Fig. 2: Neuro-phosphoproteomic analysis using non-specific biotinylation enzyme, TurboID.
From: Molecular encoding and synaptic decoding of context during salt chemotaxis in C. elegans
a Schematic illustration of neuro-phosphoproteomic analysis using TurboID. b Biotin label in rgef-1p::TurboID was visualized using streptavidin-Alexa. The label appeared to be restricted in the nervous system. At least two animals observed by confocal microscopy showed similar pattern, with multiple animals appeared to be similarly stained under fluorescence microscopy. Scale bar: 40 µm. c Results of neuro-phosphoproteomic analysis. pkc-1(nj3[W218stop], loss of function) and pan-neuronal H20p::pkc-1(A160E gain of function)-expressing strains were compared. Volcano plot of phosphoproteomic analysis. Red and blue dots represent upregulated and downregulated (P < 0.05 in two-sided t-test) phosphopeptides, respectively, in pkc-1(gf) expressed strains, compared to pkc-1(nj3lf) mutants. Other phosphopeptides are indicated as gray dots. The purple dot represents the peptide KHSAILSNPVNDQK, which is mapped onto UNC-64. The average of the triplicates (n = 3 independent experiments) is described. Peptides which were not detected in either strains were not represented. d The amount of phosphopeptides derived from PKC-1. e The amount of phosphopeptides from a known PKC substrate, DKF-2. f The amount of phosphopeptides containing UNC-64 Ser65. g The amount of total peptides containing UNC-64. Error bars indicate SEM for all panels in Fig. 2. Source data are provided as Supplemental Data.
