Figure 1: A schematic representation of the quantitative global phosphoproteomic study to determine the cellular targets for PfPKG.

(a) Late schizont stage parasites were purified on MACS column and incubated with or without Compound 2 (2 μM, 60 min) after which parasite lysates were prepared and digested with trypsin. Peptide samples from individual experiments were labelled with isobaric mass tags and the peptides from three independent experiments were pooled, then phosphopeptides were enriched by sequential fractionation using anion-exchange, PHOS-select (IMAC) and TiO₂ beads before being analysed by LC-MS/MS. (b,c) Illustrative mass spectra showing an example of a phosphopeptide that was less abundant following Compound 2 treatment (b) and one that was unchanged by Compound 2 treatment (c).