title: Interleukin-2 signaling pathway analysis by quantitative phosphoproteomics.
abstract: Interleukin-2 (IL-2) is major cytokine involved in T cell proliferation, differentiation and apoptosis. Association between IL-2 and its receptor (IL-2R), triggers activation of complex signaling cascade governed by tyrosine phosphorylation that culminates in transcription of genes involved in modulation of the immune response. The complete characterization of the IL-2 pathway is essential to understand how aberrant IL-2 signaling results in several diseases such as cancer or autoimmunity and also how IL-2 treatments affect cancer patients. To gain insights into the downstream machinery activated by IL-2, we aimed to define the global tyrosine-phosphoproteome of IL-2 pathway in human T cell line Kit225 using high resolution mass spectrometry combined with phosphotyrosine immunoprecipitation and SILAC. The molecular snapshot at 5min of IL-2 stimulation resulted in identification of 172 proteins among which 79 were found with increased abundance in the tyrosine-phosphorylated complexes, including several previously not reported IL-2 downstream effectors. Combinatorial site-specific phosphoproteomic analysis resulted in identification of 99 phosphorylated sites mapping to the identified proteins with increased abundance in the tyrosine-phosphorylated complexes, of which 34 were not previously described. In addition, chemical inhibition of the identified IL-2-mediated JAK, PI3K and MAPK signaling pathways, resulted in distinct alteration on the IL-2 dependent proliferation.
authors: Department of Biochemistry and Molecular Biology, University of the Basque Country, UPV/EHU, 48940 Leioa, Spain.
authors: Nerea
authors: N
authors: Osinalde
authors: Helle
authors: H
authors: Moss
authors: Onetsine
authors: O
authors: Arrizabalaga
authors: Miren J
authors: MJ
authors: Omaetxebarria
authors: Blagoy
authors: B
authors: Blagoev
authors: Ana M
authors: AM
authors: Zubiaga
authors: Asier
authors: A
authors: Fullaondo
authors: Jesus M
authors: JM
authors: Arizmendi
authors: Irina
authors: I
authors: Kratchmarova
mesh_terms: D045744:Cell Line, Tumor; D006801:Humans; D007376:Interleukin-2 / Q000032:analysis / Q000235:genetics / Q000378:metabolism*; D047908:Intracellular Signaling Peptides and Proteins / Q000032:analysis* / Q000235:genetics / Q000378:metabolism; D015451:Leukemia, Lymphocytic, Chronic, B-Cell / Q000276:immunology / Q000378:metabolism / Q000473:pathology; D010750:Phosphoproteins / Q000032:analysis* / Q000235:genetics / Q000378:metabolism; D040901:Proteomics / Q000379:methods*; D013601:T-Lymphocytes / Q000276:immunology / Q000378:metabolism / Q000473:pathology*
chemical_list: D007376:Interleukin-2; D047908:Intracellular Signaling Peptides and Proteins; D010750:Phosphoproteins