Improved SARS-CoV-2 main protease high-throughput screening assay using a 5-carboxyfluorescein substrate.

ABSTRACT: The emergence of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) as a global threat to human health has highlighted the need for the development of novel therapies targeting current and emerging coronaviruses with pandemic potential. The coronavirus main protease (M^pro, also called 3CL^pro) is a validated drug target against coronaviruses and has been heavily studied since the emergence of SARS-CoV-2 in late 2019. Here, we report the biophysical and enzymatic characterization of native M^pro, then characterize the steady-state kinetics of several commonly used FRET substrates, fluorogenic substrates, and six of the 11 reported SARS-CoV-2 polyprotein cleavage sequences. We then assessed the suitability of these substrates for high-throughput scree

SUBMITTER: Legare S 

PROVIDER: S-EPMC8849842 | biostudies-literature | 2022 Apr

REPOSITORIES: biostudies-literature