HUMAN SPINAL CORD PROTEIN IDENTIFICATION

人类脊髓蛋白鉴定

• 批准号: 6120272
• 负责人: DON W CLEVELAND
• 金额: $ 1.77万
• 依托单位: UNIVERSITY OF CALIFORNIA, SAN FRANCISCO
• 依托单位国家: 美国
• 财政年份: 1999
• 资助国家: 美国
• 起止时间: 1999-03-01 至 2000-02-29
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6120272
• 关键词: biomedical resource; nervous system; proteins; spectrometry; structural biology

The research interests of the Cleveland group focus on mechanisms of neuronal growth and death using molecular genetics to assess how axons grow and what factors provoke selective death of motor neurons. Unlike most eukaryotic cells, an intrinsic feature of neurons is their extreme asymmetry. for example, in the human peripheral nervous system, single motor neurons extend over a meter in length. Asymmetry is achieved in two phases. The first is when a neurite extends towards its target. After stable synapse formation, a second phase initiates in which the axon grows up to ten fold in diameter. This radial growth phase, concomitant with meylination and essential for establishment of proper conduction velocity, yields an enormous increase in axonal volume and a huge cell, 99.9% of which is in the axon. We have focused part of our effort on using molecular genetics and transgenic mice to test how radial growth of axons is achieved. Already we have shown that neurofilaments, the most abundant structural element in axons, are essential for radial growth. We have now identified a set of neurofilament-associated proteins and, in collaboration with Dr. Burlingame, now wish to use modern protein analysis methods to determine the identities of these associated proteins. Once the corresponding genes are isolated we will then use transgenic and gene disruption methods in mice to determine the corresponding in vivo properties. Beyond this, essentially all human motor neuron disorders are characterized by the maldistribution of neurofilaments, a finding clearly suggesting that they may play an essential role in disease pathogenesis. This is particularly true in the most prominent motor neuron disease, amyotrophic lateral sclerosis, or ALS, which is characterized by selective death of motor neurons. By producing transgenic mice expressing mutations in neurofilaments, we have proven that such mutations can cause ALS in mice and we are now searching for the presence of similar mutations in human patients. The cause of only 1.5% of human ALS is known and this is point mutations in an enzyme superoxide dismutase. By expression of these mutations in mice, we have proven that disease arises from a toxic property of the mutant proteins and we are now looking for what that property is and what is the cascade of events leading to selective motor neuron death. By using high resolution protein analysis methods, we now seek to determine whether neurofilaments are indeed targets for covalent damage arising from ALS-linked SOD1 mutations, and if so, to identify the nature of the aberrant chemistry mediated by these mutant proteins.

克利夫兰小组的研究兴趣集中在机制 神经元生长和死亡的研究， 轴突生长以及什么因素引起运动神经元的选择性死亡。 与大多数真核细胞不同，神经元的一个内在特征是它们的 极度不对称 例如，在人类外周神经中， 系统中，单个运动神经元的长度超过一米。 不对称 分两个阶段实现。 第一种是当神经突 朝着目标前进 在稳定的突触形成之后， 轴突直径增长到十倍的起始阶段。 这 径向生长阶段，伴随着meyelets和必不可少的 建立适当的传导速度，会产生巨大的 轴突体积增加和巨大的细胞，其中99.9%在 轴突 我们把部分精力集中在利用分子遗传学上 和转基因小鼠来测试轴突的径向生长是如何实现的。 我们已经证明，最丰富的神经丝 轴突中结构元件是径向生长所必需的。 我们有 现在确定了一组神经毒性相关蛋白， 与伯林盖姆博士合作，现在希望使用现代蛋白质 分析方法，以确定这些相关的身份 proteins. 一旦相应的基因被分离出来，我们将使用 转基因和基因破坏方法，以确定 相应的体内特性。 除此之外，基本上所有人类 运动神经元疾病的特征在于 神经丝，这一发现清楚地表明，它们可能发挥作用， 在疾病发病机制中的重要作用。 尤其如此 最突出的运动神经元疾病，肌萎缩侧索硬化， 硬化症，或ALS，其特征是运动神经的选择性死亡， 神经元 通过生产转基因小鼠， 我们已经证明，这种突变可以导致ALS， 我们现在正在寻找类似突变的存在， 人类病人 只有1.5%的人类ALS的原因是已知的， 是超氧化物歧化酶的点突变 由表达式 在小鼠的这些突变中，我们已经证明疾病是由一种 突变蛋白质的毒性，我们现在正在寻找什么 这一属性是什么， 选择性运动神经元死亡 通过使用高分辨率蛋白质 分析方法，我们现在试图确定神经丝是否 事实上，ALS连接的SOD 1引起的共价损伤的靶点 突变，如果是这样，以确定异常化学的性质， 由这些突变蛋白质介导。