Elucidation of the Biochemical Mechanism and In Vivo Functions of Spastin

Spastin的生化机制和体内功能的阐明

• 批准号: 7223823
• 负责人: Antonina Roll-Mecak
• 金额: $ 8.98万
• 依托单位: UNIVERSITY OF CALIFORNIA, SAN FRANCISCO
• 依托单位国家: 美国
• 财政年份: 2006
• 资助国家: 美国
• 起止时间: 2006-12-01 至 2008-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7223823
• 关键词: ATP Hydrolysis; ATP phosphohydrolase; Address; Affect; Architecture; Award; Binding; Biochemical; Biochemistry; Biological Assay; Biomechanics; Biophysics; Cell physiology; Cells; Cellular Structures; Cellular biology; Chemicals; Collaborations; Complex; Coupling; Cryoelectron Microscopy; Cues; Cytoskeleton; Data; Development; Disease; Electron Microscopy; Enzymes; Etiology; Eukaryotic Cell; Family; Fluorescence; Functional disorder; Genes; Goals; Hereditary Spastic Paraplegia; In Vitro; Institution; Intracellular Transport; Kinetics; Life; Light; Link; Measurement; Microtubules; Modeling; Molecular; Motor; Mutagenesis; Mutate; Mutation; Nerve Degeneration; Nervous system structure; Neurodegenerative Disorders; Neurons; Patients; Phenotype; Property; Proteins; Reaction; Regulation; Research; Research Personnel; Resolution; Series; Skeleton; Structure; Substrate Cycling; System; Techniques; Training; United States National Institutes of Health; Work; Workplace; X-Ray Crystallography; analog; axonopathy; base; concept; in vitro Assay; in vivo; mutant; neuronal survival; particle; programs; reconstruction; response; single molecule; spastin; structural biology; therapy development

DESCRIPTION (provided by applicant): The objective of this award is to complete my training and to establish a highly interdisciplinary lab at an academic institution in the U.S., focused on the study of the mechanism of cytoskeletal regulators. My lab will take a comprehensive, multifaceted approach by linking atomic resolution structural information with single molecule dynamics in vitro and in the larger context of the cell. The research focus of this award is to dissect the biochemical and cellular mechanisms of spastin. Mutations in the spastin gene are the leading cause of hereditary spastic paraplegias, a group of poorly understood neurodegenerative disorders characterized by axonopathy. I have discovered that spastin severs microtubules (Roll-Mecak and Vale, 2005). Disease mutations impair severing, linking spastin's remodeling of the microtubule cytoskeleton to neurodegeneration. The research plan outlined here is highly interdisciplinary, integrating techniques and concepts from structural biology, biophysics and cell biology to answer three fundamental questions about spastin function: 1) What is spastin's atomic structure throughout its ATPase cycle and how does it bind and break the microtubule; 2) How does it use the energy of ATP hydrolysis to disassemble the microtubule; and 3) How does spastin affect microtubule architecture and dynamics in the living cell and what are the cellular consequences incurred when spastin is depleted or mutated ? Despite its importance for neuronal survival very little is known about spastin. A detailed understanding of its mechanism and basic cell biology are critical to understanding the causes of the disease and this proposal sets the fundamental mechanistic groundwork that is needed to develop therapies further down the road. Perturbation of microtubule dynamics and architecture has emerged as a common theme in a variety of neurodegenerative diseases and an understanding of spastin's effects will have implications for the etiologies of all these disorders. Lay Summary: The goal of this research is to understand the function of an enzyme, spastin, that is defective in the majority of patients with hereditary spastic paraplegias, a group of poorly understood neurodegenerative disorders. I discovered that spastin breaks a central component of the cell's skeleton. This research will provide the basic groundwork needed for the development of treatments for this group of disorders and also increase our understanding of other neurodegenerative disorders.

描述（由申请人提供）： 这个奖项的目的是完成我的培训，并在美国的一个学术机构建立一个高度跨学科的实验室，重点研究细胞骨架调节剂的作用机制。我的实验室将采取一种全面的，多方面的方法，通过将原子分辨率结构信息与体外和细胞更大背景下的单分子动力学联系起来。该奖项的研究重点是解剖痉挛的生化和细胞机制。痉挛素基因的突变是遗传性痉挛性截瘫的主要原因，这是一组以轴突病变为特征的神经退行性疾病，人们对其了解甚少。我已经发现，spastin切断微管（Roll-Mecak and Vale，2005）。疾病突变损害切断，连接痉挛的微管细胞骨架的重塑神经变性。本研究计划是高度跨学科的，整合了结构生物学、生物物理学和细胞生物学的技术和概念，以回答关于spastin功能的三个基本问题：1）spastin在整个ATP酶循环中的原子结构是什么，它如何结合和破坏微管; 2）它如何利用ATP水解的能量来分解微管;以及3）痉挛蛋白如何影响活细胞中微管的结构和动力学，当痉挛蛋白耗尽或突变时，细胞会产生什么后果？尽管它对神经元存活的重要性，但人们对痉挛素知之甚少。详细了解其机制和基本细胞生物学对于了解疾病的原因至关重要，该提案为进一步开发治疗方法奠定了基本的机制基础。微管动力学和结构的扰动已成为各种神经退行性疾病的共同主题，对痉挛素作用的理解将对所有这些疾病的病因产生影响。简单总结：这项研究的目的是了解一种酶，痉挛素，这是有缺陷的遗传性痉挛性截瘫，一组知之甚少的神经退行性疾病的大多数患者的功能。我发现痉挛会破坏细胞骨架的一个核心组成部分。这项研究将为开发这组疾病的治疗方法提供基础，并增加我们对其他神经退行性疾病的理解。