6. Concluding Remarks
Through fusion of innovations in high-throughput chromatographic separations of phosphopeptides, detection by mass spectrometry, and bioinformatic analysis, we have assembled a formidable tool to complement the traditional methodologies typically used to study signaling pathways. Although discovery of a large and dynamic set of cell-derived phosphorylation sites in a single proteomics experiment is an impressive accomplishment, understanding which of these sites participate in a given signaling pathway and the nature of this participation is the fundamental challenge confronting the proteomics researcher. It is simply not feasible to make hundreds of site-directed mutants and the associated knockout mice with current methodologies within a single lab in a reasonable amount of time. Therefore, complimentary high-throughput follow-up strategies must be developed to ascertain whether newly discovered phosphorylation sites participate in pathways and the nature of this participation. Our new approach streamlines the usual signaling pathway analysis paradigm by allowing for the production of mutants of phosphorylation sites and signaling proteins shown to exist within cells as opposed to the usual motif-driven site-directed mutagenesis approach. Also, phosphoproteomic characterization of perturbations of global phosphorylation patterns in mutant cells will likely provide a useful perspective on signaling pathways and ultimately a more rapid understanding of the molecular basis of a wide array of diseases.
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Cao, L., Yu, K., Salomon, A.R. (2006). Phosphoproteomic Analysis of Lymphocyte Signaling. In: Tsoukas, C. (eds) Lymphocyte Signal Transduction. Advances in Experimental Medicine and Biology, vol 584. Springer, Boston, MA. https://doi.org/10.1007/0-387-34132-3_19
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DOI: https://doi.org/10.1007/0-387-34132-3_19
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