G Protein-Coupled Receptors in Drug Discovery, Softcover reprint of the original 1st ed. 2009
Methods in Molecular Biology Series, Vol. 552

The G protein-coupled receptors (GPCRs) and associated peripheral G proteins underpin a multitude of physiological processes. The GPCRs represent one of the largest superfamilies in the human genome and are a significant target for bioactive and drug discovery programs. It is estimated that greater than 50% of all drugs, including those in development, currently target GPCRs. Many of the characterized GPCRs have known ligands; however, approximately 20% of GPCRs are described as orphan GPCRs, apparent GPCRs that share the generic high-level structure charact- istic of GPCRs but whose endogenous ligand is not known. Therefore, it is expected that the field of GPCR drug discovery and development will greatly expand in the coming years with emphasis on new generations of drugs against GPCRs with unique therapeuticuseswhichmayincludedrugssuchasallostericregulators,inverseagonists, and identification of orphan GPCR ligands. AswelearnmoreaboutthemolecularsignalingcascadesfollowingGPCRactivation, we acquire a better appreciation of the complexity of cell signaling and as a result, also acquire a vast array ofnew molecularmethods toinvestigate these andother processes. Thegeneralaimofthisbookistoprovideresearcherswitharangeofprotocolsthatmay be useful in their GPCR drug discovery programs. It is also the basis for the devel- ment of future assays in this field. Therefore, the range of topics covered and the appropriate methodological approaches in GPCR drug discovery are reflected in this book. Itisinterestingtonotethatfuturedirectionsindrugdiscoverywillrequireinput and collaboration from a plethora of fields of research. As such, this book will likely be of interest to scientists involved in such fields as molecular biology, pharmacology, biochemistry, cellular signaling, andbio-nanotechnology.

New Insights into GPCR Function: Implications for HTS.- Screening Technologies for G Protein-Coupled Receptors: From HTS to uHTS.- GPCR Signaling: Understanding the Pathway to Successful Drug Discovery.- An Overview on GPCRs and Drug Discovery: Structure-Based Drug Design and Structural Biology on GPCRs.- Understanding the Ligand–Receptor–G Protein Ternary Complex for GPCR Drug Discovery.- Assay Data Quality Assessment.- Homology Modeling of GPCRs.- GPCR Expression Using Baculovirus-Infected Sf9 Cells.- Radioligand Binding Assays: Application of [125I]Angiotensin II Receptor Binding.- [35S]GTP?S Binding in G Protein-Coupled Receptor Assays.- A Time-Resolved Fluorescent Lanthanide (Eu)-GTP Binding Assay for Chemokine Receptors as Targets in Drug Discovery.- Use of the DiscoveRx Hithunter cAMPII Assay for Direct Measurement of cAMP in Gs and Gi GPCRs.- Use of Aequorin for G protein-Coupled Receptor Hit Identification and Compound Profiling.- BacMam: Versatile Gene Delivery Technology for GPCR Assays.- Yeast Assays for G Protein-Coupled Receptors.- GPCR Microspot Assays on Solid Substrates.- Resonant Waveguide Grating Biosensor for Whole-Cell GPCR Assays.- FRET-Based Measurement of GPCR Conformational Changes.- FLIPR® Assays of Intracellular Calcium in GPCR Drug Discovery.- Use of Fluorescence Indicators in Receptor Ligands.- Application of Fluorescence Resonance Energy Transfer Techniques to Establish Ligand–Receptor Orientation.- Detection of GPCR/?-Arrestin Interactions in Live Cells Using Bioluminescence Resonance Energy Transfer Technology.- Using Reporter Gene Technologies to Detect Changes in cAMP as a Result of GPCR Activation.- A Quantum Dot-Labeled Ligand–Receptor Binding Assay for G Protein-Coupled Receptors Contained in Minimally Purified MembraneNanopatches.- Xenopus Oocyte Electrophysiology in GPCR Drug Discovery.- Immunoprecipitation and Phosphorylation of G Protein-Coupled Receptors.

Contains several comprehensive reviews of GPCR screening technologies by authoritative leaders Provides an easily accessible reference volume for cell-based GPCR screening written by leading researchers in the field Covers a wide range of assay formats and systems Emphasizes technologies applicable to high throughput screening and research-based, high information content assays Presents a diverse approach to experimental design, covering model biological systems, screening modalities, detection technologies, and approaches to data analysis Comprehensive guide to developing, running, and analyzing cell-based assays in high-throughput screening Includes supplementary material: sn.pub/extras