ChemiScreen™ Calcium-Optimized Stable Cell Line Human Recombinant SST3 Somatostatin Receptor

ChemiScreen™ Calcium-Optimized Stable Cell Line Human Recombinant SST3 Somatostatin Receptor

• ChemiScreen
• Chemicon (Millipore)

2 vials

Somatostatin is a 14 or 28 amino acid regulatory peptide that inhibits hormone secretion from the pituitary, pancreas, and other endocrine sites. A family of 6 GPCRs, SST₁, SST_2A, SST_2B, SST₃, SST₄ and SST₅, mediate the biological activity of somatostatins. The somatostatin receptors couple to G_i to inhibit cAMP production, and also increase MAP kinase signalling. Several tumors have been shown to overexpress somatostatin receptors, and binding of somatostatin to these tumor cells stimulates or inhibits proliferation, depending on the receptor subtypes expressed (Olias et al., 2004). However, SST₃ appears to promote apoptosis, and expression of SST₃ was found to be lower in gastric cancer cells than in normal gastric mucosa, in proportion to susceptibility to apoptosis induced by somatostatin analogs (Sharma et al., 1996; Hu et al., 2004). In addition, SST₃ is expressed in smooth muscle cells and mediates gastrointestinal contractions. Chemicon's cloned human SST₃-expressing cell line is made in the Chem-1 host, which supports high levels of recombinant SST₃ 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 SST₃ and its ligands.

Calcium flux assay, ligand binding assays

Human

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Somatostatin acts at many sites to inhibit the release of many hormones and other secretory proteins. The biological effects of somatostatin are probably mediated by a family of G protein-coupled receptors that are expressed in a tissue-specific manner. SSTR3 is a member of the superfamily of receptors having seven transmembrane segments and is expressed in highest levels in brain and pancreatic islets. SSTR3 is functionally coupled to adenylyl cyclase.

FUNCTION: SwissProt: P32745 # Receptor for somatostatins-14 and -28. This receptor is coupled via pertussis toxin sensitive G proteins to inhibition of adenylyl cyclase.
SIZE: 418 amino acids; 45847 Da
SUBCELLULAR LOCATION: Cell membrane; Multi-pass membrane protein (By similarity). Note=Internalized into endoplasmic vesicles upon somatostatin-stimulation (By similarity).
TISSUE SPECIFICITY: Brain, pituitary and pancreas.
PTM: Phosphorylated. Phosphorylation increases upon somatostatin binding (By similarity).
SIMILARITY: SwissProt: P32745 ## Belongs to the G-protein coupled receptor 1 family.

Human

SPECIFICATIONS: EC50 for calcium mobilization by Somatostatin-14: ~ 3.9nM HOST CELLS: Chem-1, an adherent cell line expressing the promiscuous G-protein, Gα15. TRANSFECTION: Full-length human SSTR3 cDNA encoding SST3 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

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.

sst3

P32745

peptide

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 37°C 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.

Somatostatin

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A

2x10⁶ cells/vial

• SSTR3
• SS3R
• OTTHUMP00000028737
• SSR-28

GPCRs

ChemiScreen™ Calcium-Optimized Stable Cell Line Human Recombinant SST3 Somatostatin Receptor

NM_001051.2