Prazeres D.M.F., Martins S.A.M. (Eds.) G Protein-Coupled Receptor Screening Assays. Methods and Protocols

Humana Press, 2015 — 296p. — (Methods in Molecular Biology 1272) — ISBN: 978-1-4939-2336-6 (eBook), 978-1-4939-2335-9 (Hardcover).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.G protein-Coupled Receptors: An Overview of Signaling Mechanisms and Screening Assays
Time-Resolved FRET Strategy to Screen GPCR Ligand Library
Homogeneous Fluorescence Anisotropy-Based Assay for Characterization of Ligand Binding Dynamics to GPCRs in Budded Baculoviruses: The Case of Cy3B-NDP-α-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
Multidimensional GPCR Profiling and Screening Using Impedance-Based Label-Free and Real-Time Assay
Label-Free Functional Selectivity Assays
Measurement of Surface-Mediated Ca²⁺ 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