A High-Throughput Radioactivity-Based Assay for Screening SARS-CoV-2 Nsp10-nsp16 Complex

Overview

Journal SLAS Discov

Publisher Sage Publications

Specialty Molecular Biology

Date 2021 Apr 20

PMID 33874769

Citations 18

Authors

Aliakbar Khalili Yazdi

Fengling Li

Kanchan Devkota

Sumera Perveen

Pegah Ghiabi

Taraneh Hajian

Albina Bolotokova

Masoud Vedadi

Affiliations

Soon will be listed here.

Abstract

Frequent outbreaks of novel coronaviruses (CoVs), highlighted by the current severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) pandemic, necessitate the development of therapeutics that could be easily and effectively administered worldwide. 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'--methylation activity. Like other methyltransferases, the 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 the methyltransferase activity of the nsp10-nsp16 complex in a 384-well format, 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 the SARS-CoV-2 nsp10-nsp16 complex may also be effective against other emerging pathogenic CoVs.

Citing Articles

Cannabinoid-Inspired Inhibitors of the SARS-CoV-2 Coronavirus 2'--Methyltransferase (2'--MTase) Non-Structural Protein (Nsp10-16).

Benjamin M, Hanna G, Dickinson C, Choo Y, Wang X, Downs-Bowen J Molecules. 2024; 29(21).

PMID: 39519722 PMC: 11547505. DOI: 10.3390/molecules29215081.

Kinetic characterization of human mRNA guanine-N7 methyltransferase.

Perveen S, Yazdi A, Hajian T, Li F, Vedadi M Sci Rep. 2024; 14(1):4509.

PMID: 38402266 PMC: 10894281. DOI: 10.1038/s41598-024-55184-5.

Informatics and Computational Approaches for the Discovery and Optimization of Natural Product-Inspired Inhibitors of the SARS-CoV-2 2'--Methyltransferase.

Hanna G, Benjamin M, Choo Y, De R, Schinazi R, Nielson S J Nat Prod. 2024; 87(2):217-227.

PMID: 38242544 PMC: 10898454. DOI: 10.1021/acs.jnatprod.3c00875.

Discovery of a Druggable, Cryptic Pocket in SARS-CoV-2 nsp16 Using Allosteric Inhibitors.

Inniss N, Kozic J, Li F, Rosas-Lemus M, Minasov G, Rybacek J ACS Infect Dis. 2023; 9(10):1918-1931.

PMID: 37728236 PMC: 10961098. DOI: 10.1021/acsinfecdis.3c00203.

Coronavirus 2'-O-methyltransferase: A promising therapeutic target.

Schindewolf C, Menachery V Virus Res. 2023; 336:199211.

PMID: 37634741 PMC: 10485632. DOI: 10.1016/j.virusres.2023.199211.

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