Characterization of the human oxytocin receptor stably expressed in 293 human embryonic kidney cells

doi:10.1016/0024-3205(95)02218-8

Life Sciences

Volume 57, Issue 24, 3 November 1995, Pages 2253-2261

https://doi.org/10.1016/0024-3205(95)02218-8 Get rights and content

Abstract

The human oxytocin (OT) receptor was stably expressed in 293 embryonic kidney cells (293/OTR), characterized pharmacologically and compared to human uterine myometrial receptors. The cloned receptor is expressed at a reasonably high density (0.82 fmole/gmg protein) and exhibits high affinity for [³H]OT (K_d = 0.32 nM), similar to the value found in human myometrial tissue. The rank-order of potency for various antagonist and agonist ligands from several structural classes is also similar between the cloned and native receptor, as seen in a comparison of their inhibitory constants for [³H]OT binding. Agonist affinity at the cloned OT receptor is decreased by guanine nucleotide analogs, demonstrating functional Gprotein-coupling. The OT receptor in 293 cells, like in human myometrium, is also coupled to the inositol phosphate pathway. In 293/OTR cells, OT stimulates inositol phosphate accumulation with an EC₅₀=4.1 nM, an effect blocked by a potent and selective OT antagonist, L-366,948. Additionally, the cloned receptor in 293 cells desensitizes to high concentrations of OT, similar to the desensitization in myometrial tissue and also described for several other Gprotein-coupled receptors. These results illustrate the utility of the 293 cell line for expressing human OT receptors in an environment quite comparable to the native myometrial tissue.

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¹: Current Address: Dept. Neuropharmacology, Roche Bioscience, Palo Alto, CA

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Characterization of the human oxytocin receptor stably expressed in 293 human embryonic kidney cells

Abstract

J. Biol. Chem.

J. Biol. Chem.

J. Biol. Chem.

J. Biol. Chem.

Biochem. Biophys. Acta

Trends Pharmacol. Sci.

Am. J. Obstet. Gynecol..

Nature