Coupled Receptor Activation Using the Protein Fragment Complementation Technique Split TEV.- Quantifying GPCR Internalization: A Focus on the Kisspeptin Receptor.- GPCR Oligomerization Analysis by Means of BRET and dFRAP.- Use of ImageJ to Recover Information from Individual Cells in a G-Protein Coupled Receptor Assay.- Methods to Immobilize GPCR on the Surface of SPR Sensors.- Olfactory Receptor Screening Assay Using Nanovesicle-Immobilized Carbon Nanotube Transistor.- Label-Free Biosensor Assays in GPCR Screening.- Multi-Dimensional GPCR Profiling and Screening Using Impedance-Based Label-Free and Real-Time Assay.- Label-Free Functional Selectivity Assays.- Measurement of Surface-Mediated Ca++ Transients on the Single-Cell Level in a Microfluidic Lab-on-a-chip Environment.- Cell-Based Assays and Animal Models for GPCR Drug Screening.- Computer-Aided Design of GPCR Ligands.
Summary
This volume explores the considerable efforts that have been directed towards the development of G Protein-Coupled Receptors (GPCR) screening assays in order to disclose GPCR acting compounds, elucidate signaling mechanisms or evaluate compound’s efficacy. New discoveries in the field, along with the widely recognized need for better and safer pharmaceutical drugs constitute the main motivation for this book. Readers, both beginners and experienced researchers, will receive an updated overview of not only the established, but also the innovative technologies that promise to advance GPCR drug research. This book is organized into two major parts: the introductory part discusses the necessary foundations for the understanding of GPCR action and the rationale behind the design of the available screening assays; and part two provides detailed protocols for different screening approaches. Written in the highly successful Methods in Molecular Biology series format, the chapters include the kind of detailed description and implementation advice that is crucial for getting optimal results in the laboratory.
Practical and innovative, G Protein-Coupled Receptor Screening Assays: Methods and Protocols reaches out to everyone involved in the discovery of GPCR-active drugs, and provides a transversal overview of the different levels of GPCR signaling addressable in the different screening strategies and presents practical examples of how current assay technologies are contributing to new paradigms in GPCR drug research.
