Cloning and expression of a cDNA for the human prostacyclin receptor.
A functional cDNA for the human prostacyclin receptor was isolated from a cDNA library of CMK cells, a human megakaryocytic leukaemia cell line. The cDNA encodes a protein consisting of 386 amino acid residues with seven putative transmembrane domains and a deduced molecular weight of 40,956. [3H]Iloprost specifically bound to the membrane of CHO cells stably expressing the cDNA with a Kd of 3.3 nM. This binding was displaced by unlabelled prostanoids in the order of iloprost = cicaprost >> carbacyclin > prostaglandin E1 (PGE1) > STA2. PGE2, PGD2 and PGF 2 alpha did not inhibit it. Iloprost in a concentration-dependent manner increased the cAMP level and generated inositol trisphosphate in these cells, indicating that this human receptor can couple to multiple signal transduction pathways.
Katsuyama
M
M
Department of Physiological Chemistry, Faculty of Pharmaceutical Sciences, Kyoto University, Japan.
Sugimoto
Y
Y
Namba
T
T
Irie
A
A
Negishi
M
M
Narumiya
S
S
Ichikawa
A
A
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D001667:Binding, Competitive; D016466:CHO Cells; D003001:Cloning, Molecular; D006224:Cricetinae; D018076:DNA, Complementary / Q000737:chemistry / Q000235:genetics*; D015870:Gene Expression; D018014:Gene Transfer Techniques; D006801:Humans; D016285:Iloprost / Q000378:metabolism; D051379:Mice; D008969:Molecular Sequence Data; D044006:Receptors, Epoprostenol; D011982:Receptors, Prostaglandin / Q000737:chemistry / Q000235:genetics*; D011994:Recombinant Proteins / Q000378:metabolism; D013920:Thrombocythemia, Essential; D014407:Tumor Cells, Cultured
D018076:DNA, Complementary; D044006:Receptors, Epoprostenol; D011982:Receptors, Prostaglandin; D011994:Recombinant Proteins; D016285:Iloprost