An in-solution snapshot of SARS-COV-2 main protease maturation process and inhibition.
An in-solution snapshot of SARS-COV-2 main protease maturation process and inhibition.
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DOI：
10.1038/s41467-023-37035-5
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发表时间：
2023-03-20
影响因子：
16.6
通讯作者：
Godoy, Andre Schutzer
中科院分区：
文献类型：
--
作者：
Noske, Gabriela Dias;Song, Yun;Fernandes, Rafaela Sachetto;Chalk, Rod;Elmassoudi, Haitem;Koekemoer, Lizbe;Owen, C. David J.;El-Baba, Tarick V.;Robinson, Carol;Oliva, Glaucius;Godoy, Andre Schutzer
SARS-CoV-2 (Mpro) 的主要蛋白酶负责病毒多蛋白的切割。 Mpro 的自我加工称为成熟，它对于酶的二聚化和活性至关重要。在这里，我们使用 C145S Mpro 研究溶液中 N 端裂解的结构和动力学。天然质谱分析表明，混合寡聚态由裂解和未裂解的颗粒组成，表明 N 末端加工对于二聚化并不重要。 3.5 Å 冷冻电镜结构提供了晶体环境限制之外 Mpro N 末端裂解的详细信息。我们表明，不同类别的抑制剂改变了寡聚状态之间的平衡。非共价抑制剂 MAT-POS-e194df51-1 可防止二聚化，而共价抑制剂 nirmatrelvir 则可诱导单体转化为二聚体，即使 N 末端完整。我们的数据表明，Mpro 二聚化是由底物加工过程中的共价连接而不是 N 端加工过程中的诱导契合触发的。 SARS-CoV-2 的主要蛋白酶是病毒复制的关键，但其成熟过程仍不完全清楚。在这里，作者不仅揭示了成熟第一步的独特细节，还展示了不同类别的抑制剂如何阻止这一步骤。
The main protease from SARS-CoV-2 (Mpro) is responsible for cleavage of the viral polyprotein. Mpro self-processing is called maturation, and it is crucial for enzyme dimerization and activity. Here we use C145S Mpro to study the structure and dynamics of N-terminal cleavage in solution. Native mass spectroscopy analysis shows that mixed oligomeric states are composed of cleaved and uncleaved particles, indicating that N-terminal processing is not critical for dimerization. A 3.5 Å cryo-EM structure provides details of Mpro N-terminal cleavage outside the constrains of crystal environment. We show that different classes of inhibitors shift the balance between oligomeric states. While non-covalent inhibitor MAT-POS-e194df51-1 prevents dimerization, the covalent inhibitor nirmatrelvir induces the conversion of monomers into dimers, even with intact N-termini. Our data indicates that the Mpro dimerization is triggered by induced fit due to covalent linkage during substrate processing rather than the N-terminal processing. The Main protease of SARS-CoV-2 is key for viral replication, but its maturation process is still not completely understood. Here, the authors not only reveal unique details from the first step of maturation, but also demonstrate how different classes of inhibitor can block this step.
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影响因子：
21.1
作者：
Chen S;Jonas F;Shen C;Hilgenfeld R
通讯作者：
Hilgenfeld R
影响因子：
3.7
作者：
Ho BL;Cheng SC;Shi L;Wang TY;Ho KI;Chou CY
通讯作者：
Chou CY
DOI：
10.1074/jbc.m109.095851
发表时间：
2010-09-03
期刊：
The Journal of biological chemistry
影响因子：
--
作者：
Li C;Qi Y;Teng X;Yang Z;Wei P;Zhang C;Tan L;Zhou L;Liu Y;Lai L
通讯作者：
Lai L
DOI：
10.1074/jbc.ac120.016154
发表时间：
2020-12-11
期刊：
The Journal of biological chemistry
影响因子：
--
作者：
Kneller DW;Phillips G;Weiss KL;Pant S;Zhang Q;O'Neill HM;Coates L;Kovalevsky A
通讯作者：
Kovalevsky A
影响因子：
5.6
作者：
Cho, Eunice;Rosa, Margarida;Haider, Shozeb
通讯作者：
Haider, Shozeb