Cloning and expression of the alveolar type II cell P2u-purinergic receptor.
Cloning and expression of the alveolar type II cell P2u-purinergic receptor.
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肺泡 II 型细胞 P2u 嘌呤能受体的克隆和表达。
DOI：
10.1165/ajrcmb.12.1.7811468
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发表时间：
1995
期刊：
影响因子：
--
通讯作者：
Fiedeldey,DT
中科院分区：
文献类型：
--
作者：
Rice,WR;Burton,FM;Fiedeldey,DT
三磷酸腺苷（Adenosine triphosphate， ATP）通过与肺泡II型细胞表面的p2 -嘌呤受体相互作用，调节肺泡II型细胞的表面活性剂磷脂分泌。为了进一步表征表面活性剂分泌的调节，我们克隆了II型细胞p2u -嘌呤受体，并在一个不相关的细胞系中表达了一个功能性受体。从ⅱ型细胞cDNA文库中分离得到的P2u克隆编码序列为1.1 kb，编码374个氨基酸。该蛋白与之前在神经母细胞瘤与胶质瘤杂交细胞系中发现的p2o -嘌呤受体有97%的同源性，与最近克隆的人类p2o -嘌呤受体有87%的同源性，与从鸡脑中克隆的p2o -嘌呤受体有34%的同源性。假定的II型细胞P2u蛋白包含7个跨膜结构域，具有g蛋白偶联受体的特征。II型细胞p2o -嘌呤受体核苷酸序列与NG 108-15克隆的核苷酸序列同源性为95%。然而，II型细胞cDNA在5'非翻译区也存在额外的208bp插入，这在NG 108-15克隆中不存在。利用逆转录聚合酶链反应，我们检测了两种不同大小的mRNA在不同大鼠组织中的表达。大鼠心脏、肾脏、肺、脾脏和睾丸中仅表达较大的II型细胞mRNA，脑和肝脏中未见p2o -嘌呤受体mRNA的表达。较小种类的mRNA仅在小鼠N18-TG2细胞中检测到，这些细胞也表达了较大种类的mRNA，在上述大鼠组织中发现。（摘要删节250字）
Adenosine triphosphate (ATP) regulates surfactant phospholipid secretion from alveolar type II cells by interacting with P2-purinoceptors on the alveolar type II cell surface. To further characterize regulation of surfactant secretion, we have cloned the type II cell P2u-purinoceptor and expressed a functional receptor in an unrelated cell line. The coding sequence of the P2u clone isolated from a type II cell cDNA library was 1.1 kb, encoding a putative protein of 374 amino acids. The putative protein demonstrated > 97% homology with the P2u-purinoceptor previously identified in the hybrid neuroblastoma x glioma cell line, NG 108-15, 87% homology to the recently cloned human P2u-purinoceptor, and 34% homology to the P2u-purinoceptor cloned from chicken brain. The putative type II cell P2u protein contains seven membrane-spanning domains, characteristic of G-protein-coupled receptors. The type II cell P2u-purinoceptor nucleotide sequence also demonstrated > 95% homology to the nucleotide sequence of the NG 108-15 clone. However, the type II cell cDNA also demonstrated presence of an additional 208 bp insert in the 5' untranslated region, which was not present in the NG 108-15 clone. Using reverse transcriptase polymerase chain reaction, we examined expression of the two different sizes of mRNA in various rat tissues. Only the larger type II cell mRNA was expressed in rat heart, kidney, lung, spleen, and testis, with no expression of P2u-purinoceptor mRNA noted in brain or liver. The smaller species of mRNA was only detected in mouse N18-TG2 cells, and these cells expressed a larger species as well, found in the rat tissues noted.(ABSTRACT TRUNCATED AT 250 WORDS)