Cloning and expression of a cDNA for mouse prostaglandin E receptor EP2 subtype.
A functional cDNA clone encoding mouse EP2 subtype of prostaglandin (PG) E receptor was isolated from a mouse cDNA library by cross-hybridization with the mouse EP3 subtype PGE receptor cDNA. The mouse EP2 receptor consists of 513 amino acid residues with putative seven-transmembrane domains. In contrast to EP3 receptor, this receptor possesses long third intracellular loop and carboxyl-terminal tail. [3H] PGE2 specifically bound to the membrane of mammalian COS cells transfected with the cDNA. The binding to the membrane was displaced with unlabeled PG in the order of PGE2 = PGE1 >> iloprost > or = PGF2 alpha > or = PGD2. The binding was also inhibited by misoprostol, an EP2 and EP3 agonist, but not by sulprostone, an EP1 and EP3 agonist, and SC-19220, an EP1 antagonist. PGE2 markedly increased cAMP level in COS cells transfected with the cDNA. These results suggest that this receptor is EP2 subtype. Northern blot analysis demonstrated that the EP2 mRNA is widely expressed in various tissues, the abundant expression being observed in ileum, thymus, and mastocytoma P-815 cells.
Department of Physiological Chemistry, Faculty of Pharmaceutical Sciences, Kyoto University, Japan.
A
A
Honda
Y
Y
Sugimoto
T
T
Namba
A
A
Watabe
A
A
Irie
M
M
Negishi
S
S
Narumiya
A
A
Ichikawa
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D001667:Binding, Competitive; D015152:Blotting, Northern; D002460:Cell Line; D003001:Cloning, Molecular; D000242:Cyclic AMP / Q000378:metabolism; D004247:DNA / Q000235:genetics* / Q000302:isolation & purification; D015342:DNA Probes; D015232:Dinoprostone / Q000378:metabolism* / Q000494:pharmacology; D015723:Gene Library; D007700:Kinetics; D012515:Mast-Cell Sarcoma; D051379:Mice; D008969:Molecular Sequence Data; D009928:Organ Specificity; D016133:Polymerase Chain Reaction / Q000379:methods; D012333:RNA, Messenger / Q000302:isolation & purification / Q000378:metabolism; D011982:Receptors, Prostaglandin / Q000235:genetics* / Q000378:metabolism*; D018078:Receptors, Prostaglandin E; D017482:Receptors, Thromboxane / Q000235:genetics; D017386:Sequence Homology, Amino Acid; D014162:Transfection; D014407:Tumor Cells, Cultured
D015342:DNA Probes; D012333:RNA, Messenger; D011982:Receptors, Prostaglandin; D018078:Receptors, Prostaglandin E; D017482:Receptors, Thromboxane; D004247:DNA; D000242:Cyclic AMP; D015232:Dinoprostone