). This prompted us to design novel substrates based on the 2A/2B and 2B/3 cleavage sites. Enzyme kinetics and inhibition of the DENV1-4 proteases were characterized under two widely applicable assay conditions. The feasibility to use these conditions for HTS was verified in 384-well format. Based on the results, we present two robust assay protocols, suitable to examine activity and inhibition of proteases of all four DENV serotypes in parallel, allowing for HTS. A well-characterized assay procedure can enhance coherence in DENV protease studies, contributing to progress in antiviral research.
Macedo et al. (Tue,) studied this question.