Ionic responses rapidly elicited by activation of protein kinase C in quiescent Swiss 3T3 cells.
Ionic responses rapidly elicited by activation of protein kinase C in quiescent Swiss 3T3 cells.
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在静止的 Swiss 3T3 细胞中，蛋白激酶 C 的激活可快速引发离子反应。
DOI：
10.1073/pnas.82.8.2384
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发表时间：
1985
影响因子：
11.1
通讯作者：
E. Rozengurt
中科院分区：
文献类型：
--
作者：
F. Vara;J. Schneider;E. Rozengurt
二酰基甘油和佛波酯激活完整细胞中的蛋白激酶C。我们在这里报道，将合成的二酰基甘油1-油基-2-乙酰甘油（OAG）添加到静止培养的瑞士3T3细胞中，可以显著提高瓦阿因敏感的86Rb+摄取率，这是Na+/K+泵活性的一个衡量指标。该效应呈剂量依赖性，暴露于二酰基甘油1分钟后即可检测到。OAG通过阿米洛利敏感途径刺激Na+内流，使细胞内pH升高0.15 pH单位。Phorbol 12,13-二丁酸酯（PBt2）也增强了瓦他因敏感的86Rb+摄取和阿米洛利敏感的22Na+内流。饱和剂量PBt2延长3T3细胞处理时间（40小时），减少PBt2结合位点的数量和蛋白激酶C活性，消除细胞对随后添加OAG或PBt2的离子反应。使用酸“负荷”和Na+离子载体莫能菌素进行适当控制表明，在pbt2脱敏细胞中，Na+/H+反转运系统和Na+/K+泵的功能未受到损害。我们认为，蛋白激酶C的激活直接或间接地引起Na+/H+反端口活性的增强，进而导致Na+内流、细胞内pH调节和Na+/K+泵的刺激。
Diacylglycerol and phorbol esters activate protein kinase C in intact cells. We report here that addition of the synthetic diacylglycerol 1-oleoyl-2-acetylglycerol (OAG) to quiescent cultures of Swiss 3T3 cells caused a marked increase in the rate of ouabain-sensitive 86Rb+ uptake, a measure of the activity of the Na+/K+ pump. The effect was dose-dependent and could be detected after 1 min of exposure to the diacylglycerol. OAG stimulated Na+ influx via an amiloride-sensitive pathway and increased intracellular pH by 0.15 pH unit. Phorbol 12,13-dibutyrate (PBt2) also enhanced ouabain-sensitive 86Rb+ uptake and amiloride-sensitive 22Na+ influx. Prolonged treatment (40 hr) of 3T3 cells with PBt2 at a saturating dose, which reduces the number of PBt2 binding sites and protein kinase C activity, abolished the ionic response of the cells to a subsequent addition of either OAG or PBt2. Appropriate controls using acid "loads" and the Na+ ionophore monensin showed that the function of the Na+/H+ antiport system and of the Na+/K+ pump was not impaired in the PBt2-desensitized cells. We suggest that activation of protein kinase C elicits, either directly or indirectly, enhanced Na+/H+ antiport activity, which, in turn, leads to Na+ influx, intracellular pH modulation, and stimulation of the Na+/K+ pump.