Structural and Biophysical Properties of Oligermeric SOD1

寡聚 SOD1 的结构和生物物理特性

基本信息

批准号：
7551910
负责人：
P J HART
金额：
$ 33.94万
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：
资助国家：
美国
起止时间：
至
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/7551910
关键词：
3-Dimensional Address Amino Acid Substitution Amyloid Amyotrophic Lateral Sclerosis Animals Anus Apoproteins Binding Binding Sites Biochemistry Biological Assay C-terminal Categories Cell Aggregation Cell model Chemistry Class Collaborations Congo Red Consensus Copper Cultured Cells DNA Sequence Rearrangement Degenerative Disorder Degradation Pathway Disease Dissociation Electrostatics Elements Embryo Engineering Enzymes Familial Amyotrophic Lateral Sclerosis Filament Future Goals Human Hydrogen Peroxide Impairment In Vitro Indium Inherited Investigation Ions Laboratories Lead Lesion Light Link Location Maps Mass Spectrum Analysis Mediating Metal Binding Site Metals Methodology Methods Modeling Modification Molecular Molecular Conformation Molecular Probes Monitor Motor Neurons Mus Mutation Nature Neurodegenerative Disorders Neurons Numbers Patients Peroxidase Peroxidases Phosphines Principal Investigator Process Production Property Proteins Published Comment Range Reaction Readability Research Research Design Research Personnel Research Project Grants Roentgen Rays Role SOD1 gene Scaffolding Protein Series Site Solubility Solutions Spatial Distribution Spinal Cord Stress Structure Study Section Superoxide Dismutase Superoxides Surface Plasmon Resonance Technical Expertise Techniques Testing Therapeutic Agents Thinking Time Tissues Toxic effect Transgenic Animals Transgenic Mice Transgenic Organisms Variant Work Writing X ray diffraction analysis X-Ray Diffraction analytical ultracentrifugation base copper zinc superoxide dismutase crosslink deletion analysis design dimer disulfide bond editorial falls gain of function improved in vivo innovation insight interest kidney cell lead ion light scattering member monomer motor neuron degeneration mouse model mutant neurotoxicity novel oxidation phosphine prevent programs protein aggregation protein degradation protein misfolding protein oligomer reaction rate research study response structural biology three dimensional structure tissue culture tool

项目摘要

Since the link between mutations in copper-zinc superoxide dismutase (SO01) and familial ALS (FALS) was first described approximately 10 years ago, laboratories worldwide have sought to understand how the mutations render the SOD1 protein toxic to motor neurons. Evidence is accumulating that this toxic property comes from the ability of the mutant SOD1 proteins to assemble into higher order structures (soluble oligomers and insoluble aggregates) that somehow interfere with the neuronal cellular machinery. Using the well established tools of single crystal X-ray diffraction, we recently observed that five different FALS mutant SOD1 proteins can form "amyloid-like" fibers that are somewhat reminiscent of the types of fibers seen in other neurodegenerative disorders such as Alzheimer's and Parkinson's diseases. The pathogenic SOD1 proteins are able to self-associate but the normal, unmutated SOD1 proteins cannot. The experiments outlined in this project are designed to probe these abnormal pathogenic SOD1-SOD1 interactions and to help answer the following questions: 1) Is the mode of self-association we observe in X-ray studies the basis for how the pathogenic SO01 proteins aggregate in living cells? 2) What structural elements are determinants of toxicity in pathogenic SOD1? 3) Does oxidative modification of pathogenic SOD1 play a role in aggregation? 4) Could a dissociated (monomeric) form of pathogenic SOD1 be responsible for aggregation? Answers to questions such as these are required for the design of therapeutic agents aimed inhibiting the aggregation process.

由于铜锌超氧化物歧化酶 (SO01) 突变与家族性 ALS (FALS) 之间的联系大约 10 年前首次被描述，世界各地的实验室一直在试图了解突变使 SOD1 蛋白对运动神经元有毒。越来越多的证据表明这种有毒物质来自突变体 SOD1 蛋白组装成更高阶结构（可溶性寡聚体）的能力和不溶性聚集体）以某种方式干扰神经元细胞机制。使用井建立单晶X射线衍射工具后，我们最近观察到五种不同的FALS突变体SOD1 蛋白质可以形成“淀粉样蛋白样”纤维，这在某种程度上让人想起其他生物中看到的纤维类型神经退行性疾病，例如阿尔茨海默病和帕金森病。致病性 SOD1 蛋白能够自我结合，但正常的、未突变的 SOD1 蛋白却不能。本文概述的实验该项目旨在探测这些异常致病性 SOD1-SOD1 相互作用并帮助回答以下问题：1）我们在 X 射线研究中观察到的自联想模式是否是致病性 SO01 蛋白在活细胞中聚集？ 2) 哪些结构元素是毒性的决定因素致病性SOD1？ 3）致病性SOD1的氧化修饰是否在聚集中发挥作用？ 4）致病性 SOD1 的解离（单体）形式是否会导致聚集？答复设计旨在抑制聚集的治疗剂需要解决诸如此类的问题过程。