Characterization of the norovirus 3C-like protease

被引：23

作者：

Someya, Y ^{[1
]}

Takeda, N ^{[1
]}

Miyamura, T ^{[1
]}

机构：

[1] Natl Inst Infect Dis, Dept Virol 2, Shinjuku Ku, Tokyo 1628640, Japan

来源：

VIRUS RESEARCH | 2005年 / 110卷 / 1-2期

关键词：

3C-like protease; Chiba virus; norovirus; SH reagents; proteolytic activity;

D O I：

10.1016/j.virusres.2005.02.002

中图分类号：

Q93 [微生物学];

学科分类号：

071005 ; 100705 ;

摘要：

The recombinant 3C-fike protease of Chiba virus, a Norovirus, expressed in Escherichia coli cells was purified and characterized as to effects of pH, temperature, salt contents, and SH reagents on its proteolytic activity. The optimal pH and temperature of the 3C-like protease for the proteolytic activity were 8.6 and 37 degrees C, respectively. Increased concentration (similar to 100mM) of monovalent cations such as Na+ and K+ was inhibitory to the activity. Hg2+ and Zn2+ remarkably inhibited the protease activity, while Mg2+ and Ca2+ had no virtual effect. Several sulthydryl reagents such as p-chloromercuribenzoic acid, methyl methanethiosulfonate, N-ethylmaleimide and N-phenylmaleimide also blocked the activity, confirming the previous result that cysteine residue(s) were responsible for the proteolysis. (c) 2005 Elsevier B.V. All rights reserved.

引用

页码：91 / 97

页数：7

共 27 条

[1] Coronavirus main proteinase (3CLpro) structure:: Basis for design of anti-SARS drugs [J].

Anand, K ;

Ziebuhr, J ;

Wadhwani, P ;

Mesters, JR ;

Hilgenfeld, R .

SCIENCE, 2003, 300 (5626) :1763-1767

[2] Structure of coronavirus main proteinase reveals combination of a chymotrypsin fold with an extra α-helical domain [J].

Anand, K ;

Palm, GJ ;

Mesters, JR ;

Siddell, SG ;

Ziebuhr, J ;

Hilgenfeld, R .

EMBO JOURNAL, 2002, 21 (13) :3213-3224

[3]

[Anonymous], 1997, INT J ED TELECOMMUNI

[4]

Barrett JW, 2002, INTERFACE FREE BOUND, V4, P277

[5] PURIFICATION, PROPERTIES, AND MUTAGENESIS OF POLIOVIRUS 3C PROTEASE [J].

BAUM, EZ ;

BEBERNITZ, GA ;

PALANT, O ;

MUELLER, T ;

PLOTCH, SJ .

VIROLOGY, 1991, 185 (01) :140-150

[6] In vitro proteolytic processing of the MD145 norovirus ORF1 nonstructural polyprotein yields stable precursors and products similar to those detected in calicivirus-infected cells [J].

Belliot, G ;

Sosnovtsev, SV ;

Mitra, T ;

Hammer, C ;

Garfield, M ;

Green, KY .

JOURNAL OF VIROLOGY, 2003, 77 (20) :10957-10974

[7] IDENTIFICATION AND CHARACTERIZATION OF A 3C-LIKE PROTEASE FROM RABBIT HEMORRHAGIC-DISEASE VIRUS, A CALICIVIRUS [J].

BONIOTTI, B ;

WIRBLICH, C ;

SIBILIA, M ;

MEYERS, G ;

THIEL, HJ ;

ROSSI, C .

JOURNAL OF VIROLOGY, 1994, 68 (10) :6487-6495

[8]

BRADFORD MM, 1976, ANAL BIOCHEM, V72, P248, DOI 10.1016/0003-2697(76)90527-3

[9]

CHEAH KC, 1990, J BIOL CHEM, V265, P7180

[10] Expression and partial purification of recombinant tomato ringspot nepovirus 3C-like proteinase:: comparison of the activity of the mature proteinase and the VPg-proteinase precursor [J].

Chisholm, J ;

Wieczorek, A ;

Sanfaçon, H .

VIRUS RESEARCH, 2001, 79 (1-2) :153-164

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