enzymaticactivitycharacterizationofsarscoronavirus3clikeproteasebyfluorescenceresonanceenergytransfertechnique1
文献类型：期刊论文
| 作者 | Shuai CHEN; Lili CHEN ; Haibin LUO; Jing CHEN; Fei YE; Jianhua CAI; Jingkang SHEN; Xu SHEN
|
| 刊名 | actapharmacologicasinica
 |
| 出版日期 | 2005 |
| 卷号 | 26期号:1页码:99 |
| 关键词 | severe acute respiratory coronavirus 3C-like protease fluorescence resonance energy transfer fluorogenic substrate enzyme activity site-directed mutagenesis |
| ISSN号 | 1671-4083 |
| 英文摘要 | To characterize enzymatic activity of severe acute respiratory syndrome (SARS) coronavirus (CoV) 3C-like protease (3CLpro) and its four site-directed mutants. Methods: Based on the fluorescence resonance energy transfer (FRET) principle using 5-(2′-aminoethyl)-amino naphthelenesulfonic acid (EDANS) and 4-4-(dimethylamino) phenyl azo benzoic acid (Dabcyl) as the energy transfer pair, one fluorogenic substrate was designed for the evaluation of SARS-CoV 3CLpro proteolytic activity. Results: The kinetic parameters of the fluorogenic substrate have been determined as Km=404 mmol·L-1, kcat=1.08 min-1, and kcat/Km=2.7 mmol-1·L·min-1. SARS-CoV 3CLpro showed substantial pH and temperature-triggered activity switches, and site-directed mutagenesis analysis of SARS-CoV 3CLpro revealed that substitutions of His41, Cys145, and His163 resulted in complete loss of enzymatic activity, while replacement of Met162 with Ala caused strongly increased activity. Conclusion: This present work has provided valuable information for understanding the catalytic mechanism of SARS-CoV 3CLpro. This FRET-based assay might supply an ideal approach for the exploration SARS-CoV 3CLpro putative inhibitors. |
| 语种 | 英语 |
| 源URL | [http://119.78.100.183/handle/2S10ELR8/286049]  |
| 专题 | 中国科学院上海药物研究所 |
| 作者单位 | 中国科学院上海药物研究所 |
| 推荐引用方式 GB/T 7714 | Shuai CHEN,Lili CHEN,Haibin LUO,et al. enzymaticactivitycharacterizationofsarscoronavirus3clikeproteasebyfluorescenceresonanceenergytransfertechnique1[J]. actapharmacologicasinica,2005,26(1):99. |
| APA | Shuai CHEN.,Lili CHEN.,Haibin LUO.,Jing CHEN.,Fei YE.,...&Xu SHEN.(2005).enzymaticactivitycharacterizationofsarscoronavirus3clikeproteasebyfluorescenceresonanceenergytransfertechnique1.actapharmacologicasinica,26(1),99. |
| MLA | Shuai CHEN,et al."enzymaticactivitycharacterizationofsarscoronavirus3clikeproteasebyfluorescenceresonanceenergytransfertechnique1".actapharmacologicasinica 26.1(2005):99. |
入库方式： OAI收割
来源：上海药物研究所
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