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A High-Throughput Radioactivity-Based Assay for Screening SARS-CoV-2 nsp10-nsp16 Complex.
Khalili Yazdi, Aliakbar; Li, Fengling; Devkota, Kanchan; Perveen, Sumera; Ghiabi, Pegah; Hajian, Taraneh; Bolotokova, Albina; Vedadi, Masoud.
  • Khalili Yazdi A; Structural Genomics Consortium, University of Toronto, Toronto, Ontario, Canada.
  • Li F; Structural Genomics Consortium, University of Toronto, Toronto, Ontario, Canada.
  • Devkota K; Structural Genomics Consortium, University of Toronto, Toronto, Ontario, Canada.
  • Perveen S; Structural Genomics Consortium, University of Toronto, Toronto, Ontario, Canada.
  • Ghiabi P; Structural Genomics Consortium, University of Toronto, Toronto, Ontario, Canada.
  • Hajian T; Structural Genomics Consortium, University of Toronto, Toronto, Ontario, Canada.
  • Bolotokova A; Structural Genomics Consortium, University of Toronto, Toronto, Ontario, Canada.
  • Vedadi M; Structural Genomics Consortium, University of Toronto, Toronto, Ontario, Canada.
SLAS Discov ; 26(6): 757-765, 2021 07.
Article in English | MEDLINE | ID: covidwho-1194439
Preprint
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Semantic information from SemMedBD (by NLM)
1. Assay METHOD_OF High-Throughput Screening
Subject
Assay
Predicate
METHOD_OF
Object
High-Throughput Screening
2. Assay METHOD_OF High-Throughput Screening
Subject
Assay
Predicate
METHOD_OF
Object
High-Throughput Screening
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'-O-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.
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Full text: Available Collection: International databases Database: MEDLINE Main subject: RNA Caps / RNA, Viral / Adenosine / Viral Nonstructural Proteins / Viral Regulatory and Accessory Proteins / High-Throughput Screening Assays Type of study: Diagnostic study / Prognostic study / Screening study Language: English Journal: SLAS Discov Year: 2021 Document Type: Article Affiliation country: 24725552211008863

Full text: Available Collection: International databases Database: MEDLINE Main subject: RNA Caps / RNA, Viral / Adenosine / Viral Nonstructural Proteins / Viral Regulatory and Accessory Proteins / High-Throughput Screening Assays Type of study: Diagnostic study / Prognostic study / Screening study Language: English Journal: SLAS Discov Year: 2021 Document Type: Article Affiliation country: 24725552211008863