Calcium Flux Assay

HTS206C - ChemiSCREEN™ Human Recombinant MrgD Mas-Related Receptor Calcium-Optimized Stable Cell Line

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    The Mas-related genes (Mrgs), also known as sensory neuron-specific G-protein coupled receptors (SNSRs), are selectively expressed in subpopulations of sensory neurons involved in the perceptor of pain (Dong et al., 2001). The Mrg family in rodents consists of 50 receptors in the MrgA-MrgH subfamilies, but humans contain fewer Mrg members. MrgD (also known as TGR7) is found across species from rodents to humans, and its expression is restricted to nonpeptidergic, small-diameter, IB4+ dorsal root ganglion (DRG) neurons. MrgD binds β-alanine, but not L-alanine, which results in increased intracellular calcium and the suppression of forskolin-induced cAMP production, thus suggesting coupling to Gq and Gi proteins (Shinohara et al., 2004). β-alanine-induced activation of MrgD inhibits KCNQ2/3-mediated M-currents in cells overexpressing MrgD and KCNQ2/3 and also in cultured DRG neurons, which indicates that MrgD enhances nociceptive signaling (Crozier et al., 2007). The cloned human MrgD-expressing cell line is made in the Chem-1 host, which supports high levels of recombinant MrgD expression on the cell surface and contains optimal levels of the promiscuous G protein to couple the receptor to the calcium signaling pathway. Thus, the cell line is an ideal tool for screening for antagonists of interactions between MrgD and its ligands.

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    Contents: 2 vials of mycoplasma-free cells, 1 mL per vial.
    Storage: Vials are to be stored in liquid N2.
    Applications: Calcium Flux Assay
    Host Cell: Chem-1, an adherent cell line expressing the promiscuous G-protein, Gα15.
    Exongenous Gene Expression: Human MrgD Mas-Related cDNA (Accession Number: NM_198923- see CODING SEQUENCE below) and promiscuous G protein are expressed in a bicistronic vector
    GMO: This product contains genetically modified organisms.
    Reference 1: 1. Crozier RA et al. (2007). MrgD Activation Inhibits KCNQ/M-Currents and Contributes to Enhanced Neuronal Excitability. J. Neurosci. 27(16): 4492-4496.
    Reference 2: 2. Dong X et al. (2001) A diverse family of GPCRs expressed in specific subsets of nociceptive sensory neurons. Cell 106: 619-632.
    Reference 3: 3. Shinohara T et al. (2004) Identification of a G protein-coupled receptor specifically responsive to b-alanine. J. Biol. Chem. 279: 23559-23564.
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