MONOCLONAL ANTIBODY AGAINST A MULTIFUNCTIONAL CALMODULIN-DEPENDENT PROTEIN KINASE FROM RAT BRAIN AND THE TISSUE DISTRIBUTION OF THE ENZYME
MONOCLONAL ANTIBODY AGAINST A MULTIFUNCTIONAL CALMODULIN-DEPENDENT PROTEIN KINASE FROM RAT BRAIN AND THE TISSUE DISTRIBUTION OF THE ENZYME
复制标题
大鼠脑内多功能钙调蛋白依赖性蛋白激酶的单克隆抗体及其组织分布
DOI：
10.2220/biomedres.7.405
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发表时间：
1986
影响因子：
1.2
通讯作者：
E. Miyamoto
中科院分区：
文献类型：
--
作者：
K. Fukunaga;Hiroshi Sato;K. Takatsu;A. Tominaga;E. Miyamoto
我们已经制备了杂交瘤细胞，可以产生抗大鼠脑内多功能钙调蛋白依赖蛋白激酶(CaM-Kinase)的单抗。该单抗为IgM类，与酶的49K亚基发生交叉反应，而与60K亚基不发生交叉反应。与脑中钙调素依赖的磷酸二酯酶、钙调神经磷酸酶和其他钙调蛋白结合蛋白无交叉反应。用该单抗研究钙调蛋白-激酶在脑内的区域分布。所有脑区均检测到49K亚基，但主要分布在大脑皮层、海马体和纹状体。免疫印迹进一步分析大鼠各种组织，发现在脾、胃、胰腺、肺和脂肪组织中存在M，52,000的免疫反应蛋白，在骨骼肌和心脏中存在M，55,000的免疫反应蛋白。此外，免疫反应结果与酶活性测定结果一致。结果表明，钙调蛋白-激酶在组织中广泛分布。多功能钙调蛋白依赖的蛋白激酶(CaM-K)在神经功能中起着重要作用，如刺激-分泌偶联、神经递质的生物合成和代谢反应等。许多特定底物蛋白的存在进一步表明了该酶的生物学意义。这些包括突触素I(2，17)，微管蛋白(8)，微管相关蛋白(MAP)(4，34，35，37)，酪氨酸3-单加氧酶(36)，色氨酸52，H.佐藤博士从熊本大学医学院内科第一科休假。缩写：‘on.’SDS-PAGE，十二烷基硫酸钠-聚丙烯酰胺凝胶电泳；DTT，二硫苏糖醇单加氧酶(38)，磷酸二酯酶(5)，糖原合成酶(13，14)，脑肌球蛋白(30)，突触体内源性蛋白(6)和高迁移基染色质蛋白(28)。最近，在肝脏(1，26)、骨骼肌(33)、胰腺(10)、垂体前叶(12)、心脏(15，16，18)和肺(29)中发现了一种多功能的钙调蛋白依赖蛋白激酶，其性质与脑酶相似。事实上，来自骨骼肌的酶表现出与脑酶(21，32)相同的底物特异性和免疫学特性。在本研究中，我们制备了一株产生抗脑钙调蛋白激酶的单抗的杂交瘤，该抗体属于IgM类。用该单抗研究了与脑酶相似的酶的组织分布。。。
We have prepared hybridoma cells that produce monoclonal antibody against a multifunctional calmodulin-dependent protein kinase (calmodulin-kinase) from rat brain. The monoclonal antibody was of the IgM class and cross-reacted with the 49 K-subunit of the enzyme but not with the 60 K-subunit. There was no crossreaction with the Ca“-calmodulin-dependent phosphodiesterase, calcineurin and other calmodulin-binding proteins from the brain. The monoclonal antibody was used to study regional distribution of calmodulin-kinase in the brain. The 49 K subunit was detected in all brain regions examined, but was found predominantly in cerebral cortex, hippocampus and striatum. Further analysis ofvarious rat tissues by immunoblotting revealed the presence of immunoreactive proteins with M, 52,000 in the spleen, stomach, pancreas, lung and adipose tissue and with the M, 55,000 in the skeletal muscle and heart. In addition, the findings obtained by immunoreactivity were consistent with those determined by enzyme activity. The results suggest that calmodulin-kinase is widely distributed in the tissues. The multifunctional calmodulin-dependent protein kinase (calmodulin-kinase) plays an important role in the neural function, such as the stimulus-secretion coupling, biosynthesis of neurotransmitters and metabolic reactions. The biological significance of the enzyme is further suggested by the presence of many specific substrate proteins. These include synapsin I (2, 17), tubulin (8), microtubuleassociated proteins (MAPs) (4, 34, 35, 37), tyrosine 3-monooxygenase (36), tryptophan 52Dr H. Sato is on leave from the First Department of Internal Medicine, Kumamoto University Medical School. Abbreviat:'ons.' SDS-PAGE, sodium dodecyl sulfate-polyacrylamide gel electrophoresis; DTT, dithiothreitol monooxygenase (38), phosphodiesterase (5), glycogen synthase (13, 14), brain myosin (30), endogenous proteins in synaptosomes (6) and a high mobility group chromatin protein (28). Recently, a multifunctional calmodulin-dependent protein kinase that has similar properties to the brain enzyme was found in the liver (1, 26), skeletal muscle (33), pancreas (10), anterior pituitary gland (12), heart (15, 16, 18) and lung (29). In fact, the enzyme from the skeletal muscle shows substrate specificity and immunological property identical with the brain enzyme (21, 32). In the present study, we prepared a hybridoma producing monoclonal antibody against brain calmodulin-kinase, which was of the IgM class. The monoclonal antibody was used for the study on tissue distribution of the enzyme similar to the brain enzyme. ..