Proteomic identification of oxidatively modified proteins in Alzheimer's disease brain. Part 1: Creatine kinase bb, glutamine synthase, and ubiquitin carboxy-terminal hydrolase L-1
Proteomic identification of oxidatively modified proteins in Alzheimer's disease brain. Part 1: Creatine kinase bb, glutamine synthase, and ubiquitin carboxy-terminal hydrolase L-1
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DOI：
10.1016/s0891-5849(02)00914-0
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发表时间：
2002-08-15
影响因子：
7.4
通讯作者：
Butterfield, DA
中科院分区：
文献类型：
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作者：
Castegna, A;Aksenov, M;Butterfield, DA
活性氧（ROS）对蛋白质的氧化改变与衰老以及阿尔茨海默病（AD）等年龄相关的神经退行性疾病的进展有关。蛋白质羰基是蛋白质氧化的一个标志物，在阿尔茨海默病患者的大脑中含量增加，这表明蛋白质的氧化修饰与阿尔茨海默病相关。氧化损伤可导致多种情况，例如特定蛋白质功能丧失、蛋白质清除异常、细胞氧化还原平衡的破坏以及对细胞周期的干扰，并最终导致神经元死亡。确定蛋白质氧化的特定靶点是建立阿尔茨海默病中氧化修饰与神经元死亡之间关系的关键步骤，我们实验室之前通过免疫化学检测肌酸激酶BB和β -肌动蛋白作为阿尔茨海默病大脑与对照大脑中特异性氧化的蛋白质，部分实现了这一目标。然而，这个过程很费力，需要有特定的抗体，而且最重要的是，首先需要对蛋白质的身份有一个合理的猜测。在这项研究中，我们提出了第一种蛋白质组学方法来鉴定阿尔茨海默病中特异性氧化的蛋白质，即通过将二维指纹图谱与羰基的免疫检测以及通过质谱鉴定蛋白质相结合。将蛋白质组学应用于神经退行性疾病所产生的强大技术揭示了阿尔茨海默病（AD）大脑中蛋白质氧化的特定靶点的存在：肌酸激酶BB、谷氨酰胺合成酶和泛素羧基末端水解酶L - 1。参考这些氧化修饰的蛋白质在阿尔茨海默病大脑神经退行性变中的潜在参与，对这些结果进行了讨论。蛋白质组学为鉴定衰老和年龄相关神经退行性疾病中氧化修饰的蛋白质提供了一种快速的方法，且没有免疫化学检测方法的局限性。（C）2002爱思唯尔科学公司
Oxidative alterations of proteins by reactive oxygen species (ROS) have been implicated in the progression of aging and age-related neurodegenerative disorders such as Alzheimer's disease (AD). Protein carbonyls, a marker of protein oxidation, are increased in AD brain, indicating that oxidative modification of proteins is relevant in AD. Oxidative damage can lead to several events such as loss in specific protein function, abnormal protein clearance, depletion of the cellular redox-balance and interference with the cell cycle, and, ultimately, to neuronal death. Identification of specific targets of protein oxidation represents a crucial step in establishing a relationship between oxidative modification and neuronal death in AD, and was partially achieved previously in our laboratory through immunochemical detection of creatine kinase BB and P-actin as specifically oxidized proteins in AD brain versus control brain. However, this process is laborious, requires the availability of specific antibodies, and, most importantly, requires a reasonable guess as to the identity of the protein in the first place. In this study, we present the first proteomics approach to identify specifically oxidized proteins in AD, by coupling 2D fingerprinting with immunological detection of carbonyls and identification of proteins by mass spectrometry. The powerful techniques, emerging from application of proteomics to neurodegenerative disease, reveal the presence of specific targets of protein oxidation in Alzheimer's disease (AD) brain: creatine kinase BB, glutamine synthase, and ubiquitin carboxy-terminal hydrolase L-1. These results are discussed with reference to potential involvement of these oxidatively modified proteins in neurodegeneration in AD brain. Proteomics offers a rapid means of identifying oxidatively modified proteins in aging and age-related neurodegenerative disorders without the limitations of the immunochernical detection method. (C) 2002 Elsevier Science Inc.