Substrate Binding and Homo-Dimerization of SARS 3CL Proteinase are Mutual Allosteric Effectors
文献类型：期刊论文
| 作者 | Wei Ping1,2; Li Chun-Mei1,2; Zhou Lu1; Liu Ying1; Lai Lu-Hua1,2 |
| 刊名 | ACTA PHYSICO-CHIMICA SINICA
 |
| 出版日期 | 2010-04-01 |
| 卷号 | 26期号:4页码:1093-1098 |
| 关键词 | Allosteric Effect Sars 3cl Proteinase Analytical Ultracentrifugation Substrate Enhanced Enzyme Dimenzation |
| ISSN号 | 1000-6818 |
| DOI | 10.3866/PKU.WHXB20100449 |
| 英文摘要 | The 3C-like proteinase (3CLpro) of severe acute respiratory syndrome (SARS) coronavirus has been proposed to be a key target for anti-SARS drug discovery. It has been proposed and verified that the dimer was the active form of 3CLpro and only one protomer is active In our previous work, we measured the dissociation constant (K-d) of the purified SARS 3CLpro using analytical ultracentrifugation at around 14.0 mu mol . L-1 Using this K-d value, most of the SARS 3CLpro in the in vitro activity assay (1-3 mu m . L-1) might be in the monomer form and inactive. To explain this dilemma, we measured the enzyme activity change together with the enzyme concentration. By fitting the concentration dependent activity profile, the apparent dissociation constant was found to be 094 mu mol . L-1. Indicating a clear tendency toward substrate enhanced dimerization. This also explains why SARS 3CLpro was still active in the in vitro activity assay under a relatively low enzyme concentration. To further verify the substrate induced dimerization phenomenon, we selected a previously reported SARS 3CLpro isatin inhibitor, 1-(2-naphthlmethyl) isatin-5-carboxamide (5f), which has similar binding interactions with the substrate and we studied its influence on SARS 3CLpro dimer formation using analytical ultracentrifugation 5f showed a strong ability to Induce SARS 3CLpro dimer formation By measuring the dimer and monomer distribution under different 5f concentrations, the EC50 of dirtier induction was found to be about 1.0 mu mol . L-1 under an enzyme concentration of 3 0 mu mol . L-1. This implies that only one protomer in the SARS 3CLpro choler binds to the inhibitor or the substrate. As the apparent association constant and thus the enzyme activity of SARS 3CLpro increases with the concentration of the substrate, this may be a smart way to allostetrically regulate the hydrolysis of the SARS viral polyproteins and the correct assembly of virons |
| 语种 | 英语 |
| WOS记录号 | WOS:000276958400038 |
| 出版者 | PEKING UNIV PRESS |
| 源URL | [http://ir.iccas.ac.cn/handle/121111/69167]  |
| 专题 | 中国科学院化学研究所 |
| 通讯作者 | Lai Lu-Hua |
| 作者单位 | 1.Peking Univ, Coll Chem & Mol Engn, State Key Lab Struct Chem Unstable & Stable Speci, Beijing Natl Lab Mol Sci, Beijing 100871, Peoples R China 2.Peking Univ, Ctr Theoret Biol, Beijing 100871, Peoples R China |
| 推荐引用方式 GB/T 7714 | Wei Ping,Li Chun-Mei,Zhou Lu,et al. Substrate Binding and Homo-Dimerization of SARS 3CL Proteinase are Mutual Allosteric Effectors[J]. ACTA PHYSICO-CHIMICA SINICA,2010,26(4):1093-1098. |
| APA | Wei Ping,Li Chun-Mei,Zhou Lu,Liu Ying,&Lai Lu-Hua.(2010).Substrate Binding and Homo-Dimerization of SARS 3CL Proteinase are Mutual Allosteric Effectors.ACTA PHYSICO-CHIMICA SINICA,26(4),1093-1098. |
| MLA | Wei Ping,et al."Substrate Binding and Homo-Dimerization of SARS 3CL Proteinase are Mutual Allosteric Effectors".ACTA PHYSICO-CHIMICA SINICA 26.4(2010):1093-1098. |
入库方式： OAI收割
来源：化学研究所
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