Martins / Prazeres G Protein-Coupled Receptor Screening Assays

Methods and Protocols
2. Auflage 2021
ISBN: 978-1-0716-1221-7
Verlag: Springer US
Format: PDF
Kopierschutz: 1 - PDF Watermark

E-Book, Englisch, Band 2268, 327 Seiten, eBook

Reihe: Methods in Molecular Biology

This fully updated edition targets not only those assays directly involved in the discovery of GPCR-active compounds but also those involved in cell-based experiments designed to study physiological responses. Whether coming from academia or industry, or being an experienced researcher or a newcomer to the field, the reader will find accessible methods and protocols that cover the latest developments on receptor purification, molecular biology, recombinant engineering, and analytical techniques that enable the real time monitoring of the complex GPCR signaling cascade and identification of potential drug targets. 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.  Authoritative and up-to-date, G Protein-Coupled Receptor Screening Assays: Methods and Protocols, Second Edition aims to provide the tools necessary to contribute to the advancement of GPCR research and discovery and ultimately lead to the availability of innovative and more efficient drugs.

Zielgruppe

Professional/practitioner

Autoren/Hrsg.

Martins, Sofia Aires M.

Prazeres, Duarte Miguel F.

Inhaltsverzeichnis

Isolation of Lipid Rafts by the Detergent and Non-Detergent Based Methods for Localization of GPCRs with Immunoblotting and Laser Scanning Confocal Microscopy.- Detection of GPCR mRNA Expression in Primary Cells via qPCR, Microarrays, and RNA-Sequencing.- Construction of Recombinant Cell Lines for GPCR Expression.- Recombinant Expression and Purification of Cannabinoid Receptor CB 2 , a G Protein-Coupled Receptor.- Screening for Serotonin Receptor 4 Agonists Using GPCR-Based Sensor in Yeast.- Immobilization of Olfactory Receptors Carried by Nanosomes onto a Gold Sensor Surface.- Screening Methods for Cell-Free Synthesized GPCR/Nanoparticle Samples.- Fluorescence Anisotropy-Based Assay for Characterization of Ligand Binding Dynamics to GPCRs: The Case of Cy3B-Labelled Ligands Binding to MC 4 Receptors in Budded Baculoviruses.- Bioluminescence in G Protein-Coupled Receptors Drug Screening Using Nanoluciferase and Halo-Tag Technology.- Nanoluciferase-Based Complementation Assay to Detect GPCR-G Protein Interaction.- Imaging of Genetically-Encoded FRET-Based Biosensors to Detect GPCR Activity.- cAMP Biosensor Assay Using BacMam Expression System: Studying the Downstream Signaling of LH/hCG Receptor Activation.- FLIPR Calcium Mobilization Assays in GPCR Drug Discovery.- Live Cell Imaging and Optogenetics-Based Assays for GPCR Activity.- Split-Tobacco Etch Virus (Split-TEV) Method in G Protein-Coupled Receptor Interacting Proteins.- NanoLuc-Based Methods to Measure ß-Arrestin2 Recruitment to G Protein-Coupled Receptors.- Luciferase Complementation Approaches to Measure GPCR Signaling Kinetics and Bias.- Gradient Tracking by Yeast GPCRs in a Microfluidics Chamber.- Monitoring Intracellular Calcium in Response to GPCR Activation: Comparison Between Microtiter Plates and Microfluidic Assays.- Homology Modeling Using GPCRM Web Service.