Fig. 5
From: Discovery of effectors for casein kinase signaling in the African trypanosome
Substitution of alanine for serine 72 decreases CK1.2 phosphorylation of a BBP59 peptide. (A) Alanine scanning mutants of the BBP59 peptide substrate of CK1.2. (B) Peptides were tested as substrates as sets in three independent experiments. Each peptide was tested in two technical replicates per independent experiment. Symbols represent mean counts per minute of technical replicates and are color and shape-coded according to the experimental replicate: blue triangles, red diamonds, and black squares represent mean counts per minute (CPM) from independent replicate sets. Lines represent mean CPM values. A repeated measures one-way ANOVA with Dunnett’s multiple comparisons test was used to test for significance between each peptide and the control (“Original”) (“Silenced”, p = < 0.0001, “No Substrate,” p = < 0.0001, S66A, p = 3.5 × 10–3; S69A, p = 2.8 × 10–3; T70A, p = 0.0993; S72A, p = < 0.0001). (C) Linear peptide motifs within CK1.2 substrates. Amino acids encompassing substrates of CK1.2 were analyzed for patterns using PRATT. Search parameters were C = 100, PL = 20, PN = 20, PX = 5, FN = 3, FL = 3, E = 3. Two motifs predicting the Ser72 residue of BBP59 are presented along with alignment of substrates for CK1.2 (from Fig. 2). The sequence of ZC3H11 protein was searched using ScanProsite, with CK1.2 target motifs as queries. Scan was run in match mode (greedy, overlaps, no includes).
