Cloning of a novel human prostaglandin receptor with characteristics of the pharmacologically defined EP2 subtype

J. W. Regan, T. J. Bailey, D. J. Pepperl, K. L. Pierce, A. M. Bogardus, J. E. Donello, C. E. Fairbairn, K. M. Kedzie, D. F. Woodward, D. W. Gil

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325 Scopus citations

Abstract

A cDNA that when expressed has the binding and functional characteristics of the pharmacologically defined EP2 prostaglandin (PG) receptor [Cardiovasc. Drug Rev. 11:165-179 (1993)] has been cloned from a human placenta library. This clone, known as Hup-4, encodes a protein of 358 amino acids that has only ~30% overall identity with other PG receptors, including mouse and human clones that have been designated as EP2 receptors [J. Biol. Chem. 268:7759-7762 (1993); Biochem. Biophys. Res. Commun. 197:263-270 (1993)]. In COS-7 cells transfected with Hup-4, PGE2 stimulated the formation of cAMP with an EC50 of ~50 nM. The EP2-selective agonists AH13205 and butaprost were also active, with EC50 values in the range of 2- 6 μM. The order of potency of PGs for competition with binding of [3H]PGE2 to membranes prepared from COS-7 cells transfected with Hup-4 was PGE2 ≥ PGE1 > 16,16-dimethyl-PGE2 ≥ 11-deoxy-PGE1 > butaprost > AH13205 > 19(R)- OH-PGE2. Natural PGs and analogues that are selective for the FP (PGF(2α)), DP (PGD2), EP1 (sulprostone), EP3 (MB 28767), and EP4 (1-OH-PGE1) receptors were inactive or competed poorly with the binding of [3H]PGE2 (<50% displacement of specific binding at 10 μM). Northern blot analysis showed the presence of a Hup-4 message of ~3.1 kilobases in mRNA from human lung and placenta. Reverse transcription-polymerase chain reaction studies also indicated that Hup-4 is probably expressed in human uterus and in HL-60 (human promyelocytic leukemia) cells. Our findings suggest that Hup-4 encodes the pharmacologically defined EP2 receptor, whereas the mouse and human cDNAs previously classified as EP2 may represent another EP receptor subtype or the recently defined EP4 subtype [Prostaglandins 47:151-168 (1994)].

Original languageEnglish (US)
Pages (from-to)213-220
Number of pages8
JournalMolecular pharmacology
Volume46
Issue number2
StatePublished - Aug 1 1994

ASJC Scopus subject areas

  • Molecular Medicine
  • Pharmacology

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    Regan, J. W., Bailey, T. J., Pepperl, D. J., Pierce, K. L., Bogardus, A. M., Donello, J. E., Fairbairn, C. E., Kedzie, K. M., Woodward, D. F., & Gil, D. W. (1994). Cloning of a novel human prostaglandin receptor with characteristics of the pharmacologically defined EP2 subtype. Molecular pharmacology, 46(2), 213-220.