BRAIN CALMODULIN-DEPENDENT PROTEIN KINASE

脑钙调蛋白依赖性蛋白激酶

• 批准号: 3397706
• 负责人: MARY B KENNEDY
• 金额: $ 16.07万
• 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 1987-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3397706
• 关键词: brain metabolism; chromatography; electron microscopy; electrophoresis; genetic manipulation; histochemistry /cytochemistry; immunochemistry; monoclonal antibody; neurotransmitters; phosphorylation; protein kinase; radioimmunoassay; synapses; synaptic vesicles

We are interested in 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, Huntington's chorea, 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 our studies primarily on the molecular characterization and cellular localization of a synaptic regulatory pathway that has as its central element and abundant, brain-specific calcium and calmodulin-dependent protein kinase. This enzyme is composed of two types of subunits that share certain chemical characteristics, but appear to be present in different ratios in different regions of the brain. We will compare the structure and function of the two subunits by biochemical and recombinant DNA methods. We will also compare the distributions of both subunits in different brain nuclei, quantitatively by radiommunoassay, as well as by immunohistochemistry. We will determine whether neurons containing the kinase are associated with particular transmitter systems or specific anatomical pathways. Biochemical experiments suggest that the kinase is a component of certain brain postsynaptic densities and may also be associated with synaptic vesicles and microtubules. We will determine the subcellular locations of both kinase subunits in different brain regions by biochemical and immunochemical methods and by electron microscopic immunohistochemistry. Focusing on the hippocampus, where kinase activity is most highly concentrated, we will examine the proteins that are phosphorylated by the kinase in brain slices and in homogenates. Our long term goal is two-fold. First, we want to understand the functions of this calcium-mediated regulatory pathway. Second, we want to correlate information about it with similar information about other calcium-regulated pathways (e.g. calcium-dependent adenylate cyclase, phosphodiesterase, phosphatases, etc.), in order to understand the concerted responses to changing calcium levels in CNS neurons.

我们感兴趣的是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。 混乱， 这些连接的调节是病理学的重要组成部分 几种神经和精神疾病包括癫痫，老年痴呆症 疾病、亨廷顿舞蹈症、精神分裂症和抑郁症。 许多 神经递质和神经激素调节突触功能， 细胞内钙离子水平。 我们正在研究的机制， 这些钙水平的波动会改变突触功能。 我们 我们的研究将主要集中在分子特征上， 突触调节通路的细胞定位， 中心元素和丰富的，大脑特有的钙， 钙调素依赖性蛋白激酶。 这种酶由两种类型组成 这些亚基具有某些共同的化学特征， 在大脑的不同区域有不同的比例 我们将 通过生物化学方法比较两种亚基的结构和功能， 重组DNA方法。 我们还将比较两者的分布 亚基在不同的脑核，定量放射免疫分析， 以及免疫组织化学。 我们将确定神经元 含有激酶的蛋白质与特定的递质系统有关， 特定的解剖路径 生化实验表明， 激酶是某些脑突触后密度组分，也可 与突触囊泡和微管有关。 我们将确定 两种激酶亚基在不同脑组织中的亚细胞定位 区域的生物化学和免疫化学方法和电子 显微免疫组化。 聚焦于海马体， 激酶活性是最高度集中的，我们将检查蛋白质 在脑切片和匀浆中被激酶磷酸化。 我们的长期目标是双重的。 首先，我们要了解 这一钙离子介导的调节途径。 第二，我们希望将 关于它的信息与关于其他钙调节的类似信息 途径（例如钙依赖性腺苷酸环化酶，磷酸二酯酶， 磷酸酶等），为了理解人们的一致反应， 改变中枢神经系统神经元中的钙水平。