Figure 2
Impact of STS on the non-kinome and phosphoproteome. (A) The number of hits identified as kinases, and/or localized to the mitochondria, related to Wnt signaling and/or apoptosis. (B) Changes in PWP1 protein complex upon STS treatment as analysed by MS-immunoprecipitation. Hit proteins are defined by average log2 fold change (logFC) < − 0.5 or > 0.5, adjusted p-value < 0.05, from three replicates. PSM (number of peptide-spectra match per protein). CAMKK2 and AMPK were identified in the ProteomeImpact experiment (Fig. 1F), with CAMKK2 identified as a direct interactor of STS (solid purple fill). Known regulation of AMPK by CAMKK227,28,29,30 (as indicated by the faint purple fill and dotted lines). (C) Log 2 abundance intensity of the total protein (ACACA) and its phosphosite ACACA-Ser80 with (STS) and without (CTRL) treatment. (D) Phosphopeptide response curves for four proteins with all three response patterns: thyroid hormone receptor-associated protein 3 (THRAP3), DNA topoisomerase 2-beta (TOP2B), RalBP1-associated Eps domain-containing protein 1 (REPS1), small glutamine-rich tetratricopeptide repeat-containing protein alpha (SGTA). The doses were represented as ordinal numbers, with the highest dose 10.0 µM at position number 10 (x-axis). The ptm (post-translation modification) abundance 37C (y-axis) represents the abundance fold change relative to the untreated condition. Coloured bars indicate biphasic (yellow), hyper-phosphorylation (red) and hypo-phosphorylation (blue), green and brown lines represent two biological replicates. (E) Differential phosphosites are clustered by biological processes involved. DE size (differential expression size), significance (enrichment significance). (F) Signaling network of epidermal growth factor receptor (EGFR) and insulin receptor (IR). Proteins with biphasic phosphorylation response in this pathway (shaded yellow).
