昆虫细胞中单链尿激酶型纤溶酶原激活剂的分泌。

Secretion of single-chain urokinase-type plasminogen activator from insect cells.

作者信息

King L A, Kaur K, Mann S G, Lawrie A M, Steven J, Ogden J E

机构信息

School of Biological and Molecular Sciences, Oxford Polytechnic, Headington, U.K.

出版信息

Gene. 1991 Oct 15;106(2):151-7. doi: 10.1016/0378-1119(91)90194-g.

DOI:10.1016/0378-1119(91)90194-g

PMID:1834526

Abstract

A cDNA encoding human urokinase-type plasminogen activator was inserted downstream from the polyhedrin promoter of the baculovirus Autographa californica nuclear polyhedrosis virus. A protein of similar Mr to urokinase (UK) was synthesized and approx. 90% was secreted from recombinant virus-infected Spodoptera frugiperda cells. Zymography and Western blotting analysis of the insect-derived protein demonstrated that it was comprised solely of the high-Mr form of UK. No low-Mr UK was detected. Amidolytic activity assays showed that 96% of the insect cell-derived UK was in the single-chain proenzyme form. The yield of UK from insect cells was 1986 international units/ml/10(6) infected cells.

摘要

将编码人尿激酶型纤溶酶原激活剂的cDNA插入杆状病毒苜蓿银纹夜蛾核型多角体病毒多角体蛋白启动子的下游。合成了一种与尿激酶（UK）分子量相似的蛋白质，约90%从重组病毒感染的草地贪夜蛾细胞中分泌出来。对昆虫来源的蛋白质进行酶谱分析和蛋白质免疫印迹分析表明，它仅由高分子量形式的UK组成。未检测到低分子量的UK。酰胺分解活性测定表明，昆虫细胞来源的UK中有96%是单链酶原形式。昆虫细胞中UK的产量为1986国际单位/毫升/10⁶个感染细胞。