Robert J Smith , Rachel Milne , Victoriano Corpas Lopez , Natalie Wiedemar , Gourav Dey , Aisha J Syed , Stephen Patterson , Susan Wyllie
STAR Protoc. 2023 Jan 6;4(1):102002. Epub ahead of print. PMID: 36609153.
Here, we provide a protocol using chemical pulldown combined with mass spectrometry (LC-MS/MS) to identify drug targets in Plasmodium falciparum. This approach works upon the principle that a resin-bound inhibitor selectively binds its molecular target(s) in cell-free lysates. We describe the preparation of drug beads and P. falciparum lysate, followed by chemical pulldown, sample fractionation, and LC-MS/MS analysis. We then detail how to identify specifically bound proteins by comparing protein enrichment in DMSO-treated relative to drug-treated lysates via quantitative proteomics. For complete details on the use and execution of this protocol, please refer to Milne et al. (2022).