Project 1

肌萎缩侧索硬化症（ALS）是一种进行性、致命性神经退行性疾病，其特征在于： 运动神经元的选择性死亡虽然最常见的ALS形式是散发性的，没有已知的原因， 由基因突变引起的一部分病例是家族性的，其中由基因突变引起的病例是家族性的。 蛋白质铜-锌超氧化物歧化酶（SOD 1）代表ALS的最广泛研究的模型。的 在脊髓中形成富含SOD 1的纤维状内含物是与SOD 1相关的家族性脊髓损伤的一个突出特征。 人类患者和这种疾病的动物模型中的ALS。在动物模型中，内含物之前是 形成高分子量的低聚形式的SOD 1，甚至出现在发病之前， 症状，这表明SOD 1的寡聚化和聚集是SOD 1的重要组成部分。 疾病病因学了解多聚体S 0 D1如何促进运动神经元死亡是至关重要的 计划项目的目标。在这个项目中，我们将解决SOD 1多聚化的生物物理方面。 具体地，目标包括（1）检查体外产生或分离的多聚体SOD 1的结构， 从人和动物组织来源，（2）将标记的SOD 1的限定的多聚体制剂应用于 培养运动神经元，研究它们是否有毒以及如何有毒（与项目2合作），（3）检查 SOD 1多聚化成纤维的机制，以了解使SOD 1不稳定的结构因子 有助于这一过程，（4）阐明家族性ALS引起的突变在调节 这些过程的速度。我们的研究将广泛使用我们在先前的裁决中开发的一种测定方法 在温和的生理条件下将SOD 1转化为可溶性低聚物和淀粉样纤维的时间 相关条件。我们还将广泛使用各种高灵敏度的生物物理方法， 研究各种结构特性，如折叠，金属含量和可溶性和 从动物组织分离的不溶性形式的SOD 1。

Amyotrophic lateral sclerosis (ALS) is a progressive, fatal neurodegenerative disease characterized by the selective death of motor neurons. While the most common form of ALS is sporadic and has no known cause, a subset of cases caused by genetic mutations are familial, of which those caused by mutations in the protein copper-zinc superoxide dismutase (SODl) represent the most extensively studied model of ALS. The formation of SODl-rich fibrillar inclusions in the spinal cord is a prominent feature of SODI-linked familial ALS in human patients and animal models of this disease. In animal models, the inclusions are preceded by the formation of high-molecular-weight oligomeric forms of SODl that appear even before the onset of symptoms, suggesting that oligomerization and aggregation of SODl is an essential component of the disease etiology. Understanding how multimeric S0D1 contributes to motor neuron death is the overarching goal of the Program Project. In this project, we will address the biophysical aspects of SODl multimerization. Specifically, the goals include (1) examining the structure of multimeric SODl generated in vitro or isolated from human and animal tissue sources, (2) applying defined multimeric preparations of tagged SODl to cultured motor neurons to study if and how they are toxic (in collaboration with project 2), (3) examining the mechanism of S0D1 multimerization into fibrils to understand how structural factors that destabilize SODl contribute to this process and, (4) elucidating the role of familial ALS-causing mutations in modulating the rate of these processes. Our studies will make extensive use of an assay we developed in the prior award period for converting SODl into soluble, oligomeric species and amyloid fibrils under mild, physiologically relevant conditions. We will also make extensive use of a variety of highly sensitive biophysical methods to study a variety of structural properties such as folding,.metal content, and disulfide status of soluble and insoluble forms of SODl isolated from animal tissues.