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. 2022 Aug 17;11:e80336. doi: 10.7554/eLife.80336

Figure 6. The PP1-dependent phosphoproteome.

(A) Schematic of the phosphoproteomics time course. PP1-AID parasites were treated with IAA or vehicle for 3 hr. Extracellular parasites were then treated with zaprinast, and samples were collected during the first minute after stimulation. The experiment was performed in biological replicate (R1 and R2). (B) Five clusters were identified with respect to phosphopeptide dynamics and PP1-dependence. (C) Examples of phosphopeptides dynamically regulated by zaprinast and exhibiting PP1-dependent dephosphorylation. (D) GO terms enriched among phosphopeptides, grouped by cluster. Gene ratio is the proportion of proteins with the indicated GO term divided by the total number of proteins belonging to each cluster. Significance was determined with a hypergeometric test; only GO terms with p<0.05 and represented by more than one protein are shown. Redundant GO terms were removed. Cluster 1 lacked enough peptides for enrichment analysis.

Figure 6.

Figure 6—figure supplement 1. Extended analysis of PP1 phosphoproteomics experiment.

Figure 6—figure supplement 1.

(A) Aggregate protein abundances from the non-phosphopeptide enriched samples of parasites treated with IAA or vehicle. Proteins quantified by a single peptide or more are shown in light and dark gray, respectively. Lines correspond to two median absolute deviations. (B) Immunoblot of samples used for the PP1 phosphoproteomics experiment. (C) Quantification of immunoblot band intensity. Intensity was normalized relative to the signal of the vehicle-treated lane for each replicate. Mean ± SD plotted for two independent replicates.
Figure 6—figure supplement 1—source data 1. This file contains the source data that was presented in Figure 6—figure supplement 1.
CDPK1, LICOR 700 channel.
Figure 6—figure supplement 1—source data 2. This file contains the source data that was presented in Figure 6—figure supplement 1.
PP1-HA, LICOR 800 channel.
Figure 6—figure supplement 1—source data 3. This file contains the annotated source data that was presented in Figure 6—figure supplement 1.