Allosteric Inhibition of the SARS-CoV-2 Main Protease: Insights from Mass Spectrometry Based Assays
Allosteric Inhibition of the SARS-CoV-2 Main Protease: Insights from Mass Spectrometry Based Assays
复制标题
DOI：
10.1002/anie.202010316
复制
发表时间：
2020-10-15
影响因子：
16.6
通讯作者：
Robinson, Carol V.
中科院分区：
文献类型：
--
作者：
El-Baba, Tarick J.;Lutomski, Corinne A.;Robinson, Carol V.
SARS-CoV-2主要蛋白酶(M-Pro)沿两种病毒多肽裂解，释放病毒复制所需的非结构蛋白。M-Pro是抗病毒治疗的一个有吸引力的目标，以对抗冠状病毒-2019年疾病。在这里，我们使用天然质谱学来表征M-PRO的功能单位。单体/二聚体平衡分析表明，M-Pro的离解常数为K-d=0.14+/-0.03µM，表明M-Pro具有较强的溶液二聚倾向。我们通过跟踪酶-底物复合体中的时间变化来表征底物周转率，并筛选出远离活性部位结合的小分子，以了解它们调节活性的能力。这些化合物，包括一种被认为破坏二聚体的化合物，使底物处理速度减慢约35%。这些信息，加上对X射线晶体结构的分析，为开发更有效的分子提供了一个起点，这些分子可以通过变构来调节M-Pro的活性。
The SARS-CoV-2 main protease (M-pro) cleaves along the two viral polypeptides to release non-structural proteins required for viral replication. M-Pro is an attractive target for antiviral therapies to combat the coronavirus-2019 disease. Here, we used native mass spectrometry to characterize the functional unit of M-pro. Analysis of the monomer/dimer equilibria reveals a dissociation constant of K-d=0.14 +/- 0.03 mu M, indicating M-Pro has a strong preference to dimerize in solution. We characterized substrate turnover rates by following temporal changes in the enzyme-substrate complexes, and screened small molecules, that bind distant from the active site, for their ability to modulate activity. These compounds, including one proposed to disrupt the dimer, slow the rate of substrate processing by approximate to 35 %. This information, together with analysis of thex-ray crystal structures, provides a starting point for the development of more potent molecules that allosterically regulate M-Pro activity.