THE STUDY OF PROTEIN DAMAGE IN AGING AND AGE-RELATED DISEASES IN BRAIN USING BI

使用 BI 研究大脑衰老和年龄相关疾病中的蛋白质损伤

• 批准号: 8168138
• 负责人: June Feng
• 金额: $ 5.55万
• 依托单位: LOUISIANA STATE UNIV A&M COL BATON ROUGE
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-07-15 至 2011-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8168138
• 关键词: Adenocarcinoma Cell; Affect; Affinity; Aging; Alzheimer' s Disease; Appearance; Binding; Brain; Colon Adenocarcinoma; Complex; Computer Retrieval of Information on Scientific Projects Database; Development; Devices; Disease; Disease Progression; Dose; Fluorescence Microscopy; Funding; Future; Goals; Grant; Human; Hydrogen Peroxide; Impaired cognition; In Vitro; Institution; Label; Laboratories; Liver; Mitochondrial Proteins; Modeling; Molds; Neurologic; Nickel; Oxidative Stress; Pathogenesis; Pathway interactions; Process; Protein Microchips; Proteins; Proteomics; Reactive Oxygen Species; Relative (related person); Research; Research Personnel; Resources; Sampling; Source; Sprague-Dawley Rats; Staging; Surface; Transgenic Mice; United States National Institutes of Health; age related; aging brain; amyloid pathology; base; microchip; mouse model; oxidation; oxidative damage

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. Oxidative stress such as protein carbonylation, nitrosylation and glutathionylation formation, induced by reactive oxygen species has been implicated as a contributing factor to Alzheimer Disease (AD). They are considered markers of oxidative stress in aging and AD. The overall goal of this research is to identify the major carbonylated and nitorsylated proteins in the aging brain that contribute to the pathogenesis of AD and determine which neurological pathways are affected by the presence of these oxidative damaged proteins. Our first specific aim is to develop a "micro-fluidic chip" to enrich a minute amount of damaged proteins prior to proteomics. The microchip is replicated from a nickel mold master using hot embossing. This surface of microchip is chemically modified to specifically bind to carbonyls. The capture of the protein carbonyls is further validated with in-chip protein labeling and fluorescence microscopy. This microchip based protein carbonyl enrichment proves to be a very sensitive and specific affinity device to enrich protein carbonyls from minuscule samples. It can detect and capture 40 femtomole of carbonlys. Our second specific aim is to develop a proteomic reactor to simplify the processing of complex proteomic samples by combining multiple proteomic steps and allow for the identification of carbonylated, nitrosylated and glutathionylated proteins. This will help to understand the longitudinal change in the identities and relative ratios of specific protein targets of oxidative damage. Liver mitochondrial proteins from Sprague Dawley rats subjected to hydrogen peroxide oxidation are used to confirm the feasibility of quantitative identification of carbonylated proteins using the proteomic reactor and iTRAQ labeling. Additionally, we identify mitochondrial proteins susceptible to carbonylation and nitrosylation in a dose-dependent manner from In vitro oxidative stress model-HT29 human colon adenocarcinoma cell with menodione treatment using quantitative proteomics. In the future study, to understand the early and progressive cellular changes in AD development and progression, using quantitative proteomic profiling, this laboratory will investigate protein carbonylation in the AD transgenic mouse model at three stages of disease progression: long before (2-months old), immediately before (6-months old) and after (12-months old), the appearance of amyloid pathology and cognitive impairment.

该副本是利用众多研究子项目之一 由NIH/NCRR资助的中心赠款提供的资源。子弹和 调查员（PI）可能已经从其他NIH来源获得了主要资金， 因此可以在其他清晰的条目中代表。列出的机构是 对于中心，这不一定是调查员的机构。 由活性氧诱导的氧化应激，例如蛋白羰基化，亚硝基化和谷胱甘肽形成，已被涉及成为阿尔茨海默氏病（AD）的促成因素。它们被认为是衰老和AD中氧化应激的标志物。这项研究的总体目的是确定衰老大脑中主要的羰基和硝基化蛋白，这些蛋白有助于AD的发病机理，并确定哪些神经系统途径受这些氧化受损蛋白的存在影响。 我们的第一个具体目的是开发“微氟芯片”，以在蛋白质组学之前富集一定量的受损蛋白质。使用热压器从镍模具主复制微芯片。这种微芯片的表面被化学修饰，以特异性结合羰基。通过芯片内蛋白质标记和荧光显微镜进一步验证蛋白羰基的捕获。这种基于微芯片的蛋白羰基富集被证明是一种非常敏感且特异性的亲和力装置，可从微小样品中富集蛋白质羰基。它可以检测并捕获40个碳液含量。 我们的第二个具体目的是开发蛋白质组学反应器，通过组合多个蛋白质组学步骤来简化复杂的蛋白质组学样品的处理，并允许鉴定羰基化，硝基化和谷胱甘肽的蛋白质。这将有助于理解氧化损伤的特定蛋白质靶标的身份和相对比的纵向变化。来自Sprague Dawley大鼠的肝脏线粒体蛋白使用过氧化氢氧化的大鼠使用蛋白质组织反应器和ITRAQ标记来确认羰基化蛋白定量鉴定的可行性。此外，我们通过使用定量蛋白质组学从体外氧化应激模型中鉴定出剂量依赖性的线粒体蛋白，并以剂量​​依赖性的方式依赖剂量依赖性方式，并通过细蛋白二酮治疗含量的体外氧化应激模型-HT29人类结肠腺癌细胞。 在未来的研究中，要使用定量的蛋白质组学分析来了解AD发育和进展的早期和进行性细胞变化，该实验室将在疾病进展的三个阶段研究AD转基因小鼠模型中的蛋白质羰基化：早在（2个月大）之前（2个月），在（6个月旧）和（12个月旧）和amby pathology and Pathology and Pathology and Pathology and Pathibolicagy and pathibolicy and andolody and andolody and。