Validation of solvent proteome profiling for antimalarial drug target deconvolution

Area of Science:

• Proteomics
• Drug Discovery
• Malaria Research

Background:

• Malaria poses a global health challenge, exacerbated by increasing drug resistance.
• Identifying drug targets is crucial for developing new antimalarial therapies.
• Understanding drug mechanisms of action is key for optimizing treatments.

Purpose of the Study:

• To validate solvent-induced proteome profiling (SPP) for identifying drug-protein interactions in Plasmodium falciparum.
• To demonstrate SPP's utility with various antimalarial compounds.
• To introduce novel SPP applications for enhanced target elucidation.

Main Methods:

• Generated solvent denaturation curves for the P. falciparum proteome.
• Applied SPP to analyze five antimalarial compounds (pyrimethamine, atovaquone, cipargamin, MMV1557817, OSM-S-106).
• Developed and tested live-cell treatment SPP and one-pot mixed-drug SPP.

Main Results:

• Successfully identified the validated target of cipargamin as non-SERCA-type Ca2+-transporting P-ATPase (PfATP4).
• Demonstrated live-cell SPP's specificity in identifying pyrimethamine's target, dihydrofolate reductase-thymidylate synthase (PfDHFR).
• Showcased the efficiency of one-pot mixed-drug SPP for evaluating multiple drugs.

Conclusions:

• SPP is a versatile proteomics approach for elucidating antimalarial drug targets in P. falciparum.
• SPP is effective under both lysate and live-cell conditions.
• This method provides insights for optimizing antimalarials and developing novel therapies.