Molecular Mechanisms of SOD1-linked ALS (P01)

铜锌超氧化物歧化酶(SOD1)错义突变与常染色体相关- 以成人起病的家族性肌萎缩侧索硬化症(FALS)。的主要调查人员 此应用程序计划采用一种协调、协同的方法来确定这些SOD1突变如何 与肌萎缩侧索硬化症有关。J.S.瓦伦丁博士的实验室在SOD1研究方面有着长期的记录 矿物生物化学。P.J.哈特博士的实验室在3D结构方面有10多年的经验， 正常和致病SOD1的功能研究。D.R.Borchelt博士有既定的记录 利用转基因小鼠研究SOD1连锁FALS的机制。在一起，我们将使用三- 突变型SO1毒力机制剖析(S)-化学(项目1)，3- D结构和溶液的生物物理性质(项目2)，以及特定疾病的功能(项目3)。 为了支持这些项目，我们将拥有一个核心，它以以下形式提供分析支持 蛋白质组学、质谱学和电感耦合等离子体质谱金属分析。我们的总体做法是 编码针对特定化学或结构的新突变的SOD1蛋白的特征 蛋白质的特征。在彻底的体外鉴定之后，我们将在体内检测 这些新的SOD1蛋白导致运动神经元疾病。该项目的目标是：1)了解 氧化化学在运动神经元直接和间接毒性模式中的作用 了解SOD1氧化损伤对其结构的影响及解决方法 属性；3)确定SOD1中使其易于生成非本机的结构元素 同(或异)聚合物相互作用，并了解它们在运动神经元毒性中的作用； 和4)通过表征突变的超氧化物歧化酶聚集的原因和后果 在这些聚集体中发现的SOD1蛋白质，以及通过鉴定其他蛋白质 可能包含在这些结构中。通过我们的共同努力，我们将能够 结构和疾病-功能关系达到了前所未有的水平。

Missense mutations in copper-zinc superoxide dismutase (SOD1) are linked to autosomal- dominant, adult-onset, familial amyotrophic lateral sclerosis (FALS). The principal investigators of this application plan a concerted,synergistic approach to determine how these SOD1 mutations relate to ALS. Dr. J.S. Valentine's laboratory has a long track record in the study of SOD1 metallobiochemistry. Dr. P.J. Hart's laboratory has over 10 years experience in 3-D structure and function studies of normal and pathogenic SOD1. Dr. D.R. Borchelt has an established track record in using transgenic mice to study mechanisms of SOD1-linked FALS. Together, we will use a three- pronged approach to dissecting the toxic mechanism(s) of mutant SOD1 - chemistry (Project 1),3- D structure and solution biophysical properties (Project 2), and disease-specific function (Project 3). To support these projects, we will have a Core that provides analytical support in the form of proteomics, mass spectrometry, and ICP-MS metal analysis. Our general approach is to characterize SOD1 proteins encoding novel mutations that target a particular chemical or structural feature of the protein. After thorough in vitro characterization, we will examine, in vivo, the ability of these novel SOD1 proteins to cause motor neuron disease. The Project goals are to: 1) understand the role of oxidative chemistry in both direct and indirect modes of toxicity to motor neurons; 2) understand the consequences of oxidative damage to SOD1 on its structure and its solution properties; 3) determine the structural elements in SOD1 that predispose it to produce non-native homo- (or hetero-) polymeric interactions, and to understand their role in toxicity to motor neurons; and 4) understand the causes and consequences of mutant SOD aggregation by characterizing the SOD1 protein that is found within these aggregates as well as by characterizing other proteins that may be contained within these structures. Through our combined efforts we will be able to define structure and disease-function relationships at an unprecedented level.