A STUDY OF PROTEINS WITHIN THE CNS BY TWO-DIMENSIONAL GEL ELECTROPHORESIS

在中枢神经系统内利用蛋白质的鉴定 2-双向凝胶电泳（2DE）继续进行。 1)之一 我们大脑凝胶上最大的蛋白质，“蛋白质36”已经被分离出来。 并从大鼠脑匀浆中纯化。 蛋白质36是一种 分子量为64 kD的同二聚体和单体 分子量为37 kD，pI为6.5。 免疫细胞化学研究 使用针对这种蛋白质的兔子抗体， 36位于大锥体细胞、树突和轴突中， 大脑皮层的第五层 2)第二种蛋白质（“蛋白质94”） 已经从牛脑中分离并纯化。 这是一 cAMP刺激的磷蛋白（M.W. 94 kD）， 最初在大鼠新纹状体中发现。 纯化 通过32 P-磷酸化样品的放射自显影监测 通过SDS-PAGE分离。 免疫细胞化学研究显示 94蛋白定位于星形胶质细胞 3)我们检测到 45 Ca ~（++）在大鼠大脑皮层中至少有3种Ca ~（++）结合蛋白 放射自显影 两种蛋白质被鉴定为钙调素， 钙调磷酸酶B亚单位。 此外，一种未鉴定的Ca++ - 分子量约18 kD，pI 5.4的结合蛋白。 揭示 4)七种高亲和力钙结合蛋白具有 在硫酸铵沉淀后检测到。 三 的蛋白质已被确定（钙调蛋白，B亚基的 钙调神经磷酸酶和维生素D依赖性钙结合蛋白。 其他4种蛋白质的身份尚不清楚。 5)我们有 研究了存在于七个离散的显微解剖的 大脑区域 区域差异明显， 在基础条件下研究的脑内磷酸化 环腺苷酸依赖性蛋白激酶激活后 和钙/钙调蛋白依赖性蛋白激酶C。 6)一系列检查听觉核内蛋白质的研究 正在进行中。 结果表明， 在二维凝胶上可见的蛋白质穿过听觉核， 更普遍的是在感觉核和其他大脑区域之间。

The identification of interesting proteins within the CNS utilizing 2-dimensional gel electrophoresis (2DE) continues. 1) One of the largest proteins on our brain gels, "protein 36" has been isolated and purified from homogenates of rat brain. Protein 36 is a homodimer with a molecular weight of 64 kD and a monomeric weight of 37 kD with a pI of 6.5. Immunocytochemical studies using a rabbit antibody raised to this protein revealed that protein 36 is localized in large pyramidal cells, dendrites and axons of layer V of the cerebral cortex. 2) A second protein ("protein 94") has been isolated from the bovine brain and purified. This is a cAMP-stimulated phosphoprotein (M.W. 94 kD) which was originally identified in the rat neostriatum. Purification was monitored by autoradiography of 32P-phosphorylated samples separated by SDS-PAGE. Immunocytochemical studies revealed that protein 94 was localized in astrocytes. 3) We have detected at least 3 Ca++ -binding proteins in the rat cortex by 45Ca++ autoradiography. Two proteins were identified as calmodulin and the B subunit of calcineurin. In addition, an unidentified Ca++ - binding protein of molecular weight about 18 kD and pI 5.4 was revealed. 4) Seven high affinity calcium binding proteins have been detected following ammonium sulfate precipitation. Three of the proteins have been identified (calmodulin, B subunit of calcineurin and vitamin D-dependent calcium binding protein. The identity of the other 4 proteins are unknown. 5) We have studied phosphoproteins present in seven discrete microdissected brain areas. Marked regional differences in the pattern of phosphorylation among the brain studied under basal conditions and following activation of cyclic AMP-dependent protein kinase and calcium/calmodulin-dependent protein kinase C were noted. 6) A series of studies examining proteins within auditory nuclei are under progress. Results indicate several differences in proteins visible on two-dimensional gels across auditory nuclei or more generally between sensory nuclei and other brain areas.