Dynamic Interaction- and Phospho-Proteomics Reveal Lck as a Major Signaling Hub of CD147 in T Cells.
Numerous publications have addressed CD147 as a tumor marker and regulator of cytoskeleton, cell growth, stress response, or immune cell function; however, the molecular functionality of CD147 remains incompletely understood. Using affinity purification, mass spectrometry, and phosphopeptide enrichment of isotope-labeled peptides, we examined the dynamic of the CD147 microenvironment and the CD147-dependent phosphoproteome in the Jurkat T cell line upon treatment with T cell stimulating agents. We identified novel dynamic interaction partners of CD147 such as CD45, CD47, GNAI2, Lck, RAP1B, and VAT1 and, furthermore, found 76 CD147-dependent phosphorylation sites on 57 proteins. Using the STRING protein network database, a network between the CD147 microenvironment and the CD147-dependent phosphoproteins was generated and led to the identification of key signaling hubs around the G proteins RAP1B and GNB1, the kinases PKCβ, PAK2, Lck, and CDK1, and the chaperone HSPA5. Gene ontology biological process term analysis revealed that wound healing-, cytoskeleton-, immune system-, stress response-, phosphorylation- and protein modification-, defense response to virus-, and TNF production-associated terms are enriched within the microenvironment and the phosphoproteins of CD147. With the generated signaling network and gene ontology biological process term grouping, we identify potential signaling routes of CD147 affecting T cell growth and function.
Molecular Immunology Unit, Institute for Hygiene and Applied Immunology, Center of Pathophysiology, Infectiology and Immunology, Medical University of Vienna, 1090 Vienna, Austria; and.
Verena
0000-0003-2831-600X
V
Supper
Molecular Immunology Unit, Institute for Hygiene and Applied Immunology, Center of Pathophysiology, Infectiology and Immunology, Medical University of Vienna, 1090 Vienna, Austria; and.
Ingrid
I
Hartl
Microchemistry Core Facility, Max Planck Institute of Biochemistry, 82152 Martinsried, Germany.
Cyril
C
Boulègue
Molecular Immunology Unit, Institute for Hygiene and Applied Immunology, Center of Pathophysiology, Infectiology and Immunology, Medical University of Vienna, 1090 Vienna, Austria; and.
Anna
0000-0002-8005-8522
A
Ohradanova-Repic
Molecular Immunology Unit, Institute for Hygiene and Applied Immunology, Center of Pathophysiology, Infectiology and Immunology, Medical University of Vienna, 1090 Vienna, Austria; and hannes.stockinger@meduniwien.ac.at.
Hannes
0000-0001-6404-4430
H
Stockinger
D051926:Basigin / Q000235:genetics / Q000378:metabolism*; D014408:Biomarkers, Tumor / Q000378:metabolism*; D048708:Cell Growth Processes; D003599:Cytoskeleton / Q000378:metabolism*; D000091342:Endoplasmic Reticulum Chaperone BiP; D019204:GTP-Binding Proteins / Q000378:metabolism; D063990:Gene Ontology; D018014:Gene Transfer Techniques; D006801:Humans; D019169:Jurkat Cells; D008213:Lymphocyte Activation; D019860:Lymphocyte Specific Protein Tyrosine Kinase p56(lck) / Q000378:metabolism*; D046912:Multiprotein Complexes / Q000378:metabolism*; D010766:Phosphorylation; D011485:Protein Binding; D040901:Proteomics; D015398:Signal Transduction; D013312:Stress, Physiological; D013601:T-Lymphocytes / Q000502:physiology*
D014408:Biomarkers, Tumor; D000091342:Endoplasmic Reticulum Chaperone BiP; C000717247:HSPA5 protein, human; D046912:Multiprotein Complexes; D051926:Basigin; D019860:Lymphocyte Specific Protein Tyrosine Kinase p56(lck); D019204:GTP-Binding Proteins