Angiotensin II but not potassium induces subcellular redistribution of protein kinase C in bovine adrenal glomerulosa cells.
Angiotensin II but not potassium induces subcellular redistribution of protein kinase C in bovine adrenal glomerulosa cells.
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血管紧张素 II（而非钾）可诱导牛肾上腺肾小球细胞中蛋白激酶 C 的亚细胞重新分布。
DOI：
10.1016/s0021-9258(18)47524-2
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发表时间：
1987
期刊：
影响因子：
--
通讯作者：
M. Vallotton
中科院分区：
文献类型：
--
作者：
U. Lang;M. Vallotton
钙激活，磷脂依赖性蛋白激酶（蛋白激酶C）之间的细胞质和膜组分的分布进行了检查，在牛肾上腺肾小球细胞与血管紧张素II或钾。蛋白激酶C分离从胞质溶胶和洗涤剂溶解的颗粒馏分的DEAE-纤维素色谱法。在0.05-0.09 M NaCl中洗脱出可溶性和颗粒形式的肾上腺肾小球蛋白激酶C的活性主峰。发现可溶性和颗粒形式分别占未刺激细胞中总酶活性的约95%和5%。激酶活性的第二个峰用0.15-0.19 M NaCl洗脱，其不依赖于磷脂的存在。将分离的细胞暴露于10（-8）M血管紧张素II 20分钟，导致胞质活性降低至对照值的30-40%，并导致与颗粒部分相关的蛋白激酶C活性相应增加。发现血管紧张素II的ED 50为2 nM时，血管紧张素诱导的再分布呈剂量依赖性，并且发生迅速，在5-10 min内达到平台期。特异性拮抗剂[Sar 1，Ala 8]血管紧张素II可阻止其发生。相比之下，用12 mM KCl刺激并没有改变蛋白激酶C活性的亚细胞分布。这些结果表明，蛋白激酶C的再分布代表了血管紧张素II后受体激活级联反应的早期步骤，但不是肾上腺肾小球细胞的钾刺激。
The distribution of calcium-activated, phospholipid-dependent protein kinase (protein kinase C) between cytosol and membrane fractions was examined in bovine adrenal glomerulosa cells treated with angiotensin II or potassium. Protein kinase C was isolated from cytosol and from detergent-solubilized particulate fractions by DEAE-cellulose chromatography. A major peak of activity for both the soluble and particulate forms of adrenal glomerulosa protein kinase C was eluted at 0.05-0.09 M NaCl. The soluble and particulate forms were found to constitute about 95 and 5%, respectively, of the total enzyme activity in unstimulated cells. A second peak of kinase activity was eluted with 0.15-0.19 M NaCl, which was not dependent on the presence of phospholipids. Exposure of isolated cells for 20 min to 10(-8) M angiotensin II resulted in a decrease in cytosolic activity to 30-40% of control values, and in a corresponding increase in protein kinase C activity associated with the particulate fraction. This hormone-induced redistribution was found to be dose-dependent with an ED50 of 2 nM for angiotensin II, and it occurred rapidly, reaching a plateau within 5-10 min. It was prevented by the specific antagonist [Sar1,Ala8]angiotensin II. By contrast, stimulation with 12 mM KCl did not change the subcellular distribution of protein kinase C activity. These results suggest that redistribution of protein kinase C represents an early step in the post-receptor activation cascade following angiotensin II, but not potassium stimulation of adrenal glomerulosa cells.