Project 1

项目1

基本信息

批准号：
8249450
负责人：
JOAN Selverstone VALENTINE
金额：
$ 22.6万
依托单位：
UNIVERSITY OF CALIFORNIA LOS ANGELES
依托单位国家：
美国
项目类别：
财政年份：
2005
资助国家：
美国
起止时间：
2005-08-11 至 2015-04-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8249450
关键词：
Address Affect Age Amyloid Fibrils Amyotrophic Lateral Sclerosis Animal Disease Models Animal Model Animals Award Binding Biochemical Biological Assay Biological Models Blood Brain Cell Culture Techniques Cells Cessation of life Characteristics Collaborations Copper Dependence Deuterium Diagnostic radiologic examination Disease Disease Progression Disulfides Elements Ensure Equilibrium Etiology Exposure to Familial Amyotrophic Lateral Sclerosis Fluorescence Microscopy Fourier Transform Future Gene Mutation Goals Homeostasis Human Hydrogen Hydrophobic Surfaces Hydroxyl Radical Image In Vitro Ions Knowledge Label Laboratories Lead Light Link Liver Measures Metals Methods Modeling Molecular Molecular Structure Molecular Weight Monitor Motor Neurons Mus Mutation National Institute of Diabetes and Digestive and Kidney Diseases Nerve Degeneration Neurodegenerative Disorders Oxidation-Reduction Patients Peptide Hydrolases Predisposition Preparation Process Property Proteins Protocols documentation Reaction Reagent Recruitment Activity Relative (related person)Reproducibility Roentgen Rays Role Seeds Site Solubility Source Spinal Spinal Cord Spleen Staging Structure Study models Surveys Symptoms Synchrotrons Techniques Testing Tissues Toxic effect Toxicity Tests Transgenic Mice Transgenic Organisms Translational Research Work Zinc animal tissue base beta pleated sheet copper zinc superoxide dismutase gray matter human embryonic stem cell human tissue monomer mouse model mutant physical property programs protein aggregate research study small molecule solid state nuclear magnetic resonance tissue/cell culture tool white matter

项目摘要

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.

肌萎缩性外侧硬化症（ALS）是一种进行性致命的神经退行性疾病，其为特征运动神经元的选择性死亡。虽然最常见的ALS形式是零星的，但没有已知原因，但由基因突变引起的一部分病例是家族性的，其中那些是由突变引起的蛋白质铜锌超氧化物歧化酶（SODL）代表了最广泛研究的ALS模型。这脊髓中富含SODL的原纤维夹杂物的形成是Sodi连接家族的重要特征该疾病的人类患者和动物模型中的ALS。在动物模型中，包含在高分子重量的sodl的高分子寡聚形式的形成，甚至在症状，表明SODL的低聚和聚集是疾病病因。了解多聚体S0D1如何促进运动神经元死亡是总体计划项目的目标。在这个项目中，我们将解决SODL多聚化的生物物理方面。具体而言，目标包括（1）检查在体外或孤立的多聚体SODL的结构（2）从人类和动物组织来源应用标记的SODL的定义多聚体制剂培养的运动神经元可以研究它们是否有毒（与项目2合作），（3）检查 S0D1多聚化的机理成原纤维，以了解如何破坏SODL的结构因素有助于这一过程，（4）阐明由家族性ALS引起的突变在调节中的作用这些过程的速率。我们的研究将广泛使用我们在先前奖项中开发的测定法将SODL转化为可溶性，低聚物种和淀粉样纤维的时期相关条件。我们还将广泛使用多种高度敏感的生物物理方法研究各种结构特性，例如折叠，金属含量和可溶性和二硫化状态从动物组织中分离出的SODL的不溶性形式。