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特征的研究 与衰老和年龄相关的神经变性有关， 负责蛋白质沉积、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纤维缠结，肌萎缩性侧索硬化症中的球状体， 脊髓侧索硬化症和帕金森氏病中的路易体。 尽管一个或 更多的因素似乎诱导单体和 组成蛋白质的非共价结合形式，主要是 申请人假设，神经丝蛋白（NF）或tau蛋白是一种特异性的蛋白质，并且具有初步的 证据表明，氧化应激过程导致不可逆的 蛋白质聚集体的“粘合”，至少部分地通过共价键， 交联 修饰包括直接氧化 蛋白质侧链和加合的产物的糖氧化和脂质 预计将在重叠但不同的 三种疾病状态的标志物模式。 因此， 这些标记物的结构和程度的个体疾病变化 是假设，以帮助澄清疾病的发病机制， 为设计改良剂/方法提供线索。 该提案 代表了三个职业中期的多学科合作 具有有机/肽化学专业知识的研究人员（塞尔）， 蛋白质/分析生物化学（安德森），和 神经病理学/免疫生物化学（佩里），目的是定义 不可逆蛋白质沉积的结构基础。 目标是（一） 交联和非交联羰基氧化物的结构解析 使用新的质谱生物化学方法对蛋白质沉积物进行修饰 分析，（ii）开发氧化特异性NF和tau抗体 构象表位和结构定义的氧化损伤，和 (iii)时空分布的相关性评价 不同脑区的氧化特异性标记物及其 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.