Enzymatic characterization of purified NS3 serine proteinase of hepatitis C virus expressed in Escherichia coli

FEBS Letters

Koide, Tomoko¹  Yuasa, Satoshi¹  Moria, Akiko²  Miyamura, Tatsuo³  Yamada, Kazunori²  Yamadaa, Ei²  Kimura, Junko² 

[1] Laboratory of Pharmaceuticals II, Yokohama Research Center, Mitsubishi Chemical Corp., 1000 Kamoshida-cho, Aoba-ku, Yokohama 227, Japan;Laboratory of Molecular Medicine, Yokohama Research Center, Mitsubishi Chemical Corp., 1000 Kamoshida-cho, Aoba-ku, Yokohama 227, Japan;Department of Virology II, National Institute of Health, 1-23-1 Toyama, Shinjuku-ku, Tokyo 162, Japan

关键词: Hepatitis C virus;    Non-structural protein;    Serine proteinase;    Inhibitor;    Chelator;    HCV;    hepatitis C virus;    NS;    non-structural protein;    APMSF;    4-(amidinophenyl)-methanesulfonyl fluoride;    E-64;    l-trans-epoxysuccinyl-leucylamide-(4-guanidino)-butane;    PMSF;    phenylmethanesulfonyl fluoride;    TLCK;    tosyllysylchloromethane;    TPCK;    tosylphenylalanylchloromethane;    Pefabloc/SC;    4-(2-aminoethyl)-benzenesulfonylfluoride;    CyDTA;    trans-1;    2-cyclohexanediamine-N;    N;    N′;    N′-tetraacetic acid;   

DOI  :  10.1016/0014-5793(95)01423-3

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

Non-structural protein 3 (NS3) of the hepatitis C virus (HCV) has been shown to be a serine proteinase which cleaves the HCV polyprotein thus activating its replicative machinery. To characterize enzymatic activities of NS3 serine proteinase, the proteinase region was expressed in Escherichia coli and purified. The purified proteinase specifically cleaved a purified fusion protein sandwiching the NS5A/5B cleavage sequence. In addition to serine proteinase inhibitors, some chelators also inhibited the cleavage activity. Metal ions were not required for its activity, suggesting that the proteinase may be a novel serine proteinase having a unique binding site for chelators.

【 授权许可】

Unknown