MULTISCREEN™ STABLE CELL LINES
HUMAN RECOMBINANT GPR91 RECEPTOR
1 vial (2 x 106) frozen cells
Amsbio Cellbanker 2
Expression vector containing full-length human CB1 cDNA (GenBank Accession Number: NM_033050.3) with FLAG tag sequence at N-terminus.
Liquid nitrogen upon receiving
Propagation Medium: DMEM/F12, 10% FBS, 10 μg/mL puromycin
Stable in culture for minimum of two months
Background: GPR91, also known as SUCNR1, is a G Protein-Coupled Receptor with 339 amino acids. It has been characterized as a receptor for Succinate, a citric acid cycle intermediate. Succinate plays a key role in energy metabolism. Local interstitial accumulation of Succinate has recently been reported to serve as an indicator of ischemic or diabetic organ damage in the brain, liver, and kidney. In diabetes patients, the accumulation of Succinate is detectable in the plasma, and more significantly in the renal tubular fluid and urine. It is therefore considered a potential new biomarker of local tissue damage. It has also been shown that Succinate increases blood pressure in animals. The Succinate-induced hypertensive effect involves the renin-angiotensin system that is shown to be absent in GPR91-deficient mice. There is a possible role for GPR91 in renovascular hypertension, a disease closely linked to atherosclerosis, diabetes and renal failure. In a recombinant system overexpressing GPR91, Succinate was shown to not only stimulate calcium mobilization and inositol phosphate (IP) accumulation through the stimulation of Gαq pathway but also to activate the Erk1/2 MAPK pathway and inhibit forskolin-stimulated cAMP accumulation through Gαi pathway.
Application: Functional assays
Figure 1. Dose-dependent stimulation of calcium flux upon treatment with ligand, measured with MultiscreenTM Calcium 1.0 No Wash Assay Kit (Multispan MSCA01). Figure 2. Dose-dependent inhibition of forskolin-stimulated intracellular cAMP accumulation upon treatment with ligand, measured with MultiscreenTM TR-FRET cAMP 1.0 No Wash Assay Kit (Multispan MSCM01). Figure 3. Dose response of intracellular IP1 accumulation upon treatment with ligand, measured with IP-one Tb kit (Cisbio 62IPAPEC). Figure 4. Receptor expression on cell surface measured by flow cytometry (FACS) using an anti-FLAG antibody. Thin line: parental cells; thick line: receptor-expressing cells.
He, W., Miao, F. J., Lin, D. C., Schwandner, R. T., Wang, Z., Gao, J., Chen, J. L., Tian, H. and Ling, L. (2004) Citric acid cycle intermediates as ligands for orphan G-protein-coupled receptors. Nature, 429: 188-193.
Peti-Peterdi, J. (2010) “High glucose and renin release: the role of succinate and GPR91.” Kidney International, 78(12):1214-7