X-ray Studies of SOD in Amyotrophic Lateral Sclerosis

肌萎缩侧索硬化症中 SOD 的 X 射线研究

基本信息

批准号：
7800950
负责人：
Peter JOHN HART
金额：
$ 31.58万
依托单位：
UNIVERSITY OF TEXAS HLTH SCIENCE CENTER
依托单位国家：
美国
项目类别：
财政年份：
2000
资助国家：
美国
起止时间：
2000-05-05 至 2013-01-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7800950
关键词：
3-Dimensional Alzheimer&apos s Disease Amino Acid Substitution Amyloid Amyotrophic Lateral Sclerosis Animals Applications Grants Binding Binding Sites Biological Assay Brain Categories Cell Aggregation Cells Cellular Membrane Cessation of life Collaborations Congo Red Copper DNA Sequence Rearrangement Degenerative Disorder Degradation Pathway Digestion Disease Disulfides Elements Embryo Engineering Environment Enzymes Erythrocytes Familial Amyotrophic Lateral Sclerosis Fiber Filament Florida Future Goals Higher Order Chromatin Structure Human Hydrogen Peroxide Hydrophobicity In Vitro Indium Inherited Institutes Ions Laboratories Lesion Life Link Location Mammalian Cell Mediating Metalloproteins Metals Methods Mitochondria Molecular Molecular Conformation Molecular Probes Molecular Weight Motor Neuron Disease Motor Neurons Mutation Neurodegenerative Disorders Neurons Nucleosome Core Particle Oxygen Paralysed Parkinson Disease Pathogenicity Pathway interactions Patients Play Principal Investigator Process Progress Reports Property Proteins Research Personnel Research Project Grants Roentgen Rays Role SOD1 gene Scaffolding Protein Series Site Solutions Spatial Distribution Spinal Spinal Cord Structure Study Section Superoxides Surface Surface Plasmon Resonance System Testing Therapeutic Agents Tissues Toxic effect Transgenic Mice Tube Ubiquitin United States National Institutes of Health Universities Variant X ray diffraction analysis X-Ray Diffraction Yeasts Zinc absorption analytical ultracentrifugation base copper zinc superoxide dismutase cytotoxic design dimer disulfide bond falls gain of function in vivo inhibitor/antagonist interest kidney cell light scattering member monomer motor neuron degeneration mouse model multicatalytic endopeptidase complex mutant neurotoxic novel professor programs protein aggregate protein degradation protein misfolding protein oligomer research study three dimensional structure tool

项目摘要

DESCRIPTION (provided by applicant): Human copper-zinc superoxide dismutase (SOD1) is a 32 kDa homodimeric metalloprotein that catalyzes the conversion of superoxide radical into molecular oxygen and hydrogen peroxide. The enzyme is particularly abundant in red blood cells and spinal tissue. Approximately 114 different single site mutations in human SOD1 have been linked to an inherited form of amyotrophic lateral sclerosis (ALS, Lou Gehrig's disease, motor neuron disease). Both the inherited (FALS) and sporadic (SALS) forms of the disease are characterized by progressive paralysis resulting from motor neuron degeneration and death. It is now established that SOD1-linked FALS results from the gain of a cytotoxic property and not a loss of enzymatic function. Evidence is accumulating that the 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 six different metal-deficient 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 continuing project are designed to probe pathogenic SOD1 structure and to help answer the following questions: 1) What are the structural consequences of the FALS mutations and how do these amino acid substitutions render the molecule toxic? 2) Does the loss of metal ions play a role in FALS SOD1 pathogenicity? 3) How does the presence or absence of the intrasubunit disulfide bond influence the structural and biophysical properties of pathogenic SOD1? 4) Is the mode of self-association we observe in X-ray studies the basis for how the pathogenic SOD1 proteins aggregate in living cells? 5) What are the structural elements of SOD1 proteins that are recognized by the 20 S proteasome so that they may be degraded? 6) Could soluble oligomers (protofibrils) and/or insoluble amyloids of pathogenic SOD1 act as proteasomal inhibitors? Answers to questions such as these are required for a molecular understanding of SOD1 linked FALS and for the design of therapeutic agents aimed inhibiting the aggregation process.

描述（由申请人提供）：人铜锌超氧化物歧化酶（SOD1）是一种 32 kDa 同型二聚体金属蛋白，可催化超氧自由基转化为分子氧和过氧化氢。这种酶在红细胞和脊髓组织中特别丰富。人类 SOD1 中大约 114 个不同的单位点突变与遗传性肌萎缩侧索硬化症（ALS、卢伽雷氏病、运动神经元病）有关。该疾病的遗传性（FALS）和散发性（SALS）形式的特征都是运动神经元变性和死亡导致的进行性瘫痪。现在已经确定，SOD1 相关的 FALS 是由于细胞毒性特性的获得而不是酶功能的丧失所致。越来越多的证据表明，SOD1 突变蛋白的毒性来自于其组装成更高阶结构（可溶性低聚物和不溶性聚集体）的能力，从而以某种方式干扰神经元细胞机制。使用成熟的单晶 X 射线衍射工具，我们最近观察到六种不同的金属缺陷型 FALS 突变体 SOD1 蛋白可以形成“淀粉样蛋白样”纤维，这在某种程度上让人想起其他神经退行性疾病中看到的纤维类型，例如阿尔茨海默病和帕金森病。致病性 SOD1 蛋白能够自结合，但正常的、未突变的 SOD1 蛋白则不能。这个持续项目中概述的实验旨在探测致病性 SOD1 结构并帮助回答以下问题：1) FALS 突变的结构后果是什么以及这些氨基酸取代如何使分子有毒？ 2) 金属离子的丢失是否对FALS SOD1的致病性起作用？ 3) 亚基内二硫键的存在与否如何影响致病性SOD1的结构和生物物理特性？ 4) 我们在 X 射线研究中观察到的自关联模式是致病性 SOD1 蛋白如何在活细胞中聚集的基础吗？ 5) SOD1 蛋白的哪些结构元件可以被 20 S 蛋白酶体识别从而被降解？ 6) 致病性 SOD1 的可溶性低聚物（原纤维）和/或不溶性淀粉样蛋白能否充当蛋白酶体抑制剂？回答诸如此类的问题是从分子角度理解 SOD1 相关 FALS 以及设计旨在抑制聚集过程的治疗药物所必需的。

项目成果

期刊论文数量（21）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

Structures of the G85R variant of SOD1 in familial amyotrophic lateral sclerosis.

家族性肌萎缩侧索硬化症中 SOD1 G85R 变体的结构。

DOI：
发表时间：
2008-06-06
期刊：
影响因子：
0
作者：
Cao, Xiaohang;Antonyuk, Svetlana V;Seetharaman, Sai V;Whitson, Lisa J;Taylor, Alexander B;Holloway, Stephen P;Strange, Richard W;Doucette, Peter A;Valentine, Joan Selverstone;Tiwari, Ashutosh;Hayward, Lawrence J;Padua, Shelby;Cohlberg, Jeffrey
通讯作者：
Cohlberg, Jeffrey

Immature copper-zinc superoxide dismutase and familial amyotrophic lateral sclerosis.

未成熟的铜锌超氧化物歧化酶和家族性肌萎缩侧索硬化症。

DOI：
发表时间：
2009-10
期刊：
影响因子：
0
作者：
Seetharaman, Sai V;Prudencio, Mercedes;Karch, Celeste;Holloway, Stephen P;Borchelt, David R;Hart, P John
通讯作者：
Hart, P John

Cobalt(2+) binding to human and tomato copper chaperone for superoxide dismutase: implications for the metal ion transfer mechanism.

钴 (2 ) 与人和番茄铜伴侣结合形成超氧化物歧化酶：对金属离子转移机制的影响。

DOI：
发表时间：
2000-05-09
期刊：
影响因子：
2.9
作者：
Zhu, H;Shipp, E;Sanchez, R J;Liba, A;Stine, J E;Hart, P J;Gralla, E B;Nersissian, A M;Valentine, J S
通讯作者：
Valentine, J S

Structural consequences of the familial amyotrophic lateral sclerosis SOD1 mutant His46Arg.

家族性肌萎缩侧索硬化症 SOD1 突变体 His46Arg 的结构后果。

DOI：
发表时间：
2005-05
期刊：
影响因子：
0
作者：
Antonyuk, Svetlana;Elam, Jennifer Stine;Hough, Michael A;Strange, Richard W;Doucette, Peter A;Rodriguez, Jorge A;Hayward, Lawrence J;Valentine, Joan Selverstone;Hart, P John;Hasnain, S Samar
通讯作者：
Hasnain, S Samar

Proteasomal degradation of mutant superoxide dismutases linked to amyotrophic lateral sclerosis.

突变型超氧化物歧化酶的蛋白酶体降解与肌萎缩侧索硬化症有关。