McClung J K, Kletzien R F
Biochim Biophys Acta. 1981 Sep 4;676(3):300-6. doi: 10.1016/0304-4165(81)90163-x.
In this study, we have employed native polyacrylamide gels to resolve protein kinase isozymes in cytosol samples from rat liver parenchymal cells. Five distinct bands of protein kinase activity were detected. Four of these were found to be cyclic AMP-dependent. Cytosols of isolated hepatocytes from fed and fasted rats were compared using the gel system. The resulting protein kinase activity profile revealed no marked differences in isozyme pattern or kinase activity between the two nutritional states. Isolated hepatocytes from fed rats were placed in primary culture and after 24 and 48 h cytosol samples were prepared and assayed for protein kinase activity using the gel system. The isozyme pattern remained intact for cells maintained in either Waymouth's or Swim's-77 medium for at least 48 h after inoculation into culture, and the isozyme pattern was not altered by glucocorticoid treatment.
在本研究中,我们使用天然聚丙烯酰胺凝胶来分离大鼠肝实质细胞胞质溶胶样品中的蛋白激酶同工酶。检测到五条不同的蛋白激酶活性带。其中四条被发现是依赖环磷酸腺苷的。使用该凝胶系统比较了喂食和禁食大鼠分离的肝细胞的胞质溶胶。所得的蛋白激酶活性图谱显示,两种营养状态下的同工酶模式或激酶活性没有明显差异。将喂食大鼠分离的肝细胞进行原代培养,24小时和48小时后制备胞质溶胶样品,并使用该凝胶系统测定蛋白激酶活性。接种到培养物中后,在韦茅斯培养基或斯威姆77培养基中维持至少48小时的细胞,其同工酶模式保持完整,糖皮质激素处理也未改变同工酶模式。