Cloning and expression of a cDNA for mouse prostaglandin E receptor EP3 subtype.
A functional cDNA clone for mouse EP3 subtype of prostaglandin (PG) E receptor was isolated from a mouse cDNA library using polymerase chain reaction based on the sequence of the human thromboxane A2 receptor and cross-hybridization screening. The mouse EP3 receptor consists of 365 amino acid residues with putative seven-transmembrane domains. The sequence revealed significant homology to the human thromboxane A2 receptor. Ligand binding studies using membranes of COS cells transfected with the cDNA revealed specific [3H]PGE2 binding. The binding was displaced with unlabeled PGs in the order of PGE2 = PGE1 greater than iloprost greater than PGF2 alpha greater than PGD2. The EP3-selective agonists, M&B 28,767 or GR 63799X, potently competed for the [3H]PGE2 binding, but no competition was found with EP1- or EP2-selective ligands. PGE2 and M&B 28,767 decreased forskolin-induced cAMP formation in a concentration-dependent manner in Chinese hamster ovary cells permanently expressing the cDNA. Northern blot analysis demonstrated that the EP3 mRNA is expressed abundantly in kidney, uterus, and mastocytoma P-815 cells and in a lesser amount in brain, thymus, lung, heart, stomach, and spleen.
Department of Physiological Chemistry, Faculty of Pharmaceutical Sciences, Kyoto University, Japan.
Y
Y
Sugimoto
T
T
Namba
A
A
Honda
Y
Y
Hayashi
M
M
Negishi
A
A
Ichikawa
S
S
Narumiya
D000527:Alprostadil / Q000031:analogs & derivatives / Q000494:pharmacology; D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D015152:Blotting, Northern; D016466:CHO Cells; D003001:Cloning, Molecular; D005576:Colforsin / Q000494:pharmacology; D006224:Cricetinae; D000242:Cyclic AMP / Q000378:metabolism; D004247:DNA / Q000235:genetics*; D015232:Dinoprostone / Q000378:metabolism*; D015870:Gene Expression; D006801:Humans; D051379:Mice; D008969:Molecular Sequence Data; D011453:Prostaglandins / Q000378:metabolism; D011459:Prostaglandins E, Synthetic / Q000494:pharmacology; D012313:RNA / Q000235:genetics; D011982:Receptors, Prostaglandin / Q000187:drug effects / Q000235:genetics*; D017482:Receptors, Thromboxane; D012689:Sequence Homology, Nucleic Acid; D013931:Thromboxanes / Q000378:metabolism; D014162:Transfection
D011453:Prostaglandins; D011459:Prostaglandins E, Synthetic; D011982:Receptors, Prostaglandin; D017482:Receptors, Thromboxane; D013931:Thromboxanes; C067522:GR 63799X; D005576:Colforsin; D012313:RNA; C032405:11-deoxy-16-phenoxy-17,18,19,20-tetranorprostaglandin E1; D004247:DNA; D000242:Cyclic AMP; D000527:Alprostadil; D015232:Dinoprostone