ChemiScreen™ Human recombinant Motilin Receptor, Calcium-Optimized Stable Cell Line

ChemiScreen™ Human recombinant Motilin Receptor, Calcium-Optimized Stable Cell Line

• ChemiScreen
• Chemicon (Millipore)

2 vials

TRANSFECTION: Full-length human MLNR cDNA encoding the Motilin Receptor.

Motilin is a 22 amino acid peptide that potently stimulates gastrointestinal contractility. The biological effects of motilin are mediated by a G_q-coupled seven transmembrane protein, currently termed motilin receptor (MR), that shares significant sequence similarity with the ghrelin receptor (Feighner et al., 1999). The motilin receptor is also activated by the antibiotic erythromycin; this interaction appears to mediate some of the gastrointestinal side effects of erythromycin. Although motilides (non-antibiotic derivatives of erythromycin) such as ABT-229 have been investigated for treatment of diabetic gastroporesis, the effectiveness has been limited by tachyphylaxis (decreased response to ligand) resulting from receptor downregulation (Thielemans et al., 2005). Agonists of the motilin receptor with reduced densensitization activity remain a potential treatment for disorders of gastric motility. Chemicon's cloned human Motilin Receptor-expressing cell line is made in the Chem-1 host, which supports high levels of recombinant Motilin Receptor expression on the cell surface and contains high levels of the promiscuous G protein Gα15 to couple the receptor to the calcium signaling pathway. Thus, the cell line is an ideal tool for screening for antagonists of interactions between the Motilin Recptor and its ligands.

Calcium flux assay, ligand binding assays

Human

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FUNCTION: SwissProt: O43193 # Receptor for motilin.
SIZE: 412 amino acids; 45344 Da
SUBCELLULAR LOCATION: Cell membrane; Multi-pass membrane protein.
TISSUE SPECIFICITY: Expressed only in thyroid, stomach, and bone marrow.
SIMILARITY: SwissProt: O43193 ## Belongs to the G-protein coupled receptor 1 family.

Human

SPECIFICATIONS: EC50 for calcium mobilization by Motilin: ~ 1.8 nM

Chem-1

GPCR Cell Lines

Chemicon

Chem-1

Cells are frozen at 2 x 10⁶ cells/mL in DMEM/20% fetal bovine serum/100 U/ml penicillin and streptomycin/10% DMSO. Cell line tests negative for mycoplasma.

motilin receptor / MTLR / GPR38

Cell Line

O43193

peptide

Motilin

Place cells in liquid nitrogen immediately upon receipt. Maintain frozen in liquid nitrogen for up to 5 years. Thaw cells rapidly by removing from liquid nitrogen and immediately immersing in a 37ºC water bath. Immediately after ice has thawed, sterilize the exterior of the vial with 70% ethanol. Transfer contents of the vial to a T75 flask containing 20 mL growth media, and place in a humidified 37°C incubator with 5% CO2.. After 8-24 h, cells will adhere to the plate, at which time the media should be replaced to remove residual DMSO. Cells are passaged by washing with Ca⁺⁺ and Mg⁺⁺-free HBSS (10 mL/T75), incubating with 0.05% trypsin/0.2 g/L EDTA (1 mL/T75) for 5-10 minutes at 37°C, and rapping the side of the flask to dislodge the cells. Neutralize the trypsin by addition of 4 volumes growth media. Cells are typically passaged 1:10 with every 3-4 days, and should be passaged at least once after thawing prior to use in calcium flux assays.

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A

• MLNR
• GPR38
• MTLR
• MTLR1

GPCRs

ChemiScreen™ Human recombinant Motilin Receptor, Calcium-Optimized Stable Cell Line

NM_001507.1

GROWTH MEDIA: DMEM containing 4.5 g/L glucose/10% heat inactivated fetal bovine serum/1x nonessential amino acids/10 mM HEPES/0.25 mg/ml Geneticin (G418)/100 U/ml each penicillin and streptomycin.

2 x 10⁶ cells/mL