A phycobiliprotein-based reporter assay for the evaluation of SARS-CoV-2 main protease activity

IF 2.8 3区医学 Q3 VIROLOGY

Virology Pub Date : 2025-04-12 DOI:10.1016/j.virol.2025.110540

Le Cao , Shuyuan Shi , Chaofeng Zhang , Cheng Zhao

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引用次数: 0

Abstract

SARS-CoV-2 M^pro is crucial for viral replication and transcription and is highly conserved. Therefore, it is an ideal target for developing broad-spectrum antiviral drugs. To address resistance to existing drugs caused by mutations, a simple and sensitive method for detecting the activity of M^pro is needed. Considering the excellent fluorescence properties of phycobiliproteins, this study developed a phycobiliprotein-based reporter assay to evaluate M^pro activity. An engineered lyase was generated by inserting the M^pro recognition sequence between the phycobiliprotein lyases CpcF and CpcE. To ensure that the binding of CpcE and CpcF depended on the linker, a series of truncated forms were constructed. Among them, the activity of CpcE/F-10 was significantly reduced in the presence of M^pro; however, both genetic and chemical inhibition of M^pro activity reversed these results. These data indicated that the fluorescence of phycobiliproteins was negatively correlated with M^pro activity. The reporter assay developed here will contribute to determining the impact of M^pro mutations and screening for new inhibitors.

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来源期刊

Virology 医学-病毒学

CiteScore

6.00

自引率

0.00%

发文量

157

审稿时长

50 days

期刊介绍： Launched in 1955, Virology is a broad and inclusive journal that welcomes submissions on all aspects of virology including plant, animal, microbial and human viruses. The journal publishes basic research as well as pre-clinical and clinical studies of vaccines, anti-viral drugs and their development, anti-viral therapies, and computational studies of virus infections. Any submission that is of broad interest to the community of virologists/vaccinologists and reporting scientifically accurate and valuable research will be considered for publication, including negative findings and multidisciplinary work.Virology is open to reviews, research manuscripts, short communication, registered reports as well as follow-up manuscripts.