大鼠肾素在哺乳动物细胞中的表达及其纯化。

Expression of rat renin in mammalian cells and its purification.

作者信息

Yamauchi T, Suzuki F, Takahashi A, Tsutsumi I, Hori H, Watanabe T, Ishizuka Y, Nakamura Y, Murakami K

机构信息

Institute of Applied Biochemistry, University of Tsukuba, Ibaraki, Japan.

出版信息

Clin Exp Hypertens A. 1992;14(3):377-92. doi: 10.3109/10641969209036196.

DOI:10.3109/10641969209036196

PMID:1600638

Abstract

Rat renin cDNA was transfected into COS-7 and Chinese hamster ovary (CHO) cells and expressed under the control of the Simian Virus 40 early promoter. Conditioned media of the transfected cells showed renin activity only after trypsin treatment, suggesting prorenin was secreted into the medium. From the trypsinized serum-free culture of the transfected CHO cells active renin was purified to homogeneity by a simple three-step procedure. The active renin had similar specific activity, molecular weight, Km, pH optimum, and isoelectric point compared to native renin. The amino-terminal sequence was the same as that deduced from the renin cDNA. This suggests that the recombinant rat renin is similar to kidney renin in many respects, and is easily obtained by the present procedures.

摘要

大鼠肾素cDNA被转染到COS - 7细胞和中国仓鼠卵巢（CHO）细胞中，并在猿猴病毒40早期启动子的控制下表达。转染细胞的条件培养基仅在胰蛋白酶处理后显示出肾素活性，这表明前肾素被分泌到培养基中。通过一个简单的三步程序，从经胰蛋白酶处理的转染CHO细胞无血清培养物中纯化出活性肾素，使其达到同质。与天然肾素相比，活性肾素具有相似的比活性、分子量、米氏常数、最适pH值和等电点。其氨基末端序列与从肾素cDNA推导出来的序列相同。这表明重组大鼠肾素在许多方面与肾脏肾素相似，并且通过目前的程序很容易获得。