Cloning of a novel human prostaglandin receptor with characteristics of the pharmacologically defined EP2 subtype.
Cloning of a novel human prostaglandin receptor with characteristics of the pharmacologically defined EP2 subtype.
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发表时间：
1994-08
影响因子：
3.6
通讯作者：
J. Regan;T. Bailey;D. J. Pepperl;K. L. Pierce;A. Bogardus;J. Donello;C. E. Fairbairn;K. Kedzie;D. Woodward;D. Gil
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作者：
J. Regan;T. Bailey;D. J. Pepperl;K. L. Pierce;A. Bogardus;J. Donello;C. E. Fairbairn;K. Kedzie;D. Woodward;D. Gil
一种cDNA，其在表达时具有Ep定义的EP 2前列腺素（PG）受体的结合和功能特征[Ep vasc. Drug Rev. 11：165-179（1993）]已从人胎盘文库中克隆。该克隆称为Hup-4，编码358个氨基酸的蛋白质，其与其它PG受体仅具有约30%的总体同一性，所述其它PG受体包括已被指定为EP 2受体的小鼠和人克隆[J. Biol. Chem. 268：7759-7762（1993）; Biochem. Biophys. Res. Commun. 197：263-270（1993）]。在转染Hup-4的COS-7细胞中，PGE 2刺激cAMP的形成，EC 50约为50 nM。EP 2选择性激动剂AH 13205和布他前列素也有活性，EC 50值在2-6 μ M范围内。PGs竞争[3 H] PGE 2与Hup-4转染COS-7细胞膜结合的效力顺序为PGE 2>或= PGE 1> 16，16-二甲基-PGE 2>或= 11-脱氧-PGE 1>布他前列素> AH 13205> 19（R）-OH-PGE 2。对FP（PGF 2a）、DP（PGD 2）、EP 1（硫前列酮）、EP 3（MB 28767）和EP 4（1-OH-PGE 1）受体具有选择性的天然PG和类似物无活性或与[3 H] PGE 2的结合竞争性差（在10 μ M下特异性结合的置换< 50%）。北方印迹分析显示，在人肺和胎盘的mRNA中存在约3.1个Hup-4信使。逆转录-聚合酶链反应研究还表明，Hup-4可能在人子宫和HL-60（人早幼粒细胞白血病）细胞中表达。我们的发现表明，Hup-4编码EEP 2受体，而先前分类为EP 2的小鼠和人cDNA可能代表另一种EP受体亚型或最近定义的EP 4亚型[Prostaglandins 47：151-168（1994）]。
A cDNA that when expressed has the binding and functional characteristics of the pharmacologically defined EP2 prostaglandin (PG) receptor [Cardiovasc. Drug Rev. 11:165-179 (1993)] has been cloned from a human placenta library. This clone, known as Hup-4, encodes a protein of 358 amino acids that has only approximately 30% overall identity with other PG receptors, including mouse and human clones that have been designated as EP2 receptors [J. Biol. Chem. 268:7759-7762 (1993); Biochem. Biophys. Res. Commun. 197:263-270 (1993)]. In COS-7 cells transfected with Hup-4, PGE2 stimulated the formation of cAMP with an EC50 of approximately 50 nM. The EP2-selective agonists AH13205 and butaprost were also active, with EC50 values in the range of 2-6 microM. The order of potency of PGs for competition with binding of [3H]PGE2 to membranes prepared from COS-7 cells transfected with Hup-4 was PGE2 > or = PGE1 > 16,16-dimethyl-PGE2 > or = 11-deoxy-PGE1 > butaprost > AH13205 > 19(R)-OH-PGE2. Natural PGs and analogues that are selective for the FP (PGF2a), DP (PGD2), EP1 (sulprostone), EP3 (MB 28767), and EP4 (1-OH-PGE1) receptors were inactive or competed poorly with the binding of [3H]PGE2 (< 50% displacement of specific binding at 10 microM). Northern blot analysis showed the presence of a Hup-4 message of approximately 3.1 kilobases in mRNA from human lung and placenta. Reverse transcription-polymerase chain reaction studies also indicated that Hup-4 is probably expressed in human uterus and in HL-60 (human promyelocytic leukemia) cells. Our findings suggest that Hup-4 encodes the pharmacologically defined EP2 receptor, whereas the mouse and human cDNAs previously classified as EP2 may represent another EP receptor subtype or the recently defined EP4 subtype [Prostaglandins 47:151-168 (1994)].