Identification of novel Trypanosoma cruzi proteasome inhibitors using a luminescence-based high-throughput screening assay.

Identification of novel Trypanosoma cruzi proteasome inhibitors using a luminescence-based high-throughput screening assay. Antimicrob Agents Chemother. 2019 Jul 15;: Authors: Zmuda F, Sastry L, Shepherd SM, Jones D, Scott A, Craggs PD, Cortes A, Gray DW, Torrie LS, De Rycker M Abstract Chagas' disease, caused by the protozoan parasite Trypanosoma cruzi, is a potentially life-threatening condition that has become a global issue. Current treatment is limited to two medicines that require prolonged dosing and are associated with multiple side effects, which often lead to treatment discontinuation and failure. One way to address these shortcomings is through target-based drug discovery on validated T. cruzi protein targets. One such target is the proteasome, which plays a crucial role in protein degradation and turnover, through chymotrypsin-, trypsin-, and caspase-like catalytic activities. In order to initiate a proteasome drug discovery programme, we isolated proteasomes from T. cruzi epimastigotes and characterized their activity using a commercially available glow-like luminescence-based assay. We developed a high-throughput biochemical assay for the chymotrypsin-like activity of the T. cruzi proteasome, which was found to be sensitive, specific, and robust, but prone to luminescence technology interference. To mitigate this, we have also developed a counter-screen assay that identifies potential interferers at the level of both th...
Source: Antimicrobial Agents and Chemotherapy - Category: Microbiology Authors: Tags: Antimicrob Agents Chemother Source Type: research