A green-fluorescent-protein-based assay for the characterization of G-protein-coupled receptors
| Autor: | Derk Görich, Thomas Roeder, Dörte Heyden, Michael Gewecke |
|---|---|
| Rok vydání: | 2004 |
| Předmět: |
Biophysics
Cell Biology Biology Ligands Transfection Ligand (biochemistry) Biochemistry Embryonic stem cell Receptors G-Protein-Coupled Green fluorescent protein Cell biology chemistry.chemical_compound chemistry Genes Reporter COS Cells Expression cloning Animals Humans Receptors Histamine H2 Receptor Molecular Biology Histamine Signal Transduction G protein-coupled receptor |
| Zdroj: | Analytical Biochemistry. 332:38-45 |
| ISSN: | 0003-2697 |
| DOI: | 10.1016/j.ab.2004.05.023 |
| Popis: | A simple and sensitive system to monitor activation and pharmacology of G-protein-coupled receptors in mammalian cells is described. It is based on cAMP-responsible-element-regulated expression of green fluorescent protein (GFP). Cotransfection with appropriate G-protein-coupled receptors and subsequent activation with agonists induces expression of GFP in a dose-dependent manner. This system is suited for the analysis of most G-protein-coupled receptors, including those that are coupled to Gs, Gi, and Gq. It can replace reporter systems for G-protein-coupled receptors currently in use. Time-consuming and labor-intensive analysis is avoided because it is a noninvasive system, which allows multiple reprobing without disturbing the cells. In addition, adaptation to high-throughput approaches is possible. Together with human embryonic kidney cells, it is a zero-background expression system, making it ideally suited to the pharmacological characterization of cloned receptors, to expression cloning experiments, and to the identification of the natural ligand of orphan G-protein-coupled receptors. |
| Databáze: | OpenAIRE |
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