High-Throughput Activity Assay for Screening Inhibitors of the SARS-CoV-2 Mac1 Macrodomain

Morgan Dasovich, Junlin Zhuo, Jack A. Goodman, Ajit Thomas, Robert Lyle McPherson, Aravinth Kumar Jayabalan, Veronica F. Busa, Shang Jung Cheng, Brennan A. Murphy, Karli R. Redinger, Yousef M.O. Alhammad, Anthony R. Fehr, Takashi Tsukamoto, Barbara S. Slusher, Jürgen Bosch, Huijun Wei, Anthony K.L. Leung

Research output: Contribution to journalArticlepeer-review


Macrodomains are a class of conserved ADP-ribosylhydrolases expressed by viruses of pandemic concern, including coronaviruses and alphaviruses. Viral macrodomains are critical for replication and virus-induced pathogenesis; therefore, these enzymes are a promising target for antiviral therapy. However, no potent or selective viral macrodomain inhibitors currently exist, in part due to the lack of a high-throughput assay for this class of enzymes. Here we developed a high-throughput ADP-ribosylhydrolase assay using the SARS-CoV-2 macrodomain Mac1. We performed a pilot screen that identified dasatinib and dihydralazine as ADP-ribosylhydrolase inhibitors. Importantly, dasatinib inhibits SARS-CoV-2 and MERS-CoV Mac1 but not the closest human homologue, MacroD2. Our study demonstrates the feasibility of identifying selective inhibitors based on ADP-ribosylhydrolase activity, paving the way for the screening of large compound libraries to identify improved macrodomain inhibitors and to explore their potential as antiviral therapies for SARS-CoV-2 and future viral threats.

Original languageEnglish (US)
Pages (from-to)17-23
Number of pages7
JournalACS chemical biology
Issue number1
StatePublished - Jan 21 2022

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Medicine


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