南京大学学报（自然科学版）2004，Vol.40Issue(5)：566-575,10.
丝氨酸蛋白酶抑制剂可抑制昆虫细胞表达的蛋白质降解
Serine Protease Inhibitor Inhibits Degradation of the Proteins Expressed in Insect Cells
摘要
Abstract
Stem cell factor (SCF) is a hematopoietic cytokine that promotes the survival,proliferation, and differentiation of hematopoietic cells. The recombinant dual human stem cell factor (rdhSCF) possesses higher specific activity than the monomer form of hSCF and might be preferred since it could be administered at low doses to avoid significant mast cell activation while stimulating hematopoietic recovery. In this report, degradation was found when rdhSCF was expressed in Sf9 cells and E. coli cells. The deletion analysis of rdhSCF confirmed that the cleavage site locates in the region from 145 to 165 amino acid of rdhSCF. Only serine protease inhibitor aprotinin and PMSF could inhibit the site-specific degradation. The influences of aprotinin and PMSF on Sf9 cell survivals and rdhSCF yield were measured at different concentrations. When aprotinin concentration was 1.0 μg/mL, the highest expression level of rdhSCF was achieved, which was about two times higher than without adding protease inhibitor. Aprotinin could completely inhibit degradation of rdhSCF expressed in insect cells.关键词
重组人二连体干细胞因子,杆状病毒表达系统,Sf9细胞,丝氨酸蛋白酶抑制剂,aprotinin,PMSFKey words
rdhSCF, baculovirus expression system, Sf9 cell, Serine protease inhibitor,aprotinin, PMSF分类
生物科学引用本文复制引用
韩俊海,吕海芹,臧宇辉,朱洁,迮玉官,陈涛,秦浚川..丝氨酸蛋白酶抑制剂可抑制昆虫细胞表达的蛋白质降解[J].南京大学学报（自然科学版）,2004,40(5):566-575,10.基金项目
National Natural Science Foundation of China (30270703) （30270703）
