Purification and characterization of the NS3 serine protease domain of hepatitis C virus expressed in Saccharomyces cerevisiae

被引：0

作者：

Markland, W ^{[1
]}

Petrillo, RA ^{[1
]}

Fitzgibbon, M ^{[1
]}

Fox, T ^{[1
]}

McCarrick, R ^{[1
]}

McQuaid, T ^{[1
]}

Fulghum, JR ^{[1
]}

Chen, WY ^{[1
]}

Fleming, MA ^{[1
]}

Thomson, JA ^{[1
]}

Chambers, SP ^{[1
]}

机构：

[1] VERTEX PHARMACEUT INC, CAMBRIDGE, MA 02139 USA

来源：

JOURNAL OF GENERAL VIROLOGY | 1997年 / 78卷

关键词：

D O I：

暂无

中图分类号：

Q81 [生物工程学（生物技术）]; Q93 [微生物学];

学科分类号：

071005 ; 0836 ; 090102 ; 100705 ;

摘要：

cDNA encoding the putative core of the hepatitis C virus NS3 serine protease domain (residues 1-181 of NS3; NS3(181)) was expressed as an N-terminally (His)(6)-tagged fusion protein in Saccharomyces cerevisiae. NS3(181) protease activity was found in soluble cell lysates, and the N-terminal metal-chelating domain facilitated the efficient purification of active enzyme, using immobilized metal affinity chromatography. The purified NS3(181) protease activity was characterized by assaying the trans-cleavage of in vitro transcription-translation generated substrates, and subsequently a previously unobserved cleavage site within the NS5A region was identified. The inhibitory effect of known protease inhibitors was also examined. It is hoped that availability of this method for the expression and purification of the NS3(181) protease will facilitate the development of anti-hepatitis C therapies.

引用

页码：39 / 43

页数：5

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