Journal: PLoS Neglected Tropical Diseases
Article Title: Deep phosphoproteome analysis of Schistosoma mansoni leads development of a kinomic array that highlights sex-biased differences in adult worm protein phosphorylation
doi: 10.1371/journal.pntd.0008115
Figure Lengend Snippet: Kinomic peptide array ‘chips’ comprising unique substrate peptides (‘spots’) for S . mansoni protein kinases were custom developed on CelluSpots slides and used to screen adult male and female protein kinase activities as detailed in Methods. (A) Representative peptide array slide incubated with female worm homogenate; 93 spots, each containing a unique peptide, were phosphorylated by worm kinases and detected using Phos-tag biotin/streptavidin HRP and ECL. Each slide comprises two sub-arrays each with identical peptides (for layout see ); the graph illustrates regression analysis of the technical replicates (normalized spot intensities) on the array. (B) Comparison of peptide sub-arrays incubated with equivalent amounts of protein (100 μg) from adult male or female worms to illustrate the different kinomic profiles of the sexes. (C) Phosphorylation of a substrate peptide sequence from the protein AMPKL/CAMK by adult male or female worm extracts; the spots shown for each sex (location highlighted in (B), red box) are from four sub-arrays; the phosphorylateable Thr residue is in red. (D) Spot intensities representing phosphorylation of each unique substrate peptide were determined using the ImageJ plugin “Protein Array Analyzer” and normalized; mean relative phosphorylation values (n = 4 ± S.E.M.) are shown and differences between mean male and female kinase activities were evaluated using Student’s t-test (green P<0.05; purple P<0.10). For each substrate peptide the putative upstream kinase (where known) is annotated within brackets. The peptide substrate in (C) (Tyrosine kinase Src) is highlighted (red box).
Article Snippet: The final S . mansoni custom “CelluSpots” 96-spot peptide array was printed under contract by Intavis Bioanalytical Instruments AG.
Techniques: Peptide Microarray, Incubation, Comparison, Phospho-proteomics, Sequencing, Residue, Protein Array