Quantitative phosphoproteomics in cell signaling

Blagoy Blagoev (University of Southern Denmark)

Abstract

The lecture will focus on mass spectrometry-based quantitative proteomics approaches used to characterize and compare signaling networks initiated by receptor tyrosine kinases (RTKs) with emphasis on dynamic phosphorylation(1). The signaling by RTKs is a sequence of reverse phosphorylation events with an initial flow of tyrosine phosphorylation which triggers consequent downstream cascades of mainly threonine and serine phosphorylations that may have positive as well as negative effects on the signaling transduction. The cellular outcome of any given network is depended entirely on the series of induced phosphorylation, thus a creation of global phosphorylation profiles is an essential step towards understanding the mechanisms of regulation of the signaling cascades. Quantitative comparison of entire signaling pathways initiated by different signals will be presented as well as the very early phosphorylation changes that occur in cells within seconds of treatment with ligand(2,3). In addition, the temporal dynamics of phosphorylation-dependent signaling networks using SILAC-based quantitative proteomic approach will be described(4,5). The creation of temporal maps allows addition of another layer of understanding of the complex and dynamic signaling networks induced by growth factors and permits modeling of the signaling pathways in the view of systems biology.

References

1. Blagoev, B. & Mann, M. Quantitative proteomics to study mitogen-activated protein kinases. Methods 40, 243-50 (2006).
2. Dengjel, J. et al. Quantitative proteomic assessment of very early cellular signaling events. Nat Biotechnol 25, 566-8 (2007).
3. Kratchmarova, I. et al. Mechanism of divergent growth factor effects in mesenchymal stem cell differentiation. Science 308, 1472-7 (2005).
4. Blagoev, B. et al. Temporal analysis of phosphotyrosine-dependent signaling networks by quantitative proteomics. Nat Biotechnol 22, 1139-45 (2004).
5. Olsen, J. V. et al. Global, in vivo, and site-specific phosphorylation dynamics in signaling networks. Cell 127, 635-48 (2006).

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