Structure and cleavage specificity of the chymotrypsin-like serine protease (3clsp/nsp4) of porcine reproductive and respiratory syndrome virus (prrsv)
文献类型：期刊论文
| 作者 | Tian, Xinsheng1,2; Lu, Guangwen1,2; Gao, Feng1,3; Peng, Hao1; Feng, Youjun1,2; Ma, Guangpeng1,4; Bartlam, Mark5,6; Tian, Kegong7; Yan, Jinghua1; Hilgenfeld, Rolf8 |
| 刊名 | Journal of molecular biology
 |
| 出版日期 | 2009-10-02 |
| 卷号 | 392期号:4页码:977-993 |
| 关键词 | Prrsv 3c-like serine protease Structure Trans/cis cleavage |
| ISSN号 | 0022-2836 |
| DOI | 10.1016/j.jmb.2009.07.062 |
| 通讯作者 | Gao, george f.(gaof@im.ac.cn) |
| 英文摘要 | Biogenesis and replication of the porcine reproductive and respiratory syndrome virus (prrsv) include the crucial step of replicative polyprotein processing by self-encoded proteases. whole genome bioinformatics analysis suggests that nonstructural protein 4 (nsp4) is a 3c-like serine protease (3clsp), responsible for most of the nonstructural protein processing. the gene encoding this protease was cloned and expressed in escherichia coli in order to confirm this prediction. the purified protein was crystallized, and the structure was solved at 1.9 angstrom resolution. in addition, the crystal structure of the ser118ala mutant was determined at 2.0 angstrom resolution. the monomeric enzyme folds into three domains, similar to that of the homologous protease of equine arteritis virus, which, like prrsv, is a member of the family arteriviridae in the order of nidovirales. the active site of the prrsv 3clsp is located between domains i and 11 and harbors a canonical catalytic triad comprising ser118, hs39, and asp64. the structure also shows an atypical oxyanion hole and a partially collapsed s1 specificity pocket. the proteolytic activity of the purified protein was assessed in vitro. three sites joining nonstructural protein domains in the prrsv replicative polyprotein are confirmed to be processed by the enzyme. two of them, the nsp3/nsp4 and nsp11/nsp12 junctions, are shown to be cleaved in trans, while cis cleavage is demonstrated for the nsp4/nsp5 linker. thus, we provide structural evidence as well as enzymatic proof of the nsp4 protein being a functional 3clsp. we also show that the enzyme has a strong preference for glutamic acid at the p1 position of the substrate. (c) 2009 published by elsevier ltd. |
| WOS关键词 | CORONAVIRUS MAIN PROTEINASE ; RHINOVIRUS 3C PROTEASE ; ARTERIVIRUS REPLICASE ; 3C-LIKE PROTEINASE ; CRYSTAL-STRUCTURE ; UNITED-STATES ; GENE-PRODUCT ; EXTRA DOMAIN ; NSP4 ; PURIFICATION |
| WOS研究方向 | Biochemistry & Molecular Biology |
| WOS类目 | Biochemistry & Molecular Biology |
| 语种 | 英语 |
| WOS记录号 | WOS:000270601200012 |
| 出版者 | ACADEMIC PRESS LTD- ELSEVIER SCIENCE LTD |
| URI标识 | http://www.irgrid.ac.cn/handle/1471x/2404217 |
| 专题 | 中国科学院大学 |
| 通讯作者 | Gao, George F. |
| 作者单位 | 1.Chinese Acad Sci, CAS Key Lab Pathogen Microbiol & Immunol, Inst Microbiol, Beijing 100101, Peoples R China 2.Chinese Acad Sci, Grad Univ, Beijing 100049, Peoples R China 3.China Agr Univ, Coll Life Sci, Beijing 100094, Peoples R China 4.NE Agr Univ, Coll Vet Med, Dept Vet Prevent Med, Harbin 150030, Peoples R China 5.Nankai Univ, Coll Life Sci, Tianjin 300071, Peoples R China 6.Nankai Univ, Tianjin Key Lab Prot Sci, Tianjin 300071, Peoples R China 7.China Anim Dis Control Ctr, Beijing 100094, Peoples R China 8.Med Univ Lubeck, Inst Biochem, Ctr Struct & Cell Biol Med, D-23538 Lubeck, Germany 9.Chinese Acad Sci, Inst Microbiol, China Japan Joint Lab Mol Immunol & Mol Microbiol, Beijing 100101, Peoples R China 10.Chinese Acad Sci, Beijing Inst Life Sci, Beijing 100101, Peoples R China |
| 推荐引用方式 GB/T 7714 | Tian, Xinsheng,Lu, Guangwen,Gao, Feng,et al. Structure and cleavage specificity of the chymotrypsin-like serine protease (3clsp/nsp4) of porcine reproductive and respiratory syndrome virus (prrsv)[J]. Journal of molecular biology,2009,392(4):977-993. |
| APA | Tian, Xinsheng.,Lu, Guangwen.,Gao, Feng.,Peng, Hao.,Feng, Youjun.,...&Gao, George F..(2009).Structure and cleavage specificity of the chymotrypsin-like serine protease (3clsp/nsp4) of porcine reproductive and respiratory syndrome virus (prrsv).Journal of molecular biology,392(4),977-993. |
| MLA | Tian, Xinsheng,et al."Structure and cleavage specificity of the chymotrypsin-like serine protease (3clsp/nsp4) of porcine reproductive and respiratory syndrome virus (prrsv)".Journal of molecular biology 392.4(2009):977-993. |
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来源：中国科学院大学
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