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首页> 外文期刊>Journal of Neurochemistry: Offical Journal of the International Society for Neurochemistry >Identification of the isoforms of Ca(2+)/Calmodulin-dependent protein kinase II in rat astrocytes and their subcellular localization.
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Identification of the isoforms of Ca(2+)/Calmodulin-dependent protein kinase II in rat astrocytes and their subcellular localization.

机译：Ca（2 +）/钙调蛋白依赖性蛋白激酶II在大鼠星形胶质细胞中的同工型及其亚细胞定位的鉴定。

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摘要

Ca(2+)/calmodulin-dependent protein kinase II (CaM kinase II) occurs in astrocytes as well as in neurons in brain. We have reported that CaM kinase II is involved in the regulation of cytoskeletal proteins and gene expression in astrocytes. In this study, we identified all isoforms of CaM kinase II in astrocytes and examined their subcellular localization. When we amplified the isoforms of four subunits by RT-PCR followed by the "nested" PCR, totally 10 isoforms were obtained. Immunoblot analyses with five types of antibodies against CaM kinase II indicated that the most abundant isoform was delta2. Immunostaining suggested that the delta2 isoform was localized predominantly at the Golgi apparatus. The localization of the delta2 isoform at the Golgi apparatus was also observed in NG108-15 cells. We overexpressed all isoforms that contained the nuclear localization signal to examine their nuclear targeting in NG108-15 cells. In contrast to the alphaB and delta3 isoforms that entered the nucleus, as reported, the gammaA isoform was excluded from the nucleus in the transfected NG108-15 cells. These results suggest that the 15-amino acid insertion following the nuclear localization signal inhibits the nuclear targeting of the gammaA isoform.
机译：Ca（2 +）/钙调蛋白依赖性蛋白激酶II（CaM激酶II）发生在星形胶质细胞以及大脑的神经元中。我们已经报道CaM激酶II参与星形胶质细胞中细胞骨架蛋白和基因表达的调节。在这项研究中，我们鉴定了星形胶质细胞中CaM激酶II的所有同工型，并检查了其亚细胞定位。当我们通过RT-PCR然后“嵌套” PCR扩增四个亚基的同工型时，总共获得了10个同工型。用五种针对CaM激酶II的抗体进行的免疫印迹分析表明，最丰富的同工型为delta2。免疫染色提示δ2同工型主要位于高尔基体。在NG108-15细胞中也观察到了delta2亚型在高尔基体的定位。我们过表达包含核定位信号的所有同工型，以检查其在NG108-15细胞中的核靶向。与报道的进入细胞核的alphaB和delta3同工型相反，在转染的NG108-15细胞中，γA同工型被排除在细胞核之外。这些结果表明，在核定位信号后插入15个氨基酸会抑制gammaA同种型的核靶向。

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