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一种禽源G蛋白偶联P2Y受体的分子克隆与表达

Molecular cloning and expression of an avian G protein-coupled P2Y receptor.

作者信息

Boyer J L, Waldo G L, Harden T K

机构信息

Department of Pharmacology, University of North Carolina School of Medicine, Chapel Hill 27599, USA.

出版信息

Mol Pharmacol. 1997 Dec;52(6):928-34. doi: 10.1124/mol.52.6.928.

Abstract

A family of G protein-coupled P2Y receptors that are activated by adenine and uridine nucleotides has been identified recently. Degenerate primers based on conserved sequences in these P2Y receptors were used to amplify turkey DNA, which was used to isolate the complete coding sequence of a cDNA that encodes a novel G protein-coupled receptor. Stable expression of this avian cDNA in 1321N1 human astrocytoma cells resulted in the conveyance of marked inositol phosphate responses to various nucleotides. Although this cloned avian receptor exhibited its highest homology to the previously cloned mammalian P2Y4 receptor, its pharmacological selectivity was not consistent with the avian receptor's being a species homologue of the P2Y4 receptor. That is, whereas the P2Y4 receptor is selectively activated by UTP and is not activated by ATP or Ap4A, the novel avian receptor was potently activated by ATP and Ap4A as well as by UTP. Taken together, these results describe the identification of an avian phospholipase C-coupled P2Y receptor that, like the mammalian P2Y2 receptor, is activated by both adenine and uridine nucleotides.

摘要

最近已鉴定出一类由腺嘌呤和尿苷核苷酸激活的G蛋白偶联P2Y受体家族。基于这些P2Y受体保守序列的简并引物用于扩增火鸡DNA,该DNA用于分离编码新型G蛋白偶联受体的cDNA的完整编码序列。该禽类cDNA在1321N1人星形细胞瘤细胞中的稳定表达导致对各种核苷酸产生明显的肌醇磷酸反应。尽管这种克隆的禽类受体与先前克隆的哺乳动物P2Y4受体具有最高的同源性,但其药理学选择性与该禽类受体作为P2Y4受体的物种同源物不一致。也就是说,P2Y4受体被UTP选择性激活,不被ATP或Ap4A激活,而新型禽类受体被ATP、Ap4A以及UTP有效激活。综上所述,这些结果描述了一种禽类磷脂酶C偶联P2Y受体的鉴定,该受体与哺乳动物P2Y2受体一样,可被腺嘌呤和尿苷核苷酸激活。

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