Cloning and expression of three isoforms of the human EP3 prostanoid receptor.
Functional cDNA clones coding for three isoforms of the human prostaglandin E receptor EP3 subtype have been isolated from kidney and uterus cDNA libraries. The three isoforms, designated hEP3-I, hEP3-II and hEP3-III, have open reading frames corresponding to 390, 388 and 365 amino acids, respectively. They differ only in the length and amino acid composition of their carboxy-terminal regions, beginning at position 360. The human EP3 receptor has seven predicted transmembrane spanning domains and therefore belongs to the G-protein-coupled receptor family. The rank order of potency for prostaglandins and related analogs in competition for [3H]PGE2 specific binding to membranes prepared from transfected COS cells was comparable for all three isoforms, and as predicted for the EP3 receptor, with PGE2 = PGE1 >> PGF2 alpha = iloprost > PGD2 >> U46619. In addition, the EP3-selective agonist MB28767 was a potent competing ligand with an IC50 value of 0.3 nM, whereas the EP1-selective antagonist AH6909 gave IC50 values of 2-7 microM and the EP2-selective agonist butaprost was inactive. In summary, we have cloned three isoforms of the human EP3 receptor having comparable ligand binding properties.
Adam
M
M
Department of Biochemistry and Molecular Biology, Merck Frosst Centre for Therapeutic Research, Quebec, Canada.
Boie
Y
Y
Rushmore
T H
TH
Müller
G
G
Bastien
L
L
McKee
K T
KT
Metters
K M
KM
Abramovitz
M
M
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D002417:Cattle; D002460:Cell Line; D003001:Cloning, Molecular; D015342:DNA Probes; D015870:Gene Expression; D006801:Humans; D051379:Mice; D008969:Molecular Sequence Data; D008970:Molecular Weight; D016133:Polymerase Chain Reaction; D051381:Rats; D018078:Receptors, Prostaglandin E / Q000737:chemistry / Q000235:genetics*; D017386:Sequence Homology, Amino Acid; D014162:Transfection
D015342:DNA Probes; D018078:Receptors, Prostaglandin E