Molecular cloning and expression of a prostaglandin E2 receptor of the EP3β subtype from rat hepatocytes

FEBS Letters

Hänecke, Kristina¹  Jungermann, Kurt¹  Neuschäfer-Rube, Frank¹  Püschel, Gerhard P.¹  DeVries, Christa¹ 

[1] Institut für Biochemie und Molekulare Zellbiologie, Humboldtallee 23, Georg-August-Universität, Göttingen, Germany

关键词: Prostaglandin receptor;    Hepatocyte (rat);    Molecular cloning and expression;    CHO cells;    chinese hamster ovary cells;    DMEM;    Dulbeeco's modified eagle medium;    FCS;    fetal calf serum;    HEK cells;    human embryonal kidney cells;    IBMX;    3-isobutyl-1-methylxanthine;    PCR;    polymerase chain reaction;    PG;    prostaglandin;   

DOI  :  10.1016/0014-5793(94)00837-X

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

Rat hepatocytes have previously been reported to possess prostaglandin E₂ receptors of the EP₃-type (EP₃-receptors) that inhibit glucagon-stimulated glycogenolysis by decreasing cAMP. Here, the isolation of a functional EP_3β receptor cDNA clone from a rat hepatocyte cDNA library is reported. This clone can be translated into a 362-amino-acid protein, that displays over 95% homology to the EP_3β receptor from mouse mastocytoma. The amino- and carboxy-terminal region of the protein are least conserved. Transiently transfected HEK 293 cells expressed a single binding site for PGE₂ with an apparent K _d of 15 nM. PGE₂ > PGF_2α > PGD₂ competed for [³H]PGE₂ binding sites as did the EP₃ receptor agonists M&B 28767 = sulprostone > misoprostol but not the EP₁ receptor antagonist SC 19220. In stably transfected CHO cells M&B 28767 > sulprostone = PGE₂ > misoprostol > PGF_2α inhibited the forskolin-elicited cAMP formation. Thus, the characteristics of the EP_3β receptor of rat hepatocytes closely resemble those of the EP_3β receptor of mouse mastocytoma.

【 授权许可】

Unknown