Expression and characterization of ARSP1 from Eisenia fetida

doi:10.1016/j.cca.2003.12.003

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	Expression and characterization of ARSP1 from Eisenia fetida
	Zhang, LL ; Wang, J ; Yu, MM ; Ru, BG
刊名	comparative biochemistry and physiology c toxicology pharmacology
	2004
关键词	apoptosis-related serine protease serine protease fibrinolytic enzyme alignment fibrinogen plasminogen thrombin platelet aggregation FIBRINOLYTIC ENZYME BACTERIAL EXPRESSION LUMBRICUS-RUBELLUS PHOSPHOLIPASE A(2) SERINE-PROTEASE EARTHWORM PURIFICATION THERAPY CLONING
DOI	10.1016/j.cca.2003.12.003
英文摘要	To study the characterization of a protease ARSP1 (apoptosis-related serine protease) of Eisenia fetida, a recombinant ARSP1 was constructed. ARSP1 was produced in E. coli BL21-CodonPlus (DE3)-RIL after IPTG induction and exited in inclusion body. After refolding in vitro, the protein was purified by DEAE-Sepharose F.F. and Sephacryl S-100 chromatography in sequence. ARSP1 showed high sequence identity to other chymotrypsin-like serine proteases and the catalytic triad was His(41)-Asp(90)-Ser(188). ARSP1 could degrade casein following Michaelis-Menten kinetics with a V-max of 43.9 U/mg protein and a K-m for casein of 0.83 g/l. Studies with inhibitors indicated that ARSP1 was a chymotrypsin-like serine protease. Experiments in vitro demonstrated that ARSP1 could not only hydrolyze fibrinogen and fibrin directly, but also activate plasminogen to plasmin. ARSP1 inhibited thrombin activity and ADP-induced platelet aggregation in a dose-response correlation. These results showed that ARSP1 has thrombolytic activity and also an anti-thrombus function. (C) 2004 Elsevier Inc. All rights reserved.; http://gateway.webofknowledge.com/gateway/Gateway.cgi?GWVersion=2&SrcApp=PARTNER_APP&SrcAuth=LinksAMR&KeyUT=WOS:000220637700003&DestLinkType=FullRecord&DestApp=ALL_WOS&UsrCustomerID=8e1609b174ce4e31116a60747a720701 ; Biochemistry & Molecular Biology; Endocrinology & Metabolism; Toxicology; Zoology; SCI(E); PubMed; 1; ARTICLE; 2; 115-122; 137
语种	英语
内容类型	期刊论文
源URL	[http://ir.pku.edu.cn/handle/20.500.11897/199611]
专题	生命科学学院
推荐引用方式 GB/T 7714	Zhang, LL,Wang, J,Yu, MM,et al. Expression and characterization of ARSP1 from Eisenia fetida[J]. comparative biochemistry and physiology c toxicology pharmacology,2004.
APA	Zhang, LL,Wang, J,Yu, MM,&Ru, BG.(2004).Expression and characterization of ARSP1 from Eisenia fetida.comparative biochemistry and physiology c toxicology pharmacology.
MLA	Zhang, LL,et al."Expression and characterization of ARSP1 from Eisenia fetida".comparative biochemistry and physiology c toxicology pharmacology (2004).