MULTISCREEN™ STABLE CELL LINES

HUMAN RECOMBINANT HETERODIMER GHSR DRD2 RECEPTOR

Product Information

Catalog Number:
C1514

Lot Number:
C1514-081417

Quantity:
1 vial (2 x 106) frozen cells

Freeze Medium:
Cell Banker

Host cell:
HEK293T

Transfection:
Expression vector containing full-length human DRD2 cDNA (GenBank Accession Number NM_000795.3) with FLAG tag sequence at N-terminus and GHSR cDNA (GenBank Accession Number NM_198407.1) with MYC tag sequence at N-terminus

Recommended Storage:
Liquid nitrogen upon receiving

Propagation Medium: DMEM, 10% FBS, 1 μg/mL puromycin, 250ug/ml hygromycin

Stability:
In progress

Data Sheet

Background: The concept of dimerization of G protein-coupled receptor (GPCR) has opened a new insight regarding physiological function regulation. One of the example is heterodimerization of Ghrelin(GHSR) and dopamine(DRD2) receptor. Recent experiments have showed that GHSR:DRD2 heterodimers allosterically modifies canonical DRD2 dopamine signaling resulting in Gβγ subunit-dependent mobilization of Ca2+ independent of GHSR basal activity. Independently, agonist activation of GHSR results in coupling to Gαq and DRD2 transmits dopamine signal through Gαi/o coupling by inhibiting activity of adenylate cyclase and decreasing cAMP level in dose dependent manner.

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. Receptor expression on cell surface measured by flow cytometry (FACS) using: a. Anti- FLAG antibody, b. Anti-MYC antibody. Thin line: parental cells; thick line: receptor-expressing cells.

References:

van der Lely et al. (2004) Biological, physiological, pathophysiological, and pharmacological aspects of ghrelin. Endocr Rev 25:426-457.

Kern et al. (2013) Apo-ghrelin receptor forms heteromers with DRD2 in hypothalamic neurons and is essential for anorexigenic effects of DRD2 agonism. Neuron. 73(2): 317–332.

FOR RESEARCH USE ONLY.
All rights reserved. No part of this document may be reproduced in any form without prior permission in writing.

26219 Eden Landing Road Hayward, CA 94545 | (510) 887-0817 | info@multispaninc.com