Protein tyrosine phosphorylation associated with activation of the interleukin 2 receptor.
The addition of interleukin 2 (IL2) to the IL2-dependent murine cytotoxic T cell line CTTL-2 induced increased tyrosine phosphorylation of a protein with a molecular weight of 80,000 and, to a lesser extent, proteins with molecular weights of 130,000, 100,000, and 69,000. To correlate the stimulation of tyrosine phosphorylation with increased tyrosine kinase activity, cell-free phosphorylation assays were performed. Phosphotyrosine-containing proteins were purified from detergent-solubilized cell lysates by immunoprecipitation with anti-phosphotyrosine antibodies. The level of tyrosine kinase activity was determined by incorporation of [gamma-32P]ATP into the exogenous substrate histone H2B. IL2 treatment of cells increased H2B phosphorylation 10-fold when compared with nonstimulated cells. Phosphorylation was first detected after 2.5 min of incubation with physiologically relevant (100 pM) IL2 doses. To examine if tyrosine kinase activity was resident within the IL2 receptor complex, cell-free phosphorylation assays were performed with ligand-receptor complexes following cross-linking with IL2 and purification by immunoabsorption with an anti-IL2 antibody. Tyrosine kinase activity was found specifically associated with the IL2 receptor complex. These results indicate that the IL2 receptor complex contains a tyrosine kinase activity that is induced by IL2 binding and suggest that components of the complex may be a substrate of this activity.
Department of Pathology and Laboratory Medicine, University of Pennsylvania, School of Medicine, Philadelphia 19104.
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D000255:Adenosine Triphosphate / Q000378:metabolism; D000818:Animals; D002460:Cell Line; D004591:Electrophoresis, Polyacrylamide Gel; D004789:Enzyme Activation; D006657:Histones / Q000378:metabolism; D007163:Immunosorbent Techniques; D007376:Interleukin-2 / Q000494:pharmacology*; D051379:Mice; D008970:Molecular Weight; D010710:Phosphates / Q000378:metabolism; D010766:Phosphorylation; D019000:Phosphotyrosine; D011505:Protein-Tyrosine Kinases / Q000378:metabolism*; D015375:Receptors, Interleukin-2 / Q000187:drug effects / Q000378:metabolism*; D014443:Tyrosine / Q000031:analogs & derivatives* / Q000378:metabolism
D006657:Histones; D007376:Interleukin-2; D010710:Phosphates; D015375:Receptors, Interleukin-2; D019000:Phosphotyrosine; D014443:Tyrosine; D000255:Adenosine Triphosphate; D011505:Protein-Tyrosine Kinases