Bronstein J, Nishimura R, Lasher R, Cole R, de Vellis J, Farber D, Wasterlain C
Department of Neuroscience, UCLA School of Medicine.
J Neurochem. 1988 Jan;50(1):45-9. doi: 10.1111/j.1471-4159.1988.tb13227.x.
Calcium- and calmodulin-dependent protein kinase activity was studied in pure neuronal and glial cultures. The addition of calcium and calmodulin stimulated 32P incorporation into several neuronal proteins including two in the 50- and 60-kilodalton (kD) region which comigrated with purified forebrain calmodulin kinase II subunits (CaM kinase II). In mature astrocytes, CaM kinase activity was also present, and was inhibited by trifluoroperazine and diazepam. Again in homogenates of these cells, two phosphoproteins of apparent molecular masses of 50 and 60 kD comigrated with purified CaM kinase. CaM kinase activity was absent in immature mixed glia and oligodendrocytes. The presence of CaM kinase in neurons and mature astrocytes was confirmed using monoclonal antibodies specific for the 50-kD subunit of the enzyme. No immunoreactivity was observed in oligodendrocytes. The presence of CaM kinase in astrocytes suggests a more ubiquitous role of this enzyme in regulating cellular processes than was previously recognized.
在纯神经元和神经胶质细胞培养物中研究了钙和钙调蛋白依赖性蛋白激酶的活性。添加钙和钙调蛋白刺激了32P掺入几种神经元蛋白中，包括50和60千道尔顿（kD）区域的两种蛋白，它们与纯化的前脑钙调蛋白激酶II亚基（CaM激酶II）迁移率相同。在成熟星形胶质细胞中，也存在CaM激酶活性，并且被三氟拉嗪和地西泮抑制。同样在这些细胞的匀浆中，两种表观分子量为50和60 kD的磷蛋白与纯化的CaM激酶迁移率相同。在未成熟的混合神经胶质细胞和少突胶质细胞中不存在CaM激酶活性。使用对该酶50-kD亚基特异的单克隆抗体证实了神经元和成熟星形胶质细胞中存在CaM激酶。在少突胶质细胞中未观察到免疫反应性。星形胶质细胞中存在CaM激酶表明该酶在调节细胞过程中的作用比以前认识到的更为普遍。
