Chemical pulldown combined with mass spectrometry to identify the molecular targets of antimalarials in cell-free lysates

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Abstract

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).

Original languageEnglish
Article number102002
Number of pages30
JournalSTAR Protocols
Volume4
Issue number1
Early online date6 Jan 2023
DOIs
Publication statusE-pub ahead of print - 6 Jan 2023

Keywords

  • Cell Biology
  • Cell culture
  • Chemistry
  • Mass Spectrometry
  • Microbiology
  • Molecular/Chemical Probes
  • Protein Biochemistry
  • Proteomics

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