Visualization of protein kinases in lymphocytes stimulated to proliferate with concanavalin A or inhibited with a phorbol ester |
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Authors: | A P Iyer S A Pishak M J Sniezek A M Mastro |
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Affiliation: | Microbiology Program, Department of Biochemistry, Microbiology, Molecular and Cell Biology, The Pennsylvania State University, University Park, Pennsylvania 16802 USA |
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Abstract: | Stimulation of lymphocytes with a mitogenic lectin such as concanavalin A (ConA) results in differentiation and cell division. Among the changes which occur after stimulation are increases in phosphorylation of proteins and in protein kinase activity. We used a high-resolution, nondenaturing gel system to separate and visualize protein kinases in situ. We have clearly identified both autophosphorylating and substrate-dependent kinases. One band of cyclic AMP-dependent kinase activity was significantly enhanced in lectin-stimulated cells. In contrast, treatment of the cells with phorbol ester under conditions which depress stimulation caused a decrease in the activity of one kinase. |
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Keywords: | ConA Concanavalin A kDa kilodalton PBS phosphate-buffered saline PMSF phenylmethylsulfonyl fluoride TPA 12-0-tetradecanoyl phorbol-13-acetate |
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