Mello C F, Sultana R, Piroddi M, Cai J, Pierce W M, Klein J B, Butterfield D A
Department of Chemistry, Center of Membrane Sciences and Sanders-Brown Center on Aging, University of Kentucky, Lexington, KY 40506, USA.
Neuroscience. 2007 Jul 13;147(3):674-9. doi: 10.1016/j.neuroscience.2007.04.003. Epub 2007 Jun 14.
Acrolein, the most reactive of the alpha,beta-unsaturated aldehydes, is endogenously produced by lipid peroxidation, and has been found increased in the brain of patients with Alzheimer's disease. Although it is known that acrolein increases total protein carbonylation and impairs the function of selected proteins, no study has addressed which proteins are selectively carbonylated by this aldehyde. In this study we investigated the effect of increasing concentrations of acrolein (0, 0.005, 0.05, 0.5, 5, 50 microM) on protein carbonylation in gerbil synaptosomes. In addition, we applied proteomics to identify synaptosomal proteins that were selectively carbonylated by 0.5 microM acrolein. Acrolein increased total protein carbonylation in a dose-dependent manner. Proteomic analysis (two-dimensional electrophoresis followed by mass spectrometry) revealed that tropomyosin-3-gamma isoform 2, tropomyosin-5, beta-actin, mitochondrial Tu translation elongation factor (EF-Tu(mt)) and voltage-dependent anion channel (VDAC) were significantly carbonylated by acrolein. Consistent with the proteomics studies that have identified specifically oxidized proteins in Alzheimer's disease (AD) brain, the proteins identified in this study are involved in a wide variety of cellular functions including energy metabolism, neurotransmission, protein synthesis, and cytoskeletal integrity. Our results suggest that acrolein may significantly contribute to oxidative damage in AD brain.
丙烯醛是α,β-不饱和醛中反应活性最强的一种，由脂质过氧化作用内源性产生，且在阿尔茨海默病患者的大脑中含量升高。尽管已知丙烯醛会增加蛋白质的总羰基化水平并损害某些特定蛋白质的功能，但尚无研究探讨哪些蛋白质会被这种醛选择性地羰基化。在本研究中，我们调查了不同浓度（0、0.005、0.05、0.5、5、50微摩尔）的丙烯醛对沙鼠突触体中蛋白质羰基化的影响。此外，我们运用蛋白质组学方法来鉴定被0.5微摩尔丙烯醛选择性羰基化的突触体蛋白。丙烯醛以剂量依赖的方式增加蛋白质的总羰基化水平。蛋白质组学分析（二维电泳后进行质谱分析）显示，原肌球蛋白-3-γ同工型2、原肌球蛋白-5、β-肌动蛋白、线粒体Tu翻译延伸因子（EF-Tu(mt)）和电压依赖性阴离子通道（VDAC）被丙烯醛显著羰基化。与在阿尔茨海默病（AD）大脑中鉴定出特定氧化蛋白质的蛋白质组学研究一致，本研究中鉴定出的蛋白质参与了包括能量代谢、神经传递、蛋白质合成和细胞骨架完整性在内的多种细胞功能。我们的结果表明，丙烯醛可能对AD大脑中的氧化损伤有显著影响。
