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Early downstream responses of T lymphocytes following T cell antigen receptor (TCR) activation are mediated by protein complexes that assemble in domains of the plasma membrane. Using stable isotope labeling with amino acids in cell culture and mass spectrometry, we quantitatively related the proteome of αCD3 immunoisolated native TCR signaling plasma membrane domains to that of control plasma membrane fragments not engaged in TCR signaling. Proteins were sorted according to their relative enrichment in isolated TCR signaling plasma membrane domains, identifying a complex protein network that is anchored in the vicinity of activated TCR. These networks harbor widespread mediators of plasma membrane-proximal T cell activities, including propagation, balancing, and attenuation of TCR signaling, immune synapse formation, as well as cytoskeletal arrangements relative to TCR activation clusters. These results highlight the unique potential of systematic characterizations of plasma membrane-proximal T cell activation proteome in the context of its native lipid bilayer platform.

Original publication

DOI

10.1074/jbc.M110.165415

Type

Journal article

Journal

J Biol Chem

Publication Date

11/02/2011

Volume

286

Pages

4072 - 4080

Keywords

Humans, Jurkat Cells, Lymphocyte Activation, Membrane Proteins, Proteome, Receptors, Antigen, T-Cell, Signal Transduction, T-Lymphocytes