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中90多个错义突变的机制， 锌超氧化物歧化酶（SOD 1）引起家族性肌萎缩侧索硬化症 （FALS），一种致命的运动神经元系统退行性疾病。SOD 1保护 细胞通过催化 超氧自由基转化为过氧化氢和分子氧。这是 最初认为，SOD 1活性的下降是导致 SOD相关的FALS;现在有压倒性的证据表明，在一个未知的增益 突变SOD 1的病理功能导致疾病。虽然年龄 发病时间（约47年）随突变、病程变化不大 发病后往往是依赖于肌肉，从1至20年不等。拟议 研究将检验SOD 1的病理功能 突变体与一种异常的SOD 1构象密切相关， 到聚合。稳定性、变性机理、铜和锌 亲和力，和自由基产生能力的全息，载脂蛋白，缺锌 和铜缺乏状态的野生型和选择的SOD 1突变体将被 研究了金属含量和自由基生成活性 还将研究聚集的SOD 1。荧光、UV /维斯和圆形 二色性光谱将用于监测SOD 1的构象变化。 SOD 1聚集体的稳定性、形态和缔合速率将是 研究了各种技术，包括聚丙烯酰胺凝胶电泳，紫外 /维斯光谱和电子显微镜。的主要目的之一， 拟议的研究是确定生物化学/生物物理效应 PALS相关的SOD 1突变，并建立这些突变之间的相关性。 的影响和严重程度。

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.