The Role of SOD Misfolding/Aggregation in Familial ALS

SOD 错误折叠/聚集在家族性 ALS 中的作用

基本信息

批准号：
6853605
负责人：
WILFREDO COLON
金额：
$ 24.04万
依托单位：
RENSSELAER POLYTECHNIC INSTITUTE
依托单位国家：
美国
项目类别：
财政年份：
2002
资助国家：
美国
起止时间：
2002-01-15 至 2006-12-31
项目状态：
已结题

项目摘要

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.

描述（申请人提供）：本申请的长期目标是要了解人类Cu中90多个错义突变的机制 / Zn超氧化物歧化酶（SOD1）引起家族性肌萎缩性侧面硬化症（伪造），一种运动神经元系统的致命退化性疾病。 SOD1保护通过催化拆除的损害来抵抗自由基损害的细胞超氧化物自由基进入过氧化氢和分子氧。是最初认为SOD1活性的减少是原因与SOD相关的伪装；现在有很多证据表明未知的收益突变SOD1的病理功能会导致该疾病。而年龄疾病发作（约47年）随着突变，疾病持续时间而异发作后通常是突变体依赖性的，范围为1至20年。提议研究将检验SOD1的病理功能的假设突变体与俯卧的异常SOD1构象密切相关聚集。稳定性，变性机制，铜和锌亲和力，以及holo，apo，锌缺陷的自由基生成能力野生型和选定的SOD1突变体的缺陷状态和缺乏铜状态将是调查。金属含量和自由基生成活性聚集的SOD1也将进行研究。荧光，紫外线 / vis和圆形二分色谱法将用于监测SOD1中的构象变化。 SOD1聚集体的稳定性，形态和关联率将是通过各种技术研究，包括聚丙烯酰胺凝胶电泳，UV / VIS光谱和电子显微镜。主要目的之一拟议的研究是确定与PALS相关的SOD1突变，并在这些突变之间建立相关性影响和伪造的严重程度。