溶栓与抗栓双功能尿激酶原突变体的模拟、构建与表达

doi:10.3321/j.issn:0251-0790.2004.06.007

CORC > 北京大学 > 生命科学学院

	溶栓与抗栓双功能尿激酶原突变体的模拟、构建与表达; Simulation, construction and expression of the recombinant dscuPA with thrombolytic and antithrombus bifunction
	张磊亮 ; 焦建伟 ; 邵开峰 ; 俞梅敏 ; 茹炳根
刊名	高等学校化学学报
	2004
关键词	低分子量尿激酶原抗栓肽血小板聚集 ScuPA-32k decorsin platelet aggregation
DOI	10.3321/j.issn:0251-0790.2004.06.007
英文摘要	将抗栓肽 ( Decorsin)嫁接到低分子量尿激酶原 ( scu PA-3 2 k)上 ,可以期望获得既具有抗血小板聚集活性 ,又具有溶栓活性的新型基因工程蛋白质分子 .利用计算机辅助分子设计手段模拟了该融合蛋白的分子结构 ,证明其活性区可以正常发挥功能 .根据大肠杆菌偏好密码子合成 Decorsin的基因 ,与 scu PA-3 2 k基因融合在一起 ,构建新的嵌合体基因 dscu PA,并在大肠杆菌中通过 IPTG进行诱导表达 ,该重组蛋白在大肠杆菌中以包涵体的形式存在 .对包涵体进行变性和复性并通过层析纯化得到目的蛋白质 .用纤维蛋白平板法测得重组蛋白的比活为 92 0 0 0 IU/...; A recombinant chimeric plasminogen activator (dscuPA) was constructed, consisting of the decorsin (platelet aggregation inhibitor) and a low molecular mass(32 000) single-chain urokinase(scuPA-32k, comprising Leu144 through Leu411). The structure of the designed protein was predicted and simulated. The recombinant protein was produced in E. Coll after IPTG induction and exited in inclusion body. After refolded in vitro, the chimeric protein was purified by chromatography. The special activity of the chimera was 92 000 IU/mg detected by fibrin plate determination. The chimera could activate plasminogen following Michaelis-Menten kinetics with K-m = 1.36 mumol/L and k(cat) = 0.0028 s(-1), corresponding to that of scuPA. It was also shown that chimera inhibited ADP-induced platelet aggregation in a concentration dependent manner. These results showed that the chimeric protein not only had high thrombolytic activity but also had anti-thrombus function.; SCI(E); 中文核心期刊要目总览(PKU); 中国科技核心期刊(ISTIC); 中国科学引文数据库(CSCD); 2; 06; 1046-1050; 25
语种	中文
内容类型	期刊论文
源URL	[http://ir.pku.edu.cn/handle/20.500.11897/46967]
专题	生命科学学院
推荐引用方式 GB/T 7714	张磊亮,焦建伟,邵开峰,等. 溶栓与抗栓双功能尿激酶原突变体的模拟、构建与表达, Simulation, construction and expression of the recombinant dscuPA with thrombolytic and antithrombus bifunction[J]. 高等学校化学学报,2004.
APA	张磊亮,焦建伟,邵开峰,俞梅敏,&茹炳根.(2004).溶栓与抗栓双功能尿激酶原突变体的模拟、构建与表达.高等学校化学学报.
MLA	张磊亮,et al."溶栓与抗栓双功能尿激酶原突变体的模拟、构建与表达".高等学校化学学报 (2004).