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A novel method for synthesizing authentic SARS-CoV-2 main protease.
Zhao, Cheng; Rong, Yi; Shi, Shuyuan; Gao, Wen-Chao; Zhang, Chaofeng.
Afiliación
  • Zhao C; College of Biomedical Engineering, Taiyuan University of Technology, Taiyuan, 030024, People's Republic of China. Electronic address: zhaocheng@tyut.edu.cn.
  • Rong Y; College of Biomedical Engineering, Taiyuan University of Technology, Taiyuan, 030024, People's Republic of China.
  • Shi S; College of Biomedical Engineering, Taiyuan University of Technology, Taiyuan, 030024, People's Republic of China.
  • Gao WC; College of Biomedical Engineering, Taiyuan University of Technology, Taiyuan, 030024, People's Republic of China.
  • Zhang C; College of Biomedical Engineering, Taiyuan University of Technology, Taiyuan, 030024, People's Republic of China. Electronic address: zhangchaofeng@tyut.edu.cn.
Protein Expr Purif ; 222: 106531, 2024 Oct.
Article en En | MEDLINE | ID: mdl-38852715
ABSTRACT
The SARS-CoV-2 main protease (Mpro) plays a crucial role in virus amplification and is an ideal target for antiviral drugs. Currently, authentic Mpro is prepared through two rounds of proteolytic cleavage. In this method, Mpro carries a self-cleavage site at the N-terminus and a protease cleavage site followed by an affinity tag at the C-terminus. This article proposes a novel method for producing authentic Mpro through single digestion. Mpro was constructed by fusing a His tag containing TEV protease cleavage sites at the N-terminus. The expressed recombinant protein was digested by TEV protease, and the generated protein had a decreased molecular weight and significantly increased activity, which was consistent with that of authentic Mpro generated by the previous method. These findings indicated that authentic Mpro was successfully obtained. Moreover, the substrate specificity of Mpro was investigated. Mpro had a strong preference for Phe at position the P2, which suggested that the S2 subsite was an outstanding target for designing inhibitors. This article also provides a reference for the preparation of Mpro for sudden coronavirus infection in the future.
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Texto completo: 1 Banco de datos: MEDLINE Asunto principal: Proteasas 3C de Coronavirus / SARS-CoV-2 Límite: Humans Idioma: En Revista: Protein Expr Purif Asunto de la revista: BIOLOGIA MOLECULAR Año: 2024 Tipo del documento: Article

Texto completo: 1 Banco de datos: MEDLINE Asunto principal: Proteasas 3C de Coronavirus / SARS-CoV-2 Límite: Humans Idioma: En Revista: Protein Expr Purif Asunto de la revista: BIOLOGIA MOLECULAR Año: 2024 Tipo del documento: Article