Quantitative proteomic experiments in recent years became almost routine in many aspects of biology. Particularly the quantification of peptides and corresponding phosphorylated counterparts from a single experiment is highly important for understanding of dynamics of signaling pathways. We developed an analytical method to quantify phosphopeptides (pP) in relation to the quantity of the corresponding non-phosphorylated parent peptides (P). We used mixed-mode solid-phase extraction to purify total peptides from tryptic digest and separated them from most of the phosphorous-containing compounds (e.g., phospholipids, nucleotides) which enhances pP enrichment on TiO2 beads. Phosphoproteomic data derived with this designed method allows quantifying pP/P stoichiometry, and qualifying experimental data for mathematical modeling.
CITATION STYLE
Zakhartsev, M., Pertl-Obermeyer, H., & Schulze, W. X. (2016). From phosphoproteome to modeling of plant signaling pathways. In Methods in Molecular Biology (Vol. 1394, pp. 245–259). Humana Press Inc. https://doi.org/10.1007/978-1-4939-3341-9_18
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