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

文献类型: 外文期刊

第一作者: Xu, Ao-Tian

作者: Xu, Ao-Tian;Zhou, Yan-Jun;Li, Guo-Xin;Yu, Hai;Yan, Li-Ping;Tong, Guang-Zhi

作者机构:

关键词: 3C-like proteinase;Nsp4;Porcine reproductive and respiratory syndrome virus

期刊名称:BIOTECHNOLOGY LETTERS ( 影响因子:2.461; 五年影响因子:2.457 )

ISSN: 0141-5492

年卷期: 2010 年 32 卷 12 期

页码:

收录情况: SCI

摘要: Purpose of work 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 degrees C and 7.5, respectively. Na(+) (1000 mM) and K(+) (500 mM) were not inhibitory to its activity but Cu(2+), Zn(2+), PMSF and EDTA were significantly inhibitory. His(39), Asp(64) and Ser(118) residues were identified to form the catalytic triad of PRRSV 3CLpro by a series of site-directed mutagenesis analysis.

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