PURIFICATION AND CHARACTERIZATION OF HUMAN PLASMINOGEN-ACTIVATOR INHIBITOR TYPE-I EXPRESSED IN SACCHAROMYCES-CEREVISIAE

被引：21

作者：

GARDELL, SJ ^{[1
]}

HARE, TR ^{[1
]}

HAN, JH ^{[1
]}

MARKUS, HZ ^{[1
]}

KEECH, BJ ^{[1
]}

CARTY, CE ^{[1
]}

ELLIS, RW ^{[1
]}

SCHULTZ, LD ^{[1
]}

机构：

[1] MERCK SHARP & DOHME LTD,DEPT VIRUS & CELL BIOL,W POINT,PA 19486

来源：

ARCHIVES OF BIOCHEMISTRY AND BIOPHYSICS | 1990年 / 278卷 / 02期

关键词：

D O I：

10.1016/0003-9861(90)90286-8

中图分类号：

Q5 [生物化学]; Q7 [分子生物学];

学科分类号：

071010 ; 081704 ;

摘要：

The rapidly acting inhibitor of plasminogen activators, PAI-1, was produced intracellularly in Saccharomyces cerevisiae by using the ADH2 promoter to drive the expression of the human PAI-1 cDNA. Approximately 8 mg of human PAI-1 was produced per liter of confluent yeast culture. A purification scheme which resulted in 20% recovery of isolated PAI-1 from the broken yeast cell homogenate was devised. Yeast-derived human PAI-1 differs from endothelial-type PAI-1 isolated from HT1080 fibrosarcoma cells in that the recombinant inhibitor does not contain carbohydrate side chains. Nevertheless, the activity and other functional attributes of yeast-derived PAI-1 are similar to those exhibited by HT1080 fibrosarcoma cell-derived PAI-1. Hence, this study demonstrates that expression of human PAI-1 in yeast is a viable strategy for the production of ample quantities of this key modulator of plasminogen activator-mediated proteolysis. © 1990.

引用

页码：467 / 474

页数：8

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