The Chinese Academy of Sciences Key Laboratory of Receptor Research, State Key Laboratory of Drug Research, Shanghai Institute of Materia Medica, Chinese Academy of Sciences, Shanghai 201203, China.
Department of Biopharmaceutics, College of Food Science and Technology, Shanghai Ocean University, Shanghai 201306, China.
Proc Natl Acad Sci U S A. 2022 Apr 12;119(15):e2120913119. doi: 10.1073/pnas.2120913119. Epub 2022 Mar 24.
SignificanceThe coronavirus main protease (M) is required for viral replication. Here, we obtained the extended conformation of the native monomer of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) M by trapping it with nanobodies and found that the catalytic domain and the helix domain dissociate, revealing allosteric targets. Another monomeric state is termed compact conformation and is similar to one protomer of the dimeric form. We designed a Nanoluc Binary Techonology (NanoBiT)-based high-throughput allosteric inhibitor assay based on structural conformational change. Our results provide insight into the maturation, dimerization, and catalysis of the coronavirus M and pave a way to develop an anticoronaviral drug through targeting the maturation process to inhibit the autocleavage of M.
意义冠状病毒主蛋白酶 (M) 是病毒复制所必需的。在这里，我们通过纳米体捕获获得了严重急性呼吸系统综合征冠状病毒 2 (SARS-CoV-2) M 的天然单体的扩展构象，并发现催化结构域和螺旋结构域解离，揭示了变构靶标。另一种单体状态称为紧凑构象，类似于二聚体形式的一个单体。我们基于结构构象变化设计了基于 Nanoluc Binary Techonology (NanoBiT) 的高通量变构抑制剂测定法。我们的结果深入了解了冠状病毒 M 的成熟、二聚化和催化过程，并为通过靶向成熟过程来开发抑制 M 自身切割的抗冠状病毒药物铺平了道路。
