Construction and characterization of a recombinant chimeric plasminogen   activator consisting of a fibrin peptide and a low molecular mass   single-chain urokinase

doi:10.1016/S0300-9084(01)01352-9

CORC > 北京大学 > 生命科学学院

	Construction and characterization of a recombinant chimeric plasminogen activator consisting of a fibrin peptide and a low molecular mass single-chain urokinase
	Jiao, JW ; Yu, MM ; Ru, BG
刊名	biochimie
	2001
关键词	urokinase fibrin peptide chimeric thrombolysis PRO-UROKINASE PLATELET-AGGREGATION SYNTHETIC PEPTIDES WEIGHT FORM POLYMERIZATION BINDING PROTEIN THERAPY
DOI	10.1016/S0300-9084(01)01352-9
英文摘要	A recombinant chimeric plasminogen activator (fbeta/scuPA-32k), with a fibrin P-chain peptide (comprising Gly 15 through Arg 42) linked to the N-terminal of a low molecular mass (32 kDa) single-chain urokinase (scuPA-32k, comprising Leu144 through Leu 411) via a 50 amino acid linker sequence, was produced by expression the corresponding chimeric cDNA in Escherichia coli cells. After refolding in vitro, the chimeric protein was purified to homogeneity by zinc chelate-Sepharose chromatography, Sephacryl S200 chromatography and benzamidine-Sepharose chromatography in sequence. The apparent molecular mass was 36 kDa shown by SDS-PAGE analysis. The special activity was 87 000 IU/mg detected by fibrin plate determination. Fbeta/scuPA-32k could directly activate plasminogen following Michaelis-Menten kinetics with K-m = 0.52 muM and k(2) = 0.0024 s(-1). Mediated by plasmin, the single-chain molecule could be converted to the active two-chain molecule. The chimeric protein had 3.3 times higher fibrin affinity than scuPA-32k in the fibrin concentration of 3.2 mg/mL, while the chimeric protein inhibited the fibrin clotting and platelet aggregation. Fbeta/scuPA-32k showed a higher thrombolytic potency in vitro plasma clot lysis than scuPA-32k and depleted less fibrinogen in plasma, These results showed that the chimeric protein had not only higher fibrinolytic activity but also anti-thrombus activity. Further evaluation of the thrombolytic potential in appropriate animal models is required. (C) 2001 Societe francaise de biochimie et biologie moleculaire / Editions scientifiques et medicales Elsevier SAS. All rights reserved.; http://gateway.webofknowledge.com/gateway/Gateway.cgi?GWVersion=2&SrcApp=PARTNER_APP&SrcAuth=LinksAMR&KeyUT=WOS:000173495500007&DestLinkType=FullRecord&DestApp=ALL_WOS&UsrCustomerID=8e1609b174ce4e31116a60747a720701 ; Biochemistry & Molecular Biology; SCI(E); 3; ARTICLE; 11-12; 1049-1055; 83
语种	英语
内容类型	期刊论文
源URL	[http://ir.pku.edu.cn/handle/20.500.11897/211037]
专题	生命科学学院
推荐引用方式 GB/T 7714	Jiao, JW,Yu, MM,Ru, BG. Construction and characterization of a recombinant chimeric plasminogen activator consisting of a fibrin peptide and a low molecular mass single-chain urokinase[J]. biochimie,2001.
APA	Jiao, JW,Yu, MM,&Ru, BG.(2001).Construction and characterization of a recombinant chimeric plasminogen activator consisting of a fibrin peptide and a low molecular mass single-chain urokinase.biochimie.
MLA	Jiao, JW,et al."Construction and characterization of a recombinant chimeric plasminogen activator consisting of a fibrin peptide and a low molecular mass single-chain urokinase".biochimie (2001).