Home

Citation: WEI Ping, LI Chun-Mei, ZHOU Lu, LIU Ying, LAI Lu-Hua. Substrate Binding and Homo-Dimerization of SARS 3CL Proteinase are Mutual Allosteric Effectors[J]. Acta Physico-Chimica Sinica, ;2010, 26(04): 1093-1098. doi: 10.3866/PKU.WHXB20100449 shu

Substrate Binding and Homo-Dimerization of SARS 3CL Proteinase are Mutual Allosteric Effectors

1.
Beijing National Laboratory for Molecular Science, State Key Laboratory of Structural Chemistry of Unstable and Stable Species, College of Chemistry and Molecular Engineering, Peking University, Beijing 100871, P. R. China; Center for Theoretical Biology, Peking University, Beijing 100871, P. R. China

Received Date: 30 November 2009
Available Online: 17 March 2010

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 (Kd) of the purified SARS 3CLpro using analytical ultracentrifugation at around 14.0 μmol·L-1. Using this Kd value, most of the SARS 3CLpro in the in vitro activity assay (1-3 μmol·L-1) might be in the monomer formand 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 0.94 μmol·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 dimer induction was found to be about 1.0 μmol·L-1 under an enzyme concentration of 3.0 μmol·L-1. This implies that only one protomer in the SARS 3CLpro dimer 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 allosterically regulate the hydrolysis of the SARS viral polyproteins and the correct assembly of virons.
- Allosteric effect,
- SARS,
- 3CL proteinase,
- Analytical ultracentrifugation,
- Substrate enhanced enzyme dimerization
1. [1]
  Gengchen Guo , Tianyu Zhao , Ruichang Sun , Mingzhe Song , Hongyu Liu , Sen Wang , Jingwen Li , Jingbin Zeng . Au-Fe₃O₄ dumbbell-like nanoparticles based lateral flow immunoassay for colorimetric and photothermal dual-mode detection of SARS-CoV-2 spike protein. Chinese Chemical Letters, 2024, 35(6): 109198-. doi: 10.1016/j.cclet.2023.109198
2. [2]
  Cheng PENG , Jianwei WEI , Yating CHEN , Nan HU , Hui ZENG . First principles investigation about interference effects of electronic and optical properties of inorganic and lead-free perovskite Cs₃Bi₂X₉ (X=Cl, Br, I). Chinese Journal of Inorganic Chemistry, 2024, 40(3): 555-560. doi: 10.11862/CJIC.20230282
3. [3]
  Tieping CAO , Yuejun LI , Dawei SUN . Surface plasmon resonance effect enhanced photocatalytic CO₂ reduction performance of S-scheme Bi₂S₃/TiO₂ heterojunction. Chinese Journal of Inorganic Chemistry, 2025, 41(5): 903-912. doi: 10.11862/CJIC.20240366
4. [4]
  Yaling Chen . Basic Theory and Competitive Exam Analysis of Dynamic Isotope Effect. University Chemistry, 2024, 39(8): 403-410. doi: 10.3866/PKU.DXHX202311093
5. [5]
  Xingyuan Lu , Yutao Yao , Junjing Gu , Peifeng Su . Energy Decomposition Analysis and Its Application in the Many-Body Effect of Water Clusters. University Chemistry, 2025, 40(3): 100-107. doi: 10.12461/PKU.DXHX202405074
6. [6]
  Weikang Wang , Yadong Wu , Jianjun Zhang , Kai Meng , Jinhe Li , Lele Wang , Qinqin Liu . Green H₂O₂ synthesis via melamine-foam supported S-scheme Cd_0.5Zn_0.5In₂S₄/S-doped carbon nitride heterojunction: synergistic interfacial charge transfer and local photothermal effect. Acta Physico-Chimica Sinica, 2025, 41(8): 100093-0. doi: 10.1016/j.actphy.2025.100093
7. [7]
  Kexin Dong , Chuqi Shen , Ruyu Yan , Yanping Liu , Chunqiang Zhuang , Shijie Li . Integration of Plasmonic Effect and S-Scheme Heterojunction into Ag/Ag₃PO₄/C₃N₅ Photocatalyst for Boosted Photocatalytic Levofloxacin Degradation. Acta Physico-Chimica Sinica, 2024, 40(10): 2310013-0. doi: 10.3866/PKU.WHXB202310013
8. [8]
  Liwei Wang , Guangran Ma , Li Wang , Fugang Xu . A Comprehensive Analytical Chemistry Experiment: Colorimetric Detection of Vitamin C Using Nanozyme and Smartphone. University Chemistry, 2024, 39(8): 255-262. doi: 10.3866/PKU.DXHX202312094
9. [9]
  Zhuoya WANG , Le HE , Zhiquan LIN , Yingxi WANG , Ling LI . Multifunctional nanozyme Prussian blue modified copper peroxide: Synthesis and photothermal enhanced catalytic therapy of self-provided hydrogen peroxide. Chinese Journal of Inorganic Chemistry, 2024, 40(12): 2445-2454. doi: 10.11862/CJIC.20240194
10. [10]
  Yunxin Li , Jinghui Zhang , Jisen Chen , Feng Zhu , Zhiqiang Liu , Peng Bao , Wei Shen , Sheng Tang . Detection of SARS-CoV-2 based on artificial intelligence-assisted smartphone: A review. Chinese Chemical Letters, 2024, 35(7): 109220-. doi: 10.1016/j.cclet.2023.109220
11. [11]
  Yang Li , Ning Sheng , Kun Wang , Yuhuan Li , Jiandong Jiang , Jinlan Zhang . Azvudine alleviates SARS-CoV-2-induced inflammation by targeting myeloperoxidase in NETosis. Chinese Chemical Letters, 2025, 36(5): 110238-. doi: 10.1016/j.cclet.2024.110238
12. [12]
  Chunyu Yan , Qinglong Qiao , Wei Zhou , Xuelian Zhou , Yonghui Chen , Lu Miao , Zhaochao Xu . FRET-based in vitro assay for rapid detecting of SARS-CoV-2 entry inhibitors. Chinese Chemical Letters, 2025, 36(5): 110258-. doi: 10.1016/j.cclet.2024.110258
13. [13]
  Yang Li , Jiachen Li , Daidi Fan . 二硫化钼纳米片的制备及其纳米酶性能探究——介绍一个大学化学综合实验. University Chemistry, 2025, 40(8): 233-240. doi: 10.12461/PKU.DXHX202410016
14. [14]
  Mengfei He , Chao Chen , Yue Tang , Si Meng , Zunfa Wang , Liyu Wang , Jiabao Xing , Xinyu Zhang , Jiahui Huang , Jiangbo Lu , Hongmei Jing , Xiangyu Liu , Hua Xu . Epitaxial Growth of Nonlayered 2D MnTe Nanosheets with Thickness-Tunable Conduction for p-Type Field Effect Transistor and Superior Contact Electrode. Acta Physico-Chimica Sinica, 2025, 41(2): 2310029-0. doi: 10.3866/PKU.WHXB202310029
15. [15]
  Yanglin Jiang , Mingqing Chen , Min Liang , Yige Yao , Yan Zhang , Peng Wang , Jianping Zhang . Experimental and Theoretical Investigations of Solvent Polarity Effect on ESIPT Mechanism in 4′-N,N-diethylamino-3-hydroxybenzoflavone. Acta Physico-Chimica Sinica, 2025, 41(2): 2309027-0. doi: 10.3866/PKU.WHXB202309027
16. [16]
  Yihui Song , Shangshang Qin , Kai Wu , Chengyun Jin , Bin Yu . 生物化学在高水平创新型药学人才培养中的交叉融合应用——以去甲基化酶LSD1抑制剂的活性评价为例. University Chemistry, 2025, 40(6): 341-352. doi: 10.12461/PKU.DXHX202406018
17. [17]
  Heng Zhang . Determination of All Rate Constants in the Enzyme Catalyzed Reactions Based on Michaelis-Menten Mechanism. University Chemistry, 2024, 39(4): 395-400. doi: 10.3866/PKU.DXHX202310047
18. [18]
  Chao Liu , Huan Yu , Jiaming Li , Xi Yu , Zhuangzhi Yu , Yuxi Song , Feng Zhang , Qinfang Zhang , Zhigang Zou . 具有光热效应的多级Ti₃C₂/Bi₁₂O₁₇Br₂肖特基异质结简单合成及其太阳能驱动抗生素光降解的研究. Acta Physico-Chimica Sinica, 2025, 41(7): 100075-0. doi: 10.1016/j.actphy.2025.100075
19. [19]
  Yu Dai , Xueting Sun , Haoyu Wu , Naizhu Li , Guoe Cheng , Xiaojin Zhang , Fan Xia . Determination of the Michaelis Constant for Gold Nanozyme-Catalyzed Decomposition of Hydrogen Peroxide. University Chemistry, 2025, 40(5): 351-356. doi: 10.12461/PKU.DXHX202407052
20. [20]
  Chunmei GUO , Weihan YIN , Jingyi SHI , Jianhang ZHAO , Ying CHEN , Quli FAN . Facile construction and peroxidase-like activity of single-atom platinum nanozyme. Chinese Journal of Inorganic Chemistry, 2024, 40(9): 1633-1639. doi: 10.11862/CJIC.20240162

Get Citation

PDF

XML

Metrics

PDF Downloads(1432)
Abstract views(2630)
HTML views(98)

通讯作者: 陈斌, bchen63@163.com

1.
沈阳化工大学材料科学与工程学院沈阳 110142

Address：Zhongguancun North First Street 2,100190 Beijing, PR China Tel: +86-010-82449177-888

DownLoad: Full-Size Img PowerPoint

Return