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研究佛波酯(PMA)对人肝癌细胞7721胞液和膜性组分中3类蛋白激酶(TPK、PKA、PKC)的早期(5~60min)效应。采用体外细胞培养和蛋白激酶的同位素标记底物测定法。结果:PMA在30min内可促进PKA由膜向胞液的转位和PKC由胞液向膜的转位,并能迅速促进胞液PKC的下降,但膜性PKC的下降发生在30min的活力高峰以后。60min时膜性PKA和PKC都恢复到正常水平。PMA还使胞液TPK持续升高,高峰在5min,而使膜性TPK先降低,40min后才明显升高。结论:PMA对蛋白激酶的早期作用相当多样化,不单是通过受体PKC起作用。
To study the early (5-60 min) effect of phorbol ester (PMA) on three protein kinases (TPK, PKA, PKC) in the cytosol and membranous components of human hepatoma cell 7721. In vitro cell culture and isotopically labeled substrate assays for protein kinases were used. RESULTS: PMA promoted the translocation of PKA from the membrane to the cytosol and the translocation of PKC from the cytosol to the membrane within 30 min, and could rapidly promote the decrease of cytosolic PKC, but the decrease of membrane PKC occurred at the peak of 30 min. after. At 60 minutes, both membrane PKA and PKC returned to normal levels. PMA also increased the cytoplasmic TPK continuously, with the peak at 5 min, while the membrane TPK decreased first, and then increased significantly after 40 min. Conclusion: The early effects of PMA on protein kinases are quite diverse, not only through the receptor PKC.