THE MOUSE PROSTAGLANDIN-E RECEPTOR EP(2) SUBTYPE - CLONING, EXPRESSION, AND NORTHERN BLOT ANALYSIS
THE MOUSE PROSTAGLANDIN-E RECEPTOR EP(2) SUBTYPE - CLONING, EXPRESSION, AND NORTHERN BLOT ANALYSIS
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DOI：
10.1016/0014-5793(95)00966-d
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发表时间：
1995-09-25
期刊：
影响因子：
3.5
通讯作者：
ICHIKAWA, A
中科院分区：
文献类型：
--
作者：
KATSUYAMA, M;NISHIGAKI, N;ICHIKAWA, A
从小鼠 cDNA 文库中分离出小鼠前列腺素 (PG) E 受体 EP(2) 亚型的功能 cDNA 克隆。小鼠 EP(2) 受体由 362 个氨基酸残基和 7 个假定的跨膜结构域组成。 [H-3]PGE(2) 与稳定表达克隆受体的中国仓鼠卵巢细胞膜特异性结合。这种结合被未标记的前列腺素取代，其顺序为 PGE(2) = PGE(1)，远大于伊洛前列素（一种稳定的 PGI(2) 激动剂） > PGF(2α) > PGD(2)。结合mas也被布他前列素（一种EP（2）激动剂）抑制，并在较小程度上被M&B 28767（一种EP（3）激动剂）抑制，但不被磺前列酮（一种EP（1）和EP（3）激动剂）或SC-19220（一种EP（1）拮抗剂）抑制，PGE（2）和布他前列素增加了中国仓鼠的cAMP水平卵巢细胞以浓度依赖的方式。 Northern印迹分析显示EP(2) mRNA在子宫中表达最丰富，其次是脾、肺、胸腺、回肠、肝和胃。
A functional cDNA clone for the mouse prostaglandin (PG) E receptor EP(2) subtype was isolated from a mouse cDNA library, The mouse EP(2) receptor consists of 362 amino acid residues with seven putative transmembrane domains. [H-3]PGE(2) bound specifically to the membrane of Chinese hamster ovary cells stably expressing the cloned receptor. This binding was displaced by unlabeled prostanoids in the order of PGE(2) = PGE(1) much greater than iloprost, a stable PGI(2) agonist > PGF(2 alpha) > PGD(2). Binding mas also inhibited by butaprost (an EP(2) agonist) and to a lesser extent by M&B 28767 (an EP(3) agonist), but not by sulprostone (an EP(1) and EP(3) agonist) or SC-19220 (an EP(1) antagonist), PGE(2) and butaprost increased the cAMP level in the Chinese hamster ovary cells in a concentration-dependent manner. Northern blot analysis revealed that EP(2) mRNA is expressed most abundantly in the uterus, followed by the spleen, lung, thymus, ileum, liver, and stomach.