title: Integrative Phosphoproteomics Links IL-23R Signaling with Metabolic Adaptation in Lymphocytes.
abstract: Interleukin (IL)-23 mediated signal transduction represents a major molecular mechanism underlying the pathology of inflammatory bowel disease, Crohn's disease and ulcerative colitis. In addition, emerging evidence supports the role of IL-23-driven Th17 cells in inflammation. Components of the IL-23 signaling pathway, such as IL-23R, JAK2 and STAT3, have been characterized, but elements unique to this network as compared to other interleukins have not been readily explored. In this study, we have undertaken an integrative phosphoproteomics approach to better characterise downstream signaling events. To this end, we performed and compared phosphopeptide and phosphoprotein enrichment methodologies after activation of T lymphocytes by IL-23. We demonstrate the complementary nature of the two phosphoenrichment approaches by maximizing the capture of phosphorylation events. A total of 8202 unique phosphopeptides, and 4317 unique proteins were identified, amongst which STAT3, PKM2, CDK6 and LASP-1 showed induction of specific phosphorylation not readily observed after IL-2 stimulation. Interestingly, quantitative analysis revealed predominant phosphorylation of pre-existing STAT3 nuclear subsets in addition to translocation of phosphorylated STAT3 within 30 min after IL-23 stimulation. After IL-23R activation, a small subset of PKM2 also translocates to the nucleus and may contribute to STAT3 phosphorylation, suggesting multiple cellular responses including metabolic adaptation.
authors.affiliation: Kennedy Institute, Nuffield Department of Orthopedics Research Medical Science, Roosevelt Drive, Oxford OX3 7LF, UK.|Ludwig Institute for Cancer Research Ltd, Nuffield Department of Medicine, University of Oxford, Old Road Campus Research Building, Oxford OX3 7DQ, UK.
authors.forename: Corinne
authors.initials: C
authors.lastname: Lochmatter
authors.affiliation: Kennedy Institute, Nuffield Department of Orthopedics Research Medical Science, Roosevelt Drive, Oxford OX3 7LF, UK.|Target Discovery Institute, Nuffield Department of Medicine, University of Oxford, Roosevelt Drive, Oxford OX3 7FZ, UK.
authors.forename: Roman
authors.initials: R
authors.lastname: Fischer
authors.affiliation: Target Discovery Institute, Nuffield Department of Medicine, University of Oxford, Roosevelt Drive, Oxford OX3 7FZ, UK.
authors.forename: Philip D
authors.initials: PD
authors.lastname: Charles
authors.affiliation: Target Discovery Institute, Nuffield Department of Medicine, University of Oxford, Roosevelt Drive, Oxford OX3 7FZ, UK.
authors.forename: Zhanru
authors.initials: Z
authors.lastname: Yu
authors.affiliation: Kennedy Institute, Nuffield Department of Orthopedics Research Medical Science, Roosevelt Drive, Oxford OX3 7LF, UK.
authors.forename: Fiona
authors.initials: F
authors.lastname: Powrie
authors.affiliation: Target Discovery Institute, Nuffield Department of Medicine, University of Oxford, Roosevelt Drive, Oxford OX3 7FZ, UK.
authors.forename: Benedikt M
authors.initials: BM
authors.lastname: Kessler
mesh_terms: D021581:Active Transport, Cell Nucleus; D000220:Adaptation, Biological; D002352:Carrier Proteins / Q000378:metabolism; D030562:Databases, Protein; D004734:Energy Metabolism; D006019:Glycolysis; D006801:Humans; D053759:Interleukin-23 / Q000494:pharmacology; D008214:Lymphocytes / Q000378:metabolism*; D008565:Membrane Proteins / Q000378:metabolism; D008954:Models, Biological; D010748:Phosphopeptides / Q000378:metabolism; D010750:Phosphoproteins / Q000378:metabolism*; D010766:Phosphorylation; D040901:Proteomics / Q000379:methods; D018123:Receptors, Interleukin / Q000378:metabolism*; D050796:STAT3 Transcription Factor / Q000378:metabolism; D015398:Signal Transduction / Q000187:drug effects; D013963:Thyroid Hormones / Q000378:metabolism; D000097810:Thyroid Hormone-Binding Proteins
chemical_list: D002352:Carrier Proteins; D053759:Interleukin-23; D008565:Membrane Proteins; D010748:Phosphopeptides; D010750:Phosphoproteins; D018123:Receptors, Interleukin; D050796:STAT3 Transcription Factor; D013963:Thyroid Hormones; D000097810:Thyroid Hormone-Binding Proteins; C515500:IL23R protein, human