ROLE OF PROTEIN-KINASE-C IN THE REGULATION OF GAP JUNCTIONAL COMMUNICATION
ROLE OF PROTEIN-KINASE-C IN THE REGULATION OF GAP JUNCTIONAL COMMUNICATION
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DOI：
10.1002/tcm.1770140603
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发表时间：
1994-01-01
期刊：
影响因子：
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通讯作者：
MILOSZEWSKA, J
中科院分区：
文献类型：
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作者：
BUDUNOVA, IV;MITTELMAN, LA;MILOSZEWSKA, J
我们研究了 SV40 转化的 Djungarian 仓鼠成纤维细胞（DM15 细胞）中蛋白激酶 (PKC) 消耗对间隙连接通透性、Cx43 电泳迁移率和细胞对不同解偶联刺激的敏感性水平的影响。暴露于 12-O-十四烷酰佛波醇-13-乙酸酯 (TPA) 24 小时后，与完整细胞相比，DM15 细胞中的总 PKC 活性降低至 20-25%。在 PKC 耗尽的细胞中，染料偶联水平比相同的未处理培养物高 30-40%。蛋白质印迹分析揭示了间隙连接蛋白连接蛋白 43 的多种形式，对应于该蛋白已知的磷酸化和去磷酸化形式。 PKC 耗尽后，未观察到连接蛋白 43 磷酸化水平降低。 TPA(10(-7) g/ml)、mezerein (10(-7) g/ml)、teleocidin (10(-8) g/ml)、钙离子载体 A23I87 (10(-6) g/ml)、杀虫剂 1,1,1-三氯-2,2-双-(对氯苯基)-(DDT) (10(-4) g/ml) 和尼日利亚菌素(10(-5) M，水解物乳清蛋白溶液，pH 6.3) 导致染料偶联测定中受体细胞数量减少四到六倍。 PKC 耗尽的细胞几乎完全抵抗 mezerein、teleocidin 和 A23187 的解偶联作用，以及新接触的 TPA，并且部分抵抗 DDT 的作用。尼日利亚菌素抑制 PKC 耗尽的细胞之间的细胞间通讯的程度与对照细胞之间的相同。因此，在所研究的细胞系统中，PKC在维持间隙连接通透性的基础水平方面发挥一定作用，并且作为TPA、mezerein、teleocidin和Ca2+等物质的解偶联作用的介体具有重要意义。 (C) 1994 年 Wiley-Liss 公司
We have examined the effect of protein kinase (PKC) depletion in SV40-transformed Djungarian hamster fibroblasts (DM15 cells) on the level of gap junction permeability, Cx43 electrophoretic mobility, and cell sensitivity to different uncoupling stimuli. After 24 hr exposure to 12-O-tetradecanoyl-phorbol-13-acetate (TPA), the total PKC activity in DM15 cells was reduced to 20-25% in comparison with intact cells. In PKC-depleted cells the level of dye coupling was 30-40% higher than in the same untreated cultures. Western blot analysis revealed multiple forms of the gap junction protein connexin 43, which correspond to known phosphorylated and dephosphorylated forms of this protein. No decrease in the level of connexin 43 phosphorylation after PKC depletion was observed. TPA(10(-7) g/ml), mezerein (10(-7) g/ml), teleocidin (10(-8) g/ml), Ca-ionophore A23I87 (10(-6) g/ml), insecticide 1,1,1-trichloro-2,2-bis-(p-chlorphenyl)-(DDT) (10(-4) g/ml), and nigericin (10(-5) M in hydrolysate lactalbumin solution, pH 6.3) induced a four-to six-fold decrease in the number of recipient cells in the dye-coupling assay. PKC-depleted cells became almost completely resistant to the uncoupling effect of mezerein, teleocidin, and A23187, as well as to new exposure to TPA, and became partially resistant to the effect of DDT. Nigericin inhibited intercellular communication between PKC-depleted cells to the same extent as between control cells. Thus, in the cell system studied, PKC plays a certain role in maintaining the basal level of gap junction permeability and has an important significance as a mediator of the uncoupling effects of such substances as TPA, mezerein, teleocidin, and Ca2+. (C) 1994 Wiley-Liss, Inc.