An isothermal calorimetry assay for determining steady state kinetic and Ensitrelvir inhibition parameters for SARS-CoV-2 3CL-protease.

ABSTRACT: This manuscript details the application of Isothermal Titration Calorimetry (ITC) to characterize the kinetics of 3CL^pro, the main protease from the Severe Acute Respiratory Syndrome CoronaVirus-2 (SARS-CoV-2), and its inhibition by Ensitrelvir, a known non-covalent inhibitor. 3CL^pro is essential for producing the proteins necessary for viral infection, which led to the COVID-19 pandemic. The ITC-based assay provided rapid and reliable measurements of 3CL^pro activity, allowing for the direct derivation of the kinetic enzymatic constants K_M and k_cat by monitoring the thermal power required to maintain a constant temperature as the substrate is consumed. The manuscript highlights several advantages of the proposed ITC-based assay over tr

SUBMITTER: Mazzei L 

PROVIDER: S-EPMC11688438 | biostudies-literature | 2024 Dec

REPOSITORIES: biostudies-literature