A high-throughput radioactivity-based assay for screening SARS-CoV-2 nsp10-nsp16 complex

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Author Name

Aliakbar Khalili Yazdi

Published 3 Projects

Biochemistry

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Fengling Li

Kanchan Devkota

Published 3 Projects

Biochemistry

Sumera Perveen

Published 3 Projects

Biochemistry

Pegah Ghiabi

Published 3 Projects

Biochemistry

Taraneh Hajian

Published 4 Projects

Biochemistry

Albina Bolotokova

Published 4 Projects

Biochemistry Cell Biology

Masoud Vedadi

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Frequent outbreaks of novel coronaviruses (CoVs), highlighted by the current SARS-CoV-2 pandemic, necessitate the development of therapeutics that could be easily and effectively administered world-wide. The conserved mRNA-capping process enables CoVs to evade their host immune system and is a target for antiviral development. Nonstructural protein (nsp) 16 in complex with nsp10 catalyzes the final step of coronaviral mRNA-capping through its 2'-O-methylation activity. Like other methyltransferases, SARS-CoV-2 nsp10-nsp16 complex is druggable. However, the availability of an optimized assay for high-throughput screening (HTS) is an unmet need. Here, we report the development of a radioactivity-based assay for methyltransferase activity of nsp10-nsp16 complex in a 384-well format, and kinetic characterization, and optimization of the assay for HTS (Z'-factor: 0.83). Considering the high conservation of nsp16 across known CoV species, the potential inhibitors targeting SARS-CoV-2 nsp10-nsp16 complex may also be effective against other emerging pathogenic CoVs.

Biochemistry
Biochemistry 9 Projects