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Substrate Binding and Homo-Dimerization of SARS 3CL Proteinase are Mutual Allosteric Effectors
Wei Ping1,2; Li Chun-Mei1,2; Zhou Lu1; Liu Ying1; Lai Lu-Hua1,2
2010-04-01
Source PublicationACTA PHYSICO-CHIMICA SINICA
ISSN1000-6818
Volume26Issue:4Pages:1093-1098
AbstractThe 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
KeywordAllosteric Effect Sars 3cl Proteinase Analytical Ultracentrifugation Substrate Enhanced Enzyme Dimenzation
DOI10.3866/PKU.WHXB20100449
Indexed BySCI
Language英语
WOS IDWOS:000276958400038
PublisherPEKING UNIV PRESS
Citation statistics
Cited Times:3[WOS]   [WOS Record]     [Related Records in WOS]
Document Type期刊论文
Identifierhttp://ir.iccas.ac.cn/handle/121111/69168
Collection中国科学院化学研究所
Corresponding AuthorLai Lu-Hua
Affiliation1.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
Recommended Citation
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.
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