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

Etienne Caron, Romain Roncagalli, Takeshi Hase, Witold E. Wolski, Meena Choi, Marisa G. Menoita, Stephane Durand, Antonio García-Blesa, Ivo Fierro-Monti, Tatjana Sajic, Moritz Heusel, Tobias Weiss, Marie Malissen, Ralph Schlapbach, Ben C. Collins, Samik Ghosh, Hiroaki Kitano, Ruedi Aebersold, Bernard Malissen, Matthias Gstaiger

Research output: Contribution to journalArticlepeer-review

20 Citations (Scopus)
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Abstract

Summary 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.
Original languageEnglish
Pages (from-to)3219-3226
Number of pages8
JournalCell Reports
Volume18
Issue number13
Early online date28 Mar 2017
DOIs
Publication statusEarly online date - 28 Mar 2017

Keywords

  • Targeted mass spectrometry
  • DIA
  • SWATH
  • interactome
  • GRB2
  • primary T cells

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