Inhibitory role of regucalcin in the regulation of Ca2+-dependent protein kinases activity in rat brain neurons
Inhibitory role of regucalcin in the regulation of Ca2+-dependent protein kinases activity in rat brain neurons
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DOI：
10.1016/s0022-510x(00)00476-7
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发表时间：
2001-01-15
影响因子：
4.4
通讯作者：
Yamaguchi, M
中科院分区：
文献类型：
--
作者：
Hamano, T;Yamaguchi, M
研究了调钙素对原代培养大鼠脑神经细胞蛋白激酶活性的调控作用。用细胞匀浆的5500 g上清部分检测蛋白激酶活性。在CaCl2 （10(-4) M）存在的情况下，加入钙调素（5马克杯/毫升）或二辛酸甘油（5马克杯/毫升）显著提高了蛋白激酶活性，表明神经元细胞中存在Ca2+/钙调素依赖性蛋白激酶和蛋白激酶C。在不添加钙调素或二辛烷甘油而不添加Ca2+的情况下，在酶反应混合物中添加reucalcin (10（-9)-10(-7) M）导致蛋白激酶活性显著降低。此外，在CaCl2存在的情况下，通过添加钙调素或二辛酰甘油，regulalcin完全阻止了蛋白激酶的激活（10(-4)M）。在不添加CaCl2的情况下，抗regucalcin单克隆抗体（25或50 ng/ml）的存在导致蛋白激酶活性显著升高。加入钙调素拮抗剂三氟拉嗪（2X10(-5) M）或蛋白激酶c抑制剂staurosporine （10(-6) M）可显著抑制这种作用。本研究表明，大鼠脑神经细胞内源性调钙素对Ca2+依赖性蛋白激酶活性具有抑制作用。(C) 2001 Elsevier Science B.V.版权所有
The role of regucalcin in the regulation of protein kinase activity in rat brain neuronal cells obtained from primary culture was investigated. Protein kinase activity was assayed using the 5500 g supernatant fraction of the cell homogenate. Protein kinase activity was significantly raised by the addition of calmodulin (5 mug/ml) or dioctanoylglycerol (5 mug/ml) in the presence of CaCl2 (10(-4) M), indicating that Ca2+/calmodulin-dependent protein kinase and protein kinase C is present in the neuronal cells. The addition of regucalcin (10(-9)-10(-7) M) in the enzyme reaction mixture caused a significant decrease in protein kinase activity in the absence of calmodulin or dioctanoylglycerol without Ca2+ addition. Moreover, regucalcin completely prevented the activation of protein kinase by the addition of calmodulin or dioctonoylglyceral in the presence of CaCl2 (10(-4) M). The presence of anti-regucalcin monoclonal antibody (25 or 50 ng/ml) caused a significant elevation of protein kinase activity without CaCl2, addition. Such an effect was significantly inhibited by the addition of trifluoperazine (2X10(-5) M), an antagonist of calmodulin, or staurosporine (10(-6) M), an inhibitor of protein kinase C. The present study demonstrates that endogenous regucalcin in rat brain neuronal cells has an inhibitory effect on Ca2+-dependent protein kinase activity. (C) 2001 Elsevier Science B.V. All rights reserved.