PROTEIN PHOSPHORYLATION, EPILEPSY & ANTICONVULSANTS

该实验室的研究表明，苯妥英， 克拉马西平和苯二氮卓类(BZ)对钙调素的抑制作用 调节的蛋白质磷酸化和神经递质释放 突触准备。根据这些结果提出了一个假设。 他说，钙离子调节特定大脑蛋白质的磷酸化， 特别是在突触，可能在调节中发挥重要作用 神经元兴奋性和特定抗惊厥药物的作用。 这项调查将系统地检验这一假设。首字母 研究将针对分离Ca++钙调蛋白激酶系统和 钙离子、钙调蛋白和抗惊厥药物的作用特征 化合物对蛋白质磷酸化、神经递质释放和 钙离子在各种突触准备中的调节作用。我们会 几种苯二氮卓类药物抑制活性的相关性 这些钙离子调节事件及其抑制最大值的效力 电击引起的抽搐。这个实验室的结果是 在脑膜上发现了一个新的抗惊厥药物结合部位。这个 苯二氮卓类药物与这个部位结合的效力非常相关 显著地具有抑制最大电击诱导的能力 癫痫发作与Ca++钙调蛋白激酶活性的关系。这些结果提供了 脑内存在抗惊厥药物结合部位的证据 可能调节Ca++钙调蛋白激酶活性的膜 系统和调节突触膜的兴奋性。用人 高纯度钙调素的光亲和标记实验 ，我们将能够确定一个亚基或 激酶系统的调节剂是膜抗惊厥剂的一部分 结合部位。还将开始研究一种化合物的特征 最近从大脑中分离出的一种有效的钙离子抑制物 钙调蛋白激酶系统。我们的初步研究表明，这一新的 化合物也是一种有效的抗惊厥药物，可以抑制最大电流。 电击导致癫痫发作。拟议的实验将使我们能够 鉴定这种新的内源性抗惊厥物质，并确定 它是癫痫发作时从大脑中释放出来的一种“天然”抗惊厥剂 对癫痫发作活动的抑制。

Studies from this laboratory have demonstrated that phenytoin, crbamazepine, and the benzodiazepines (BZ) inhibit Ca++ calmodulin regulated protein phosphorylation and neurotransmitter release in several synaptic preparations. An hypothesis was proposed from these results stating that the Ca++regulated phosphorylation of specific brain proteins, especially at the synapse, may play an important role in regulating neuronal excitability and the action of specific anticonvulsant drugs. This investigation will systematically examine this hypothesis. Initial studies will be directed at isolating the Ca++ calmodulin kinase system and characterizing the effects of Ca++, calmodulin, and anticonvulsant compounds on protein phosphorylation, neurotransmitter release, and Ca++regulated functions in various synaptic preparations. We will correlate the potency of several benzodiazepine derivatives to inhibit these Ca++ regulated events with their potency for inhibiting maximal electric shock induced convulsions. Results from this laboratory have identified a novel anticonvulsant binding site in brain membrane. The potency of the benzodiazepines to bind to this site correlate very significantly with their ability to inhibit maximal electric shock induced seizures and Ca++ calmodulin kinases activity. These results provide evidence for the existence of an anticonvulsant binding site in brain membrane that may regulate the activity of the Ca++ calmodulin kinase system and modulate the excitability of synaptic membrane. Employing photoaffinity labelling experiments with highly purified Ca++ calmodulin kinase and membrane fractions, we will be able to determine if a subunit or modulator of the kinase system is part of the membrane anticonvulsant binding site. Studies will also be initiated to characterize a compound recently isolated from brain that is a potent inhibitor of the Ca++ calmodulin kinase system. Our preliminary studies indicate that this new compound is also a potent anticonvulsant in inhibiting maximal electric shock induced seizures. The proposed experiments will allow us to characterize this new endogenous anticonvulsant substance and determine if it is a "natural" anticonvulsant released from brain during seizure activity with the resultant suppression of seizure activity.