A Yeast-Based Drug Discovery Platform To Identify Plasmodium falciparum Type II NADH Dehydrogenase Inhibitors.
Yu CaoChen SunHan WenMengfei WangPan ZhuMengjiao ZhongJian LiXuewei ChenYefeng TangJingwen WangBing ZhouPublished in: Antimicrobial agents and chemotherapy (2021)
Conventional methods utilizing in vitro protein activity assay or in vivo parasite survival to screen for malaria inhibitors suffer from high experimental background and/or inconvenience. Here, we introduce a yeast-based system to facilitate chemical screening for specific protein or pathway inhibitors. The platform comprises several isogeneic Pichia strains that differ only in the target of interest, so that a compound which inhibits one strain but not the other is implicated in working specifically against the target. We used Plasmodium falciparum NDH2 (PfNDH2), a type II NADH dehydrogenase, as a proof of principle to show how well this works. Three isogenic Pichia strains harboring, respectively, exogeneously introduced PfNDH2, its own complex I (a type I NADH dehydrogenase), and PfNDH2 with its own complex I, were constructed. In a pilot screen of more than 2,000 compounds, we identified a highly specific inhibitor that acts on PfNDH2. This compound poorly inhibits the parasites at the asexual blood stage; however, is highly effective in repressing oocyst maturation in the mosquito stage. Our results demonstrate that the yeast cell-based screen platform is feasible, efficient, economical, and has very low background noise. Similar strategies could be extended to the functional screen for interacting molecules of other targets.
Keyphrases
- plasmodium falciparum
- high throughput
- single cell
- drug discovery
- saccharomyces cerevisiae
- escherichia coli
- protein protein
- binding protein
- wastewater treatment
- air pollution
- clinical trial
- recombinant human
- amino acid
- cell wall
- bone marrow
- study protocol
- protein kinase
- cell therapy
- aedes aegypti
- free survival
- small molecule