Characterization of the biochemical properties and identification of amino acids forming the catalytic center of 3C-like proteinase of porcine reproductive and respiratory syndrome virus.
Characterization of the biochemical properties and identification of amino acids forming the catalytic center of 3C-like proteinase of porcine reproductive and respiratory syndrome virus.
复制标题
DOI：
10.1007/s10529-010-0370-1
复制
发表时间：
2010-12
影响因子：
2.7
通讯作者：
Tong GZ
中科院分区：
文献类型：
--
作者：
Xu AT;Zhou YJ;Li GX;Yu H;Yan LP;Tong GZ
猪繁殖与呼吸综合征病毒（PRRSV）非结构蛋白4（Nsp 4）是一种3C样蛋白酶（3CLpro），在基因表达和复制中起着关键作用。我们已经研究了PRRSV 3CLpro的生化特性，并确定了参与其催化活性的氨基酸残基作为开发抗PRRSV策略的前奏。在大肠杆菌中表达了猪繁殖与呼吸综合征病毒（PRRSV）3C样蛋白酶（3CLpro），并对其进行了鉴定。其蛋白水解活性的最适温度为8°C，最适pH为7.5。Na+（1000 mM）和K+（500 mM）对该酶活性无抑制作用，而Cu ~（2+）、Zn ~（2+）、PMSF和EDTA对该酶活性有显著抑制作用。通过一系列的定点突变分析，确定His 39、Asp 64和Ser 118残基形成PRRSV 3CLpro的催化三联体。本文的在线版本（doi：10.1007/s10529-010-0370-1）包含补充材料，可供授权用户使用。
The non-structural protein 4 (Nsp4) of porcine reproductive and respiratory syndrome virus (PRRSV) functions as a 3C-like proteinase (3CLpro) and plays a pivotal role in gene expression and replication. We have examined the biochemical properties of PRRSV 3CLpro and identified those amino acid residues involved in its catalytic activity as a prelude to developing anti-PRRSV strategies. The 3C-like proteinase (3CLpro) of porcine reproductive and respiratory syndrome virus (PRRSV) was expressed in Escherichia coli and characterized. The optimal temperature and pH for its proteolytic activity were 8°C and 7.5, respectively. Na+ (1000 mM) and K+ (500 mM) were not inhibitory to its activity but Cu2+, Zn2+, PMSF and EDTA were significantly inhibitory. His39, Asp64 and Ser118 residues were identified to form the catalytic triad of PRRSV 3CLpro by a series of site-directed mutagenesis analysis. The online version of this article (doi:10.1007/s10529-010-0370-1) contains supplementary material, which is available to authorized users.
登录
查看更多内容
影响因子：
2.7
作者：
Fisher, CL;Pei, GK
通讯作者：
Pei, GK
影响因子：
5
作者：
van Aken D;Benckhuijsen WE;Drijfhout JW;Wassenaar AL;Gorbalenya AE;Snijder EJ
通讯作者：
Snijder EJ
DOI：
10.2460/javma.2005.227.385
发表时间：
2005-08-01
影响因子：
1.9
作者：
Neumann, EJ;Kliebenstein, JB;Zimmerman, JJ
通讯作者：
Zimmerman, JJ
影响因子：
4.8
作者：
Snijder, EJ;Wassenaar, ALM;Gorbalenya, AE
通讯作者：
Gorbalenya, AE
牛乳糖蛋白酶样丝氨酸蛋白酶（3clsp/nsp4）生殖和呼吸综合征病毒（PRRSV）的结构和切割特异性。
影响因子：
5.6
作者：
Tian X;Lu G;Gao F;Peng H;Feng Y;Ma G;Bartlam M;Tian K;Yan J;Hilgenfeld R;Gao GF
通讯作者：
Gao GF