Protocol to identify flavonoid antagonists of the SARS-CoV-2 main protease

Moua Yang; Lin Lin; Robert Flaumenhaft

doi:10.1016/j.xpro.2024.102990

Protocol to identify flavonoid antagonists of the SARS-CoV-2 main protease

STAR Protoc. 2024 Apr 6;5(2):102990. doi: 10.1016/j.xpro.2024.102990. Online ahead of print.

Authors

Moua Yang¹, Lin Lin², Robert Flaumenhaft³

Affiliations

¹ Division of Hemostasis and Thrombosis, Beth Israel Deaconess Medical Center and Harvard Medical School, Boston, MA 02115, USA. Electronic address: myang4@bidmc.harvard.edu.
² College of Materials and Chemical Engineering, Minjiang University, Fuzhuo Institute of Oceanography, Fuzhuo, China.
³ Division of Hemostasis and Thrombosis, Beth Israel Deaconess Medical Center and Harvard Medical School, Boston, MA 02115, USA. Electronic address: rflaumen@bidmc.harvard.edu.

Abstract

Flavonoids are naturally occurring metabolites of plants that can inhibit the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) main protease (Mpro), which is required for viral replication. Here, we present a protocol to identify flavonoid antagonists of the SARS-CoV-2 Mpro. We describe steps for the expression and purification of Mpro and a kinetic enzymatic assay for Mpro activity using a dequenching fluorescence resonance energy transfer peptide substrate. We then detail procedures for using this enzymatic assay to test flavonoid antagonism and reversible inhibition. For complete details on the use and execution of this protocol, please refer to Lin et al.¹.

Keywords: High Throughput Screening; Molecular Biology; Protein Biochemistry; Protein expression and purification.

Grants and funding

K99 HL164888/HL/NHLBI NIH HHS/United States