Expression of preprosomatostatin in heterologous cells: biosynthesis, posttranslational processing, and secretion of mature somatostatin.
Expression of preprosomatostatin in heterologous cells: biosynthesis, posttranslational processing, and secretion of mature somatostatin.
复制标题
前原生长抑素在异源细胞中的表达：成熟生长抑素的生物合成、翻译后加工和分泌。
DOI：
10.1016/0092-8674(84)90461-6
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发表时间：
1984
期刊：
影响因子：
64.5
通讯作者：
Shields,D
中科院分区：
文献类型：
--
作者：
Warren,TG;Shields,D
生长抑素是一种14个氨基酸的多肽激素，是作为更大的前体前生长抑素的一部分合成的，前生长抑素由大约120个氨基酸组成。在许多物种中，前体的整体组织结构是保守的，因为它由一个信号肽组成，紧随其后的是一个90-100个氨基酸的前区，成熟的激素位于分子的羧基末端。为了了解前肽在生成成熟激素中的作用，我们使用基因转移实验将垂钓鱼前生长抑素导入哺乳动物细胞。在这里，我们报道了一个含有前生长抑素基因克隆到VP-1晚期基因的SV40表达载体转染COS-7细胞的结果。生长抑素基因表达参数分析表明，COS细胞合成了细胞内检测到的前生长抑素；该原生长抑素被蛋白分解为成熟的生长抑素；成熟的激素由COS细胞分泌到组织培养液中。我们的结果表明，正常情况下不分泌多肽激素的COS细胞含有将前生长抑素转化为成熟激素所需的蛋白水解酶及其分泌所需的细胞装置。
Somatostatin is a 14 amino acid peptide hormone that is synthesized as part of a larger precursor, preprosomatostatin, which comprises about 120 amino acids. The overall organization of the precursor is conserved in many species in that it consists of a signal peptide followed by a proregion of 90-100 amino acids and the mature hormone is located at the carboxyl terminus of the molecule. To understand the role of the propeptide in generating the mature hormone, we have used gene-transfer experiments to introduce angler fish preprosomatostatin into mammalian cells. Here we report the results of transfection of COS-7 cells with an SV40 expression vector containing preprosomatostatin cDNA cloned into the VP-1 late gene. Analysis of the parameters of somatostatin gene expression showed that COS cells synthesized prosomatostatin, which was detected intracellularly; the prosomatostatin was proteolytically processed to mature somatostatin; and the mature hormone was secreted by the COS cells into the tissue culture medium. Our results suggest that COS cells, which do not normally secrete polypeptide hormones, contain the necessary proteolytic processing enzymes to convert preprosomatostatin to the mature hormone and the cellular apparatus necessary for its secretion.