MULTISCREEN™ β-ARRESTIN STABLE CELL LINES
HUMAN RECOMBINANT GPR4 RECEPTOR
1 vial (2 x 106) frozen cells
Cellbanker 2 (Amsbio 11891)
Expression vector containing full-length human GPR4 cDNA (GenBank Accession Number NM_005282.1) with FLAG tag sequence at N-terminus and ARRB2 cDNA (GenBank Accession Number NM_004313.3)
Liquid nitrogen upon receiving
Propagation Medium: DMEM, 10% FBS, 1 μg/mL puromycin, 250 μg/mL hygromycin
Stable for a minimum of 2 months in continuous culture
Background: The GPR4 is a family of proton-sensing G protein-coupled receptors (GPCRs) and has recently been identified as novel pH sensors. GPR4 sense extracellular protons through histidine residues of the receptors and are coupled to G-proteins to stimulate intracellular signaling pathways. This receptor is expressed in vascular endothelial and smooth muscle cells, as well in a wide range of tissues such as the lung, kidney, heart, and liver. GPR4, upon activation by acidic pH stimulates the Gs/cyclic adenosine monophosphate (cAMP) signaling pathway in endothelial cells and regulates micro vessel growth.
Application: Functional assays
Figure 1. Dose-dependent stimulation from arrestin recruitment upon treatment with pH, measured with MultiscreenTM β-Arrestin Assay Kit (Multispan MSBAK01). Figure 2. Dose- dependent increase of intracellular cAMP level upon treatment with ligand, measured with MultiscreenTM TR-FRET cAMP 1.0 No Wash Assay Kit (Multispan MSCM01). Figure 3. Receptor expression on cell surface measured by flow cytometry (FACS) using an anti- FLAG antibody. Thin line: parental cells; thick line: receptor-expressing cells.
Ludwig, M.-G., Vanek, M., Guerini, D., Gasser, J. A., Jones, C. E., Junker, U., Hofstetter, H., Wolf, R. M., Seuwen, K. Proton-sensing G-protein-coupled receptors. Nature 425: 93- 98, 2003.
Chen A, Dong L, Leffler NR, Asch AS, Witte ON, et al. (2011) Activation of GPR4 by Acidosis Increases Endothelial Cell Adhesion through the cAMP/Epac Pathway. PLOS ONE 6(11): e27586. doi: 10.1371/journal.pone.0027586
Yang LV, Radu CG, Roy M, Lee McLaughlin J, et al. (2007) Vascular abnormalities in mice deficient for the G protein- coupled receptor GPR4 that functions as a pH sensor. Mol Cell Biol 27: 1334–1347