The Role of SOD Misfolding/Aggregation in Familial ALS

描述(由申请人提供)：本申请的长期目标 是为了了解人类铜基因90多个错义突变的机制 /锌超氧化物歧化酶引起家族性肌萎缩侧索硬化症 (FALS)，一种致命的运动神经系统退行性疾病。SOD1保护 细胞通过催化脱氢酶的歧化来抵抗自由基损伤 超氧自由基转化为过氧化氢和分子氧。确实是 最初认为SOD1活性的下降是导致 与sod相关的犯规；现在有压倒性的证据表明，在未知的情况下 突变型SOD1的病理功能导致了该病。而在这个时代 发病时间(约47年)随突变、病程变化不大。 发病后通常是突变依赖的，从1到20年不等。建议数 研究将检验SOD1病理功能的假设 突变体与一种异常的SOD1构象密切相关 到聚合。铜、锌的稳定性、变性机理 亲和力与全锌、载脂蛋白、缺锌的自由基产生能力 野生型和筛选的SOD1突变体的缺铜状态将是 调查过了。金属含量和自由基生活性 还将对聚合SOD1进行研究。荧光、UV/Vis和圆形 二色性光谱将被用来监测SOD1的构象变化。 SOD1聚集体的稳定性、形态和缔合率将是 通过各种技术进行研究，包括聚丙烯酰胺凝胶电泳法、UV /Vis光谱和电子显微镜。该计划的主要目的之一 拟议的研究是为了确定生物化学/生物物理效应 PALS相关的SOD1突变，并建立它们之间的相关性 FALS的影响和严重性。

DESCRIPTION (provided by the applicant): The long-term goal of this application is to understand the mechanism by which over 90 missense mutations in human Cu / Zn superoxide dismutase (SOD1) cause familial amyotrophic lateral sclerosis (FALS), a fatal degenerative disease of the motor neuron system. SOD1 protects the cell against free radical damage by catalyzing the dismutation of superoxide radicals into hydrogen peroxide and molecular oxygen. It was originally believed that a decrease in SOD1 activity was the cause of SOD-related FALS; there is now overwhelming evidence that a gain in an unknown pathological function of mutant SOD1 causes the disease. While the age of disease onset (about47 years) varies little with mutation, disease duration after onset is often mutant-dependent, ranging from 1 to 20 years. The proposed research will test the hypothesis that the pathological function of SOD1 mutants is intimately related to an abnormal SOD1 conformation that is prone to aggregation. The stability, denaturation mechanism, copper and zinc affinity, and the radical-generating ability of the holo, apo, zinc-deficient and copper-deficient states of wild type and selected SOD1 mutants will be investigated. The metal content and the radical-generating activity of aggregated SOD1 will also be studied. Fluorescence, UV / Vis, and circular dichroism spectroscopy will be used to monitor conformational changes in SOD1. The stability, morphology, and association rate of SOD1 aggregates will be studied by various techniques, including polyacrylamide gel electrophoresis, UV / Vis spectroscopy, and electron microscopy. One of the main purposes of the proposed research is to determine the biochemical/biophysical effects of PALS-related SOD1 mutations, and to establish a correlation between these effects and the severity of FALS.