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Publication : Precise Temporal Profiling of Signaling Complexes in Primary Cells Using SWATH Mass Spectrometry.

First Author  Caron E Year  2017
Journal  Cell Rep Volume  18
Issue  13 Pages  3219-3226
PubMed ID  28355572 Mgi Jnum  J:254051
Mgi Id  MGI:6103695 Doi  10.1016/j.celrep.2017.03.019
Citation  Caron E, et al. (2017) Precise Temporal Profiling of Signaling Complexes in Primary Cells Using SWATH Mass Spectrometry. Cell Rep 18(13):3219-3226
abstractText  Spatiotemporal organization of protein interactions in cell signaling is a fundamental process that drives cellular functions. Given differential protein expression across tissues and developmental stages, the architecture and dynamics of signaling interaction proteomes is, likely, highly context dependent. However, current interaction information has been almost exclusively obtained from transformed cells. In this study, we applied an advanced and robust workflow combining mouse genetics and affinity purification (AP)-SWATH mass spectrometry to profile the dynamics of 53 high-confidence protein interactions in primary T cells, using the scaffold protein GRB2 as a model. The workflow also provided a sufficient level of robustness to pinpoint differential interaction dynamics between two similar, but functionally distinct, primary T cell populations. Altogether, we demonstrated that precise and reproducible quantitative measurements of protein interaction dynamics can be achieved in primary cells isolated from mammalian tissues, allowing resolution of the tissue-specific context of cell-signaling events.
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