ChemiScreen™ CALCIUM-OPTIMIZED STABLE CELL LINE
HUMAN RECOMBINANT M₂ MUSCARINIC ACETYLCHOLINE RECEPTOR

ChemiScreen™ CALCIUM-OPTIMIZED STABLE CELL LINE
HUMAN RECOMBINANT M₂ MUSCARINIC ACETYLCHOLINE RECEPTOR

• ChemiScreen
• Chemicon (Millipore)

2 vials

Full-length human CHRM2 cDNA encoding M₂

The muscarinic acetylcholine receptor (mAChR) family consists of five GPCRs that mediate some of the neurotransmission functions of acetylcholine in the CNS and the periphery. The M₁, M₃ and M₅ receptors couple to Gq to mobilize intracellular calcium, whereas the M₂ and M₄ receptors couple to Gi/o to inhibit cAMP production (Caulfield and Birdsall, 1998). In urinary bladder trachea and stomach, M₂ augments the function of M₃ in promoting contractility, and activation of M₂ serves to counteract relaxation induced by increased cAMP levels (Ehlert et al., 2005; Wess, 2004). In addition, the ability of mChR agonists to decrease heart rate appears to be mediated primarily by M₂. Agonists of mAChRs induce tremor, hypothermia, corticosterone release, and analgesia; each of these functions is mediated at least in part by M₂ (Wess, 2004). Chemicon's cloned human M₂-expressing cell line is made in the Chem-1 host, which supports high levels of recombinant M₂ 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 M₂ and its ligands.

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The muscarinic cholinergic receptors belong to a larger family of G protein-coupled receptors. The functional diversity of these receptors is defined by the binding of acetylcholine to these receptors and includes cellular responses such as adenylate cyclase inhibition, phosphoinositide degeneration, and potassium channel mediation. Muscarinic receptors influence many effects of acetylcholine in the central and peripheral nervous system. The muscarinic cholinergic receptor 2 is involved in mediation of bradycardia and a decrease in cardiac contractility. Multiple alternatively spliced transcript variants have been described for this gene.

FUNCTION: SwissProt: P08172 # The muscarinic acetylcholine receptor mediates various cellular responses, including inhibition of adenylate cyclase, breakdown of phosphoinositides and modulation of potassium channels through the action of G proteins. Primary transducing effect is adenylate cyclase inhibition.
SIZE: 466 amino acids; 51715 Da
SUBCELLULAR LOCATION: Cell membrane; Multi-pass membrane protein.
SIMILARITY: SwissProt: P08172 ## Belongs to the G-protein coupled receptor 1 family.

Human

EC50 for calcium mobilization by carbamoyl choline: ~ 980 nM

Chem-1

GPCR Cell Lines

Chemicon

Chem-1

Cells are frozen at 2 x 10e6 cells/mL in DMEM/20% fetal bovine serum/100 U/ml penicillin and streptomycin/10% DMSO. Cell line tests negative for mycoplasma.

M2

P08172

non-peptide

Acetylcholine (muscarinic)

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

1 mL per vial

• CHRM2
• MGC120006
• FLJ43243
• MGC120007
• HM2

GPCRs

ChemiScreen™ CALCIUM-OPTIMIZED STABLE CELL LINE
HUMAN RECOMBINANT M₂ MUSCARINIC ACETYLCHOLINE RECEPTOR

• NM_000739.2
• NM_001006626.1
• NM_001006627.1
• NM_001006628.1
• NM_001006629.1
• NM_001006630.1
• NM_001006631.1
• NM_001006632.1

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)/100U/ml each penicillin and streptomycin

2 x 10⁶ cells/mL