Screening of Leishmania braziliensis dihydroorotate dehydrogenase inhibitors

Thermoshift assay is a versatile method to screen compounds. Besides sypro-orange (ThermoFlour), the intrinsic fluorescence of the FMN was also exploited to develop sensitive TSA (ThermoFMN). Aiming at investigating the correlation between those strategies for screening, 400 compounds were screened against LbDHODH using both strategies. Using a Tm  1 C, only 3% of the compounds were found to induce thermoshift in both assays, 26.4% only in ThermoFluor, and 1.9% only in ThermoFMN. The crystallographic structure of LbDHODH in complex with compounds of each thermoshift profile was solved and allowed us to identify ubiquitous electron density for the ligands at different binding pockets.