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

John 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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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 1994

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Prostaglandin Receptors
Organism Cloning
Dinoprostone
Alprostadil
COS Cells
Placenta
16,16-Dimethylprostaglandin E2
Complementary DNA
Clone Cells
Synthetic Prostaglandins
Prostaglandin D2
Prostaglandins F
Northern Blotting
Reverse Transcription
Uterus
Prostaglandins
Leukemia
Amino Acids
Polymerase Chain Reaction
Lung

ASJC Scopus subject areas

  • Pharmacology

Cite this

Regan, J. W., Bailey, T. J., Pepperl, D. J., Pierce, K. L., Bogardus, A. M., Donello, J. E., ... 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.

Cloning of a novel human prostaglandin receptor with characteristics of the pharmacologically defined EP2 subtype. / Regan, John 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.

In: Molecular Pharmacology, Vol. 46, No. 2, 08.1994, p. 213-220.

Research output: Contribution to journalArticle

Regan, JW, Bailey, TJ, Pepperl, DJ, Pierce, KL, Bogardus, AM, Donello, JE, Fairbairn, CE, Kedzie, KM, Woodward, DF & Gil, DW 1994, 'Cloning of a novel human prostaglandin receptor with characteristics of the pharmacologically defined EP2 subtype', Molecular Pharmacology, vol. 46, no. 2, pp. 213-220.
Regan, John 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. / Cloning of a novel human prostaglandin receptor with characteristics of the pharmacologically defined EP2 subtype. In: Molecular Pharmacology. 1994 ; Vol. 46, No. 2. pp. 213-220.
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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)].",
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