A novel method for synthesizing authentic SARS-CoV-2 main protease.

Autor: 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.
Jazyk: angličtina
Zdroj: Protein expression and purification [Protein Expr Purif] 2024 Oct; Vol. 222, pp. 106531. Date of Electronic Publication: 2024 Jun 08.
DOI: 10.1016/j.pep.2024.106531
Abstrakt: The SARS-CoV-2 main protease (M pro ) plays a crucial role in virus amplification and is an ideal target for antiviral drugs. Currently, authentic M pro is prepared through two rounds of proteolytic cleavage. In this method, M pro 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 M pro through single digestion. M pro 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 M pro generated by the previous method. These findings indicated that authentic M pro was successfully obtained. Moreover, the substrate specificity of M pro was investigated. M pro 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 M pro for sudden coronavirus infection in the future.
Competing Interests: Declaration of competing interest The authors declare no conflict of interest.
(Copyright © 2024 Elsevier Inc. All rights reserved.)
Databáze: MEDLINE
Externí odkaz: