MOLECULAR CHARACTERIZATION OF A MOUSE PROSTAGLANDIN-D RECEPTOR AND FUNCTIONAL EXPRESSION OF THE CLONED GENE

被引:207
作者
HIRATA, M [1 ]
KAKIZUKA, A [1 ]
AIZAWA, M [1 ]
USHIKUBI, F [1 ]
NARUMIYA, S [1 ]
机构
[1] KYOTO UNIV,FAC MED,DEPT PHARMACOL,KYOTO 606,JAPAN
关键词
PROSTAGLANDIN D-2; G PROTEIN-COUPLED RECEPTOR; GENE CLONING; MESSENGER-RNA DISTRIBUTION;
D O I
10.1073/pnas.91.23.11192
中图分类号
O [数理科学和化学]; P [天文学、地球科学]; Q [生物科学]; N [自然科学总论];
学科分类号
07 ; 0710 ; 09 ;
摘要
Prostanoid receptors belong to the family of G protein-coupled receptors with seven transmembrane domains. By taking advantage of nucleotide sequence homology among the prostanoid receptors, we have isolated and identified a cDNA fragment and its gene encoding a mouse prostaglandin (PG) D receptor by reverse transcription polymerase chain reaction and gene cloning. This gene codes for a polypeptide of 357 amino acids, with a calculated molecular weight of 40,012. The deduced amino acid sequence has a high degree of similarity with the mouse PGI receptor and the EP(2) subtype of the PGE receptor, which together form a subgroup of the prostanoid receptors. Chinese hamster ovary cells stably expressing the gene showed a single class of binding sites for [H-3]PGD(2) with a K-d of 40 nM. This binding was displaced by unlabeled ligands in the following order: PGD(2) > BW 245C (a PGD agonist) > BW A868C (a PGD antagonist) > STA(2) (a thromboxane A(2) agonist). PGE(2), PGF(2 alpha), and iloprost showed little displacement activity at concentrations up to 10 mu M. PGD(2) and BW 245C also increased cAMP levels in Chinese hamster ovary cells expressing the receptor, in a concentration-dependent manner. BW A868C showed a partial agonist ac tivity in the cAMP assay. Northern blotting analysis with mouse poly (A)(+) RNA identified a major mRNA species of 3.5 kb that was most abundantly expressed in the ileum, followed by lung, stomach, and uterus.
引用
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页码:11192 / 11196
页数:5
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