CYTOSKELETAL OXIDATIVE MODIFICATIONS

细胞骨架氧化修饰

• 批准号: 2396694
• 负责人: LAWRENCE M SAYRE
• 金额: $ 21.1万
• 依托单位: CASE WESTERN RESERVE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 1997
• 资助国家: 美国
• 起止时间: 1997-08-15 至 2000-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2396694
• 关键词: carbonyl group; crosslink; human tissue; mass spectrometry; neurofilament proteins; oxidation; protein structure function; tau proteins; tissue /cell culture

DESCRIPTION (Adapted from applicant's abstract): Despite an extensive research directed at characterizing the cytoskeletal protein defects associated with aging and age-related neurodegeneration, the nature of the modifications responsible for protein deposition, insolubility, and proteolytic resistance remain elusive. Of particular relevance are neurofibrillary tangles in Alzheimer's disease, spheroids in amyotrophic lateral sclerosis, and Lewy bodies in Parkinson's disease. Although one or more factors appear to induce a transition between monomeric and non-covalent-associated forms of the constituent proteins, mainly neurofilaments (NF) or tau, the applicant postulates, and has preliminary evidence, that oxidative stress processes lead to an irreversible "cementing" of protein aggregates, at least in part, through covalent crosslinking. Modification involves a combination of direct oxidation of protein side chains and adduction of products of glycoxidation and lipid peroxidation that is expected to be manifested in overlapping but distinct patterns of markers for the three disease states. Thus, knowledge of individual disease variations in the structure and extent of these markers is hypothesized to help clarify the mechanisms of disease pathogenesis and provide clues for designing ameliorative agents/approaches. The proposal represents a multidisciplinary collaboration among three mid-career investigators with expertise in organic/peptide chemistry (Sayre), protein/analytical biochemistry (Anderson), and neuropathology/immunobiochemistry (Perry), with the goal of defining the structural basis of irreversible protein deposition. The aims are (i) structural elucidation of the crosslink and non-crosslink-carbonyl oxidative modifications in the protein deposits using novel mass-spectral biochemical analyses, (ii) development of antibodies to oxidation-specific NF and tau conformational epitopes and to structurally defined oxidative lesions, and (iii) correlative evaluation of spatio-temporal distribution of the oxidative-specific markers across different brain regions and their pathobiological significance.

描述（改编自申请人的摘要）：尽管有广泛的 旨在表征细胞骨架蛋白缺陷的研究 与衰老和年龄相关的神经变性有关， 导致蛋白质沉积、不溶性和 蛋白水解抗性仍然难以捉摸。 特别相关的是 阿尔茨海默病中的神经原纤维缠结，肌萎缩症中的球体 侧索硬化症和帕金森病中的路易体。 虽然一或 更多的因素似乎会诱导单体和单体之间的转变 组成蛋白的非共价结合形式，主要是 申请人假设神经丝（NF）或 tau 蛋白，并已初步确定 有证据表明，氧化应激过程会导致不可逆的 蛋白质聚集体的“胶结”，至少部分是通过共价键 交联。 改性涉及直接氧化的组合 蛋白质侧链以及糖氧化和脂质产物的加合 预期表现为重叠但不同的过氧化反应 三种疾病状态的标记模式。 因此，知识 这些标志物的结构和范围存在个体疾病差异 假设有助于阐明疾病发病机制和 为设计改善剂/方法提供线索。 提案 代表了三个职业生涯中期之间的多学科合作 具有有机/肽化学专业知识的研究人员（Sayre）， 蛋白质/分析生物化学（安德森），以及 神经病理学/免疫生物化学（Perry），目标是定义 不可逆蛋白质沉积的结构基础。 目标是 (i) 交联和非交联-羰基氧化的结构解析 使用新型质谱生化技术对蛋白质沉积物进行修饰 分析，(ii) 开发针对氧化特异性 NF 和 tau 的抗体 构象表位和结构上确定的氧化损伤，以及 (iii) 时空分布的相关评价 不同大脑区域的氧化特异性标记物及其 病理生物学意义。