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Visualization of protein kinases in lymphocytes stimulated to proliferate with concanavalin A or inhibited with a phorbol ester
Authors:A P Iyer  S A Pishak  M J Sniezek  A M Mastro
Affiliation:Microbiology Program, Department of Biochemistry, Microbiology, Molecular and Cell Biology, The Pennsylvania State University, University Park, Pennsylvania 16802 USA
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.
Keywords:ConA  Concanavalin A  kDa  kilodalton  PBS  phosphate-buffered saline  PMSF  phenylmethylsulfonyl fluoride  TPA  12-0-tetradecanoyl phorbol-13-acetate
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