Precise Temporal Profiling of Signaling Complexes in Primary Cells Using SWATH Mass Spectrometry - CNRS - Centre national de la recherche scientifique Accéder directement au contenu
Article Dans Une Revue Cell Reports Année : 2017

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

Etienne Caron
  • Fonction : Auteur
Takeshi Hase
  • Fonction : Auteur
Witold E. Wolski
  • Fonction : Auteur
Meena Choi
  • Fonction : Auteur
Marisa G. Menoita
  • Fonction : Auteur
Stéphane Durand
  • Fonction : Auteur
  • PersonId : 759919
  • IdRef : 191995320
Ivo Fierro-Monti
  • Fonction : Auteur
Tatjana Sajic
  • Fonction : Auteur
Moritz Heusel
  • Fonction : Auteur
Tobias Weiss
  • Fonction : Auteur
Marie Malissen
Ben C. Collins
  • Fonction : Auteur
Samik Ghosh
  • Fonction : Auteur
Hiroaki Kitano
  • Fonction : Auteur
Matthias Gstaiger
  • Fonction : Auteur

Résumé

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.

Domaines

Immunologie

Dates et versions

hal-01996176 , version 1 (28-01-2019)

Identifiants

Citer

Etienne Caron, Romain Roncagalli, Takeshi Hase, Witold E. Wolski, Meena Choi, et al.. Precise Temporal Profiling of Signaling Complexes in Primary Cells Using SWATH Mass Spectrometry. Cell Reports, 2017, 18 (13), pp.3219-3226. ⟨10.1016/j.celrep.2017.03.019⟩. ⟨hal-01996176⟩
54 Consultations
0 Téléchargements

Altmetric

Partager

Gmail Facebook X LinkedIn More