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Molecular cloning and expression of a prostaglandin E 2 receptor of the EP 3β subtype from rat hepatocytes
Author(s) -
Neuschäfer-Rube Frank,
DeVries Christa,
Hänecke Kristina,
Jungermann Kurt,
Püschel Gerhard P.
Publication year - 1994
Publication title -
febs letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.593
H-Index - 257
eISSN - 1873-3468
pISSN - 0014-5793
DOI - 10.1016/0014-5793(94)00837-x
Subject(s) - receptor , microbiology and biotechnology , complementary dna , biology , transfection , forskolin , prostaglandin , agonist , endocrinology , biochemistry , gene
Rat hepatocytes have previously been reported to possess prostaglandin E 2 receptors of the EP 3 ‐type (EP 3 ‐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 PGE 2 with an apparent K d of 15 nM. PGE 2 > PGF 2α > PGD 2 competed for [ 3 H]PGE 2 binding sites as did the EP 3 receptor agonists M&B 28767 = sulprostone > misoprostol but not the EP 1 receptor antagonist SC 19220. In stably transfected CHO cells M&B 28767 > sulprostone = PGE 2 > 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.