EP2亚型1大鼠前列腺素E2受体的克隆与表达

Ken Nemoto , Carol C Pilbeam , Stephan R Bilak , Lawrence G Raisz
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引用次数: 76

摘要

前列腺素E2 (PGE2)是多种组织中有效的局部细胞生长和分化介质。根据对选择性激动剂和拮抗剂的反应,PGE2受体可分为EP1、EP2、EP3和EP4四个药理亚型。我们从一个大鼠肺cDNA文库中克隆了一个功能的大鼠EP2受体亚型cDNA。大鼠EP2受体cDNA编码357个氨基酸残基,与人和小鼠EP2受体高度同源,包含7个推测的跨膜结构域。在转染大鼠EP2 cDNA的COS-7细胞中,发现特异性的[3H]PGE2结合,解离常数为14.9 nM,未标记的PGE2和PGF2α抑制了这种结合。PGE2和一种EP2选择性激动剂butaprost可有效提高COS-7细胞转染物中的cAMP水平。Northern blot和RT-PCR分析显示EP2受体广泛分布于各组织中。通过RT-PCR和原位杂交发现EP2在胎儿长骨和颅骨中的表达高于成人,表明该受体在快速生长的骨组织中起作用。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
Molecular Cloning and Expression of a Rat Prostaglandin E2 Receptor of the EP2 Subtype 1 2

Prostaglandin E2 (PGE2) is a potent local mediator of cell growth and differentiation in various tissues. The receptors for PGE2 have been classified into four pharmacological subtypes, EP1, EP2, EP3, and EP4, based on the responses to selective agonists and antagonists. We have cloned a functional cDNA for the rat EP2 receptor subtype from a rat lung cDNA library. The rat EP2 receptor cDNA encodes 357 amino acid residues having high homology with the human and mouse EP2 receptors and containing seven putative transmembrane domains. In COS-7 cells transfected with rat EP2 cDNA, specific [3H]PGE2 binding was found with a dissociation constant of 14.9 nM, and this binding was inhibited by unlabeled PGE2 and PGF. PGE2 and butaprost, an EP2 selective agonist, were effective in increasing the cAMP level in the COS-7 cell transfectants. Northern blot and RT-PCR analysis showed widespread distribution of the EP2 receptor in various tissues. Higher EP2 expression was found in fetal long bones and calvariae than in adult by RT-PCR and in situ hybridization, suggesting a role for this receptor in rapidly growing skeletal tissue.

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