CYTOSKELETAL OXIDATIVE MODIFICATIONS

描述(改编自申请人的摘要)：尽管有广泛的 针对细胞骨架蛋白缺陷的研究 与衰老和年龄相关的神经退行性变有关， 修饰导致蛋白质沉积、不溶性和 对蛋白质降解的抵抗力仍然难以捉摸。特别相关的是 阿尔茨海默病中的神经原纤维缠绕，肌营养不良中的球体 帕金森氏病的侧索硬化症和路易小体。尽管有一个或 更多的因素似乎会导致单体和 组成蛋白的非共价相关形式，主要是 神经丝(NF)或tau，申请人假定，并具有初步的 有证据表明，氧化应激过程会导致不可逆转的 蛋白质聚集体的“胶合”，至少部分是通过共价 交联剂。改性涉及直接氧化的组合 蛋白质侧链及其糖氧化产物和脂类加合物 预计将在重叠但不同的情况下表现为过氧化 三种疾病状态的标志物的模式。因此，了解 这些标志物的结构和程度在个体疾病中的差异 是为了帮助阐明疾病的发病机制和 为设计改良剂/方法提供线索。这项建议 代表了三个职业生涯中期的多学科合作 具有有机/多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(一) 交联型和非交联型氧化羰基化合物的结构解析 利用新型质谱学对蛋白质沉淀物的修饰 分析，(Ii)氧化特异性核因子和tau抗体的发展 构象表位和结构上定义的氧化损伤，以及 (3)大气污染物时空分布的相关性评价 不同脑区的氧化特异性标志物及其相互作用 病理生物学意义。

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.