A novel auto-cleavage assay for studying mutational effects on the active site of severe acute respiratory syndrome coronavirus 3C-like protease
文献类型：期刊论文
| 作者 | Shan, YF; Li, SF; Xu, GJ |
| 刊名 | BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS
 |
| 出版日期 | 2004 |
| 卷号 | 324期号:2页码:579-583 |
| 关键词 | SARS 3CL protease active site auto-cleavage assay N-31-tag |
| 通讯作者 | Xu, GJ (reprint author), Chinese Acad Sci, Shanghai Inst Biol Sci, Inst Biochem & Cell Biol, Shanghai 200031, Peoples R China.,gjxu@sibs.ac.cn |
| 英文摘要 | The 3C-like protease (3CL(pro)) of severe acute respiratory syndrome (SARS) has been proposed as an attractive target for drug design. His(41) and Cys(145) were essential for the active site as the principal catalytic residues. In this study, we mutated the two sites, expressed four resulting mutants in Escherichia coli and characterized. All mutants showed undetectable activity in trans-cleavage assay. In addition. we introduced a 31-mer peptide containing an auto-cleavage site to the N-terminal of the proteases and found the peptide could be cleaved efficiently by 3CLsc itself, but, among the four mutants, only the mutant cys(145) --> Ser showed residual activity as detected by the auto-cleavage assay. The data supported the proposition unequivocally that SARS-CoV 3CL(pro) was a member of serine proteases involving His(41) and Cys(145) residues at the active site. The auto-cleavage assay also provided a sensitive and reliable compensation to the traditional trans-cleavage assay. (C) 2004 Elsevier Inc. All rights reserved. |
| 学科主题 | Biochemistry & Molecular Biology; Biophysics |
| 类目[WOS] | Biochemistry & Molecular Biology ; Biophysics |
| 关键词[WOS] | SARS CORONAVIRUS ; PROTEINASE ; VIRUS ; PURIFICATION ; BIOSYNTHESIS ; DESIGN ; DOMAIN |
| 收录类别 | SCI |
| 语种 | 英语 |
| WOS记录号 | WOS:000224794000016 |
| 版本 | 出版稿 |
| 源URL | [http://202.127.25.143/handle/331003/2030]  |
| 专题 | 上海生化细胞研究所_上海生科院生化细胞研究所 |
| 推荐引用方式 GB/T 7714 | Shan, YF,Li, SF,Xu, GJ. A novel auto-cleavage assay for studying mutational effects on the active site of severe acute respiratory syndrome coronavirus 3C-like protease[J]. BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS,2004,324(2):579-583. |
| APA | Shan, YF,Li, SF,&Xu, GJ.(2004).A novel auto-cleavage assay for studying mutational effects on the active site of severe acute respiratory syndrome coronavirus 3C-like protease.BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS,324(2),579-583. |
| MLA | Shan, YF,et al."A novel auto-cleavage assay for studying mutational effects on the active site of severe acute respiratory syndrome coronavirus 3C-like protease".BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS 324.2(2004):579-583. |
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