Galve-Roperh I, Malpartida J M, Haro A, Diaz-Laviada I
Departamento de Bioquímica y Biología Molecular I, Facultad de Químicas, Universidad Complutense, Madrid, Spain.
Neurosci Lett. 1996 Nov 15;219(1):68-70. doi: 10.1016/s0304-3940(96)13165-7.
Phosphatidylcholine breakdown has been shown to play a critical role in signal transduction involving generation of a number of second messengers [Exton, J.H., Biochim. Biophys. Acta, 1212 (1994) 26-42]. In the present report we demonstrate by immunofluorescence that short-treatment of C 6 glial cells with phosphatidylcholine-hydrolyzing phospholipase C (PC-PLC), changes the intracellular localization of protein kinase C (PKC) zeta from the cytoplasm to a perinuclear region. Western blot analysis also showed a redistribution of PKC zeta after incubation of cells with PC-PLC. To test whether these changes were accompanied by an activation of the enzyme, we measured the extent of phosphorylation of PKC zeta by immunoprecipitation from 32P-labelled cells. Short-treatment with PC-PLC resulted in enhanced phosphorylation of the higher Mr PKC zeta in C 6 glial cells.
磷脂酰胆碱的分解在涉及多种第二信使生成的信号转导中发挥着关键作用[埃克斯顿，J.H.，《生物化学与生物物理学学报》，1212（1994）26 - 42]。在本报告中，我们通过免疫荧光证明，用磷脂酰胆碱水解磷脂酶C（PC - PLC）对C6神经胶质细胞进行短期处理，会使蛋白激酶C（PKC）ζ的细胞内定位从细胞质改变至核周区域。蛋白质印迹分析也显示，细胞与PC - PLC孵育后PKCζ发生了重新分布。为了测试这些变化是否伴随着该酶的激活，我们通过从32P标记的细胞中进行免疫沉淀来测量PKCζ的磷酸化程度。用PC - PLC进行短期处理导致C6神经胶质细胞中较高分子量的PKCζ磷酸化增强。
