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A High-Throughput Radioactivity-Based Assay for Screening SARS-CoV-2 nsp10-nsp16 Complex
Aliakbar Khalili Yazdi1, Fengling Li1, Kanchan Devkota1
1Structural Genomics Consortium, University of Toronto, Toronto, Ontario, Canada.
SLAS Discovery : Advancing Life Sciences R & D
|April 20, 2021
Summary
Researchers developed a new assay to screen for drugs targeting the SARS-CoV-2 nsp10-nsp16 complex, crucial for coronavirus replication. This high-throughput screening assay could accelerate the discovery of broad-spectrum antiviral therapies against emerging coronaviruses.
Area of Science:
- Virology
- Drug Discovery
- Biochemistry
Background:
- Novel coronaviruses (CoVs), including SARS-CoV-2, pose significant global health threats requiring rapid therapeutic development.
- Coronaviruses evade host immunity via mRNA capping, a process involving the nsp10-nsp16 complex's 2'-O-methylation activity, making it a key antiviral target.
Purpose of the Study:
- To develop and optimize a high-throughput screening (HTS) assay for the methyltransferase activity of the SARS-CoV-2 nsp10-nsp16 complex.
- To enable efficient screening of potential antiviral drug candidates targeting this conserved viral enzyme.
Main Methods:
- Development of a radioactivity-based assay for nsp10-nsp16 methyltransferase activity.
- Assay optimization and validation in a 384-well format.
- Kinetic characterization of the enzyme-assay system.
Main Results:
- A robust, radioactivity-based HTS assay for SARS-CoV-2 nsp10-nsp16 was successfully developed.
- The assay demonstrated high performance with a Z' factor of 0.83, suitable for HTS.
- Kinetic characterization provided insights into the enzyme's activity.
Conclusions:
- The developed assay addresses the unmet need for efficient screening of SARS-CoV-2 nsp10-nsp16 inhibitors.
- Targeting the conserved nsp16 enzyme may yield broad-spectrum antiviral therapeutics effective against various CoVs.
