title: High-throughput screening of interactions between G protein-coupled receptors and ligands using confocal optics microscopy.
abstract: Interactions of extracellular ligands with proteins in the cellular plasma membrane are the starting point for various intracellular signaling cascades. In the pharmaceutical industry, particular attention has been paid to G protein- coupled receptors (GPCRs), which are involved in various disease processes. In so-called high-throughput screening (HTS) campaigns, large medicinal chemistry compound libraries were searched for bioactive molecules that would either induce or inhibit the activity of a specific disease-relevant GPCR. In the respective drug discovery assays, the test compound typically competes with the physiological ligand for a binding site on the receptor. The transmembrane receptor is prepared in the form of membrane fragments or, as described here, in so-called virus-like particles (VLiPs). As hundreds of thousands of test compounds must be analyzed, there is a strict need for low volume binding assays to save the expensive bioreagents, and to reduce the consumption of the test compounds. In this chapter, we describe the application of confocal optics microscopy to measure GPCR ligand interactions in low microliter assay volumes.
authors.affiliation: Department of Biophysics, University of Ulm, Ulm, Germany.
authors.forename: Lenka
authors.initials: L
authors.lastname: Zemanová
authors.forename: Andreas
authors.initials: A
authors.lastname: Schenk
authors.forename: Martin J
authors.initials: MJ
authors.lastname: Valler
authors.forename: G Ulrich
authors.initials: GU
authors.lastname: Nienhaus
authors.forename: Ralf
authors.initials: R
authors.lastname: Heilker
mesh_terms: D000818:Animals; D001331:Automation; D004353:Drug Evaluation, Preclinical; D005454:Fluorescence Polarization; D066298:In Vitro Techniques; D008024:Ligands; D018613:Microscopy, Confocal / Q000295:instrumentation / Q000379:methods*; D008904:Miniaturization; D015394:Molecular Structure; D011485:Protein Binding; D043562:Receptors, G-Protein-Coupled / Q000737:chemistry* / Q000378:metabolism*; D011994:Recombinant Proteins / Q000737:chemistry / Q000378:metabolism; D014771:Virion
chemical_list: D008024:Ligands; D043562:Receptors, G-Protein-Coupled; D011994:Recombinant Proteins