BRAIN CALCIUM AND CALMODULIN-DEPENDENT PROTEIN KINASE

脑钙和钙调蛋白依赖性蛋白激酶

• 批准号: 3397708
• 负责人: MARY B KENNEDY
• 金额: $ 22.03万
• 依托单位: CALIFORNIA INSTITUTE OF TECHNOLOGY
• 依托单位国家: 美国
• 财政年份: 1981
• 资助国家: 美国
• 起止时间: 1981-07-01 至 1992-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3397708
• 关键词: allosteric site; antibody specificity; autoradiography; brain metabolism; calcium flux; calmodulin dependent protein kinase; chemical structure function; cytoskeleton; electron microscopy; enzyme structure; gel electrophoresis; genetic manipulation; high performance liquid chromatography; hippocampus; histochemistry /cytochemistry; immunochemistry; immunological substance; laboratory mouse; laboratory rabbit; laboratory rat; microtubules; monoclonal antibody; neurotransmitters; phosphorylation; radioimmunoassay; scintillation spectrometry; synapses; synaptic vesicles

We are studying the molecular structure and function of synaptic connections between neurons in the central nervous system. Derangements in the regulation of these connections are an important part of the pathology of several neurological and mental diseases including epilepsy, Alzheimer's disease, schizophrenia, and depression. Many neurotransmitters and neurohormones regulate synaptic function by altering intracellular levels of calcium ion. We are studying the mechanisms by which these fluctuations in calcium levels alter synaptic function. We will focus on the study of a synaptic regulatory pathway that has as its central element an abundant, brain-specific calcium and calmodulin-dependent protein kinase. This kinase is a large oligomer of two distinct but homologous catalytic subunits called alpha and beta. In the forebrain, including the hippocampus, cortex and striatum, the kinase is extremely abundant (1% of total protein) and is composed mainly of alpha subunits. It is a major component of synapses and is concentrated in a cytoskeletal structure called the postsynaptic density. When activated by a brief rise in calcium concentration, the kinase phosphorylates itself and then remains active to phosphorylate other proteins even after the calcium concentration falls. We will test the hypothesis that this is a mechanism by which long-lasting changes in synaptic function are generated following brief bursts of synaptic activity. We will determine the structure of the autophosphorylation sites by recombinant DNA and biochemical methods, then characterize the brain phosphatases responsible for dephosphorylation of each of these sites. We will raise antibodies that specifically recognize the autophosphorylated sites on the kinase and others that recognize the phosphorylated form of kinase substrates. We will use these to study, with high spatial and temporal resolution, the physiological circumstances under which the kinase is activated and specific substrates become phosphorylated. We will continue a study of the association of the kinase with the cytoskeleton by biochemical and recombinant DNA techniques. Our goal in the next few years is to clarify the possible regulatory functions of this calcium-dependent protein kinase system. Our long-term goal is to correlate information about this pathway with similar information about other calcium regulated pathways in order to understand the concerted responses to changing calcium levels in CNS neurons.

我们正在研究突触的分子结构和功能， 中枢神经系统中神经元之间的连接。 这些连接调节的紊乱是一种 一些神经系统疾病的病理学的重要组成部分， 精神疾病，包括癫痫，阿尔茨海默病， 精神分裂症和抑郁症 许多神经递质和 神经激素通过改变细胞内 钙离子水平。 我们正在研究 这些钙水平的波动改变了突触功能。 我们 将重点研究突触调节途径， 作为其中心元素，丰富的脑特异性钙， 钙调素依赖性蛋白激酶。 这种激酶是一种 两个不同但同源的催化亚基的寡聚体， α和β 在前脑，包括海马体， 在皮质和纹状体中，激酶非常丰富（1%的 总蛋白），主要由α亚基组成。 这是一个 突触的主要组成部分，并集中在一个 称为突触后密度的细胞骨架结构。 当 通过钙离子浓度的短暂升高激活， 磷酸化自身，然后保持活性磷酸化 其他蛋白质，即使在钙浓度福尔斯下降后。 我们将 测试这是一种机制，通过这种机制， 突触功能的变化是在短暂的爆发之后产生的 突触活动。 我们将确定 通过重组DNA和生物化学方法检测自身磷酸化位点 方法，然后表征脑磷酸酶负责 每个位点的去磷酸化。 我们会产生抗体 特异性识别细胞膜上的自磷酸化位点， 激酶和其他识别磷酸化形式的 激酶底物 我们将利用这些来研究，具有很高的空间 和时间分辨率， 其中激酶被激活并且特定底物变成 磷酸化 我们将继续研究 激酶与细胞骨架的生化和重组 DNA技术。 我们在未来几年的目标是澄清 这种钙依赖蛋白可能的调节功能 激酶系统 我们的长期目标是将信息 与其他钙离子类似的信息 调节途径，以了解协调的 对中枢神经系统神经元中钙水平变化的反应。