Nascent SOD1 in Amyotrophic Lateral Sclerosis

肌萎缩侧索硬化症中的新生 SOD1

基本信息

批准号：
8195922
负责人：
Peter JOHN HART
金额：
--
依托单位：
SOUTH TEXAS VETERANS HEALTH CARE SYSTEM
依托单位国家：
美国
项目类别：
财政年份：
2009
资助国家：
美国
起止时间：
2009-10-01 至 2014-06-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8195922
关键词：
3-Dimensional Address Adopted Affect Affinity Age Aging-Related Process Amino Acid Substitution Amino Acids Amyotrophic Lateral Sclerosis Antioxidants Behavior Binding Binding Sites Biochemistry Cessation of life Child Complex Consultations Copper Cultured Cells DNA Sequence Rearrangement Data Defect Degenerative Disorder Dimensions Disease Dissociation Disulfides Enzymes Event Exercise Failure Familial Amyotrophic Lateral Sclerosis Family Future Gel Chromatography Generations Genes Health Sciences Hospitals Human Inherited Ions Iraq Laboratories Lead Lesion Letters Light Link Measurement Mediating Metalloproteins Metals Methods Military Personnel Modeling Molecular Molecular Chaperones Molecular Conformation Motion Motor Neuron Disease Motor Neurons Mus Mutation Neurodegenerative Disorders Neurons Paralysed Parents Pathway interactions Patients Population Post-Translational Protein Processing Process Proline Proteins Relative (related person)Reporting Resolution SOD1 gene Sampling Site Solutions Spinal Cord Structure Study Section Temperature Testing Texas Therapeutic Therapeutic Agents Time Toxic effect Transgenic Mice Translating United States Universities Variant Veterans War Work X ray diffraction analysis X-Ray Crystallography X-Ray Diffraction analytical ultracentrifugation base cofactor conformer copper zinc superoxide dismutase design disease-causing mutation disulfide bond gain of function gel electrophoresis human tissue in vivo interest melting molecular mass mouse model mutant oxidation polypeptide protein aggregation protein misfolding public health relevance relating to nervous system research study sedimentation equilibrium sedimentation velocity tool

项目摘要

Mutations in human copper-zinc superoxide dismutase (SOD1) cause an inherited form of the fatal neurodegenerative disease amyotrophic lateral sclerosis (ALS, Lou Gehrig's disease, motor neuron disease). With increasing age, insoluble forms of mutant SOD1 progressively accumulate in neural tissues of human ALS patients and in spinal cords of transgenic mice expressing these polypeptides, suggesting that SOD1- linked ALS is a protein misfolding and aggregation disorder. Understanding the molecular basis for how the pathogenic mutations give rise to SOD1 folding intermediates that are prone to aggregation is therefore of keen interest. A critical step on the folding pathway occurs when the nascent SOD1 polypeptide is posttranslationally modified by the copper chaperone for SOD1 (CCS), a helper protein that inserts the catalytic copper cofactor and oxidizes the SOD1 intrasubunit disulfide bond. CCS recognizes and binds to nascent, but not mature SOD1, suggesting that the newly translated form exists in a conformation distinct from the mature form. Recent studies reveal that pathogenic SOD1 proteins coming from aggregates isolated from cultured cells and from the spinal cords of transgenic mice tend to be metal-deficient and/or lacking the disulfide bond. These observations suggest the hypothesis that the disease-causing mutations may enhance levels of SOD1 folding intermediates by hindering CCS-mediated SOD1 maturation. The experiments outlined in this project are designed to probe the structure and dynamics of nascent SOD1 and to examine its interaction with CCS. Successful completion of the Aims contained herein will address the following questions: 1) What are the structural determinants that are responsible for the recognition of nascent SOD1 by CCS? 2) Is the functional SOD1/CCS complex heterodimeric, heterotetrameric, or some other higher order oligomer? 3) What are the structural details of the nascent SOD1/CCS complex in three dimensions? 4) What factors govern the interconversion of the nascent and mature SOD1 conformations in the absence of CCS? 5) How do the ALS mutations affect the interconversion of the nascent and mature conformations? Answers to these questions are prerequisites for the design of therapeutic agents aimed inhibiting pathogenic SOD1 aggregation in ALS. 1

人类铜 - 锌超氧化物歧化酶（SOD1）的突变导致致命的遗传形式神经退行性疾病肌萎缩性侧面硬化症（ALS，Lou Gehrig病，运动神经元疾病）。随着年龄的增长，突变体SOD1的不溶性形式逐渐积聚在人类的神经组织中 ALS患者和表达这些多肽的转基因小鼠的脊髓中，表明SOD1- 链接的ALS是一种蛋白质错误折叠和聚集障碍。了解分子基础因此，致病突变引起了容易汇总的SOD1折叠中间体敏锐的兴趣。当新生的SOD1多肽为通过铜伴侣修饰的翻译后，用于SOD1（CCS），辅助蛋白插入催化铜辅因子并氧化SOD1亚基二硫键。 CCS识别并结合到新生但不是成熟的SOD1，表明新翻译的形式存在于不同的构象中成熟的形式。最近的研究表明，来自从中分离的聚集体的致病SOD1蛋白培养的细胞和转基因小鼠的脊髓往往是金属缺陷和/或缺乏的二硫键。这些观察结果表明，引起疾病的突变可能会增强的假设 SOD1折叠中间体的水平通过阻碍CCS介导的SOD1成熟。实验概述了在该项目中，旨在探测新生SOD1的结构和动力学并检查其与CCS相互作用。成功完成此处包含的目标将解决以下问题： 1）CCS识别新生SOD1的结构决定因素是什么？ 2）功能性SOD1/CCS复合物异二聚体，杂词还是其他高阶低聚物？ 3）在三个维度上，新生SOD1/CCS复合物的结构细节是什么？ 4）什么因素在没有CCS的情况下，控制新生和成熟的SOD1构象的互连？ 5）如何 ALS突变会影响新生和成熟构象的互连吗？这些答案问题是设计治疗剂的先决条件，旨在抑制致病性SOD1聚集在ALS中。 1