title: Characterization of Receptor-Associated Protein Complex Assembly in Interleukin (IL)-2- and IL-15-Activated T-Cell Lines.
abstract: It remains a paradox that IL-2 and IL-15 can differentially modulate the immune response using the same signaling receptors. We have previously dissected the phosphotyrosine-driven signaling cascades triggered by both cytokines in Kit225 T-cells, unveiling subtle differences that may contribute to their functional dichotomy. In this study, we aimed to decipher the receptor complex assembly in IL-2- and IL-15-activated T-lymphocytes that is highly orchestrated by site-specific phosphorylation events. Comparing the cytokine-induced interactome of the interleukin receptor beta and gamma subunits shared by the two cytokines, we defined the components of the early IL-2 and IL-15 receptor-associated complex discovering novel constituents. Additionally, phosphopeptide-directed analysis allowed us to detect several cytokine-dependent and -independent phosphorylation events within the activated receptor complex including novel phosphorylated sites located in the cytoplasmic region of IL-2 receptor β subunit (IL-2Rβ). We proved that the distinct phosphorylations induced by the cytokines serve for recruiting different types of effectors to the initial receptor/ligand complex. Overall, our study sheds new light into the initial molecular events triggered by IL-2 and IL-15 and constitutes a further step toward a better understanding of the early signaling aspects of the two closely related cytokines in T-lymphocytes.
authors.affiliation: Department of Biochemistry and Molecular Biology, University of Southern Denmark , 5230 Odense M, Denmark.
authors.forename: Nerea
authors.initials: N
authors.lastname: Osinalde
authors.affiliation: Department of Biochemistry and Molecular Biology, University of Southern Denmark , 5230 Odense M, Denmark.
authors.forename: Virginia
authors.initials: V
authors.lastname: Sanchez-Quiles
authors.affiliation: Department of Biochemistry and Molecular Biology, University of Southern Denmark , 5230 Odense M, Denmark.
authors.forename: Vyacheslav
authors.initials: V
authors.lastname: Akimov
authors.affiliation: Proteomics Core Facility-SGIKER, University of the Basque Country, UPV/EHU , 48940 Leioa, Spain.
authors.forename: Kerman
authors.initials: K
authors.lastname: Aloria
authors.affiliation: Department of Biochemistry and Molecular Biology, University of the Basque Country, UPV/EHU , 48940 Leioa, Spain.
authors.forename: Jesus M
authors.initials: JM
authors.lastname: Arizmendi
authors.affiliation: Department of Biochemistry and Molecular Biology, University of Southern Denmark , 5230 Odense M, Denmark.
authors.forename: Blagoy
authors.initials: B
authors.lastname: Blagoev
authors.affiliation: Department of Biochemistry and Molecular Biology, University of Southern Denmark , 5230 Odense M, Denmark.
authors.forename: Irina
authors.initials: I
authors.lastname: Kratchmarova
mesh_terms: D000595:Amino Acid Sequence; D045744:Cell Line, Tumor; D005786:Gene Expression Regulation; D006801:Humans; D053631:Interleukin Receptor Common gamma Subunit / Q000235:genetics / Q000276:immunology*; D019409:Interleukin-15 / Q000235:genetics / Q000276:immunology / Q000494:pharmacology*; D007376:Interleukin-2 / Q000235:genetics / Q000276:immunology / Q000494:pharmacology*; D053646:Interleukin-2 Receptor beta Subunit / Q000235:genetics / Q000276:immunology*; D053613:Janus Kinase 1 / Q000235:genetics / Q000276:immunology*; D053616:Janus Kinase 3 / Q000235:genetics / Q000276:immunology*; D008213:Lymphocyte Activation; D010766:Phosphorylation; D019000:Phosphotyrosine / Q000235:genetics / Q000276:immunology; D011485:Protein Binding; D025941:Protein Interaction Mapping; D011994:Recombinant Proteins / Q000235:genetics / Q000276:immunology / Q000494:pharmacology; D015398:Signal Transduction; D013601:T-Lymphocytes / Q000166:cytology / Q000187:drug effects* / Q000276:immunology
chemical_list: C569274:IL15 protein, human; C508020:IL2RB protein, human; C507941:IL2RG protein, human; D053631:Interleukin Receptor Common gamma Subunit; D019409:Interleukin-15; D007376:Interleukin-2; D053646:Interleukin-2 Receptor beta Subunit; D011994:Recombinant Proteins; D019000:Phosphotyrosine; C507904:JAK1 protein, human; C507933:JAK3 protein, human; D053613:Janus Kinase 1; D053616:Janus Kinase 3
keywords: SILAC; T-lymphocyte; affinity enrichment; cell signaling; interleukin; phosphoproteomics; protein complexes; quantitative proteomics