Characterization of interleukin 2-stimulated phosphorylation of 67 and 63 kDa proteins in human T-cells.
We have investigated rapid and marked phosphorylation of cellular proteins induced by interleukin 2 (IL-2) in both phytohaemagglutinin-stimulated normal peripheral blood leucocytes, and IL-2-dependent or -independent human T-cell lines bearing human T-cell leukaemia (lymphotropic) virus type I. Two-dimensional electrophoretic analysis showed that the IL-2-induced phosphoprotein was of Mr 67,000 with a pI of 5.8 (pp67) and was distinct from the IL-2 receptor. IL-2 also stimulated phosphorylation of four other proteins, with an Mr of 63,000 and pI values 5.3-6.1 (pp63s). The stimulation of pp67 phosphorylation was observed within 5 min after addition of IL-2 and was maximal after 15 min. The maximal phosphorylation was more than 10-fold that observed initially. In IL-2-dependent cells, IL-2 dose responses of pp67 phosphorylation and cell proliferation were exactly correlated. Phosphoamino acid analysis showed that the phosphorylation site of pp67 and pp63s was a serine residue. Subcellular-fractionation studies indicated that pp67 was localized in cytosol, whereas pp63s phosphorylation was induced by IL-2 in nuclear and cytosol fractions. Similar phosphorylation of pp67 and pp63s was observed when the cells were treated with phorbol 12-myristate 13-acetate instead of IL-2. These results suggest that IL-2-IL-2-receptor interaction leads to activation of protein kinase(s), resulting in phosphorylation of certain cellular proteins such as pp67 and pp63s, and that this phosphorylation could be an early event in the transmission of intracellular growth signalling from the IL-2 receptors.
D002455:Cell Division / Q000187:drug effects; D002460:Cell Line; D004305:Dose-Response Relationship, Drug; D004591:Electrophoresis, Polyacrylamide Gel; D006801:Humans; D007376:Interleukin-2 / Q000494:pharmacology*; D007700:Kinetics; D010766:Phosphorylation; D011506:Proteins / Q000378:metabolism*; D013268:Stimulation, Chemical; D013601:T-Lymphocytes / Q000187:drug effects / Q000378:metabolism*; D013755:Tetradecanoylphorbol Acetate / Q000494:pharmacology
D007376:Interleukin-2; D011506:Proteins; D013755:Tetradecanoylphorbol Acetate