Department of Pharmacology and Experimental Neuroscience, University of Nebraska Medical Center, Omaha, Nebraska 68198, USA.
J Proteome Res. 2010 Jan;9(1):352-8. doi: 10.1021/pr900685u.
Investigating, predicting, diagnosing, and treating HIV-1-associated neurocognitive disorder (HAND) has been hindered by the lack of disease-related molecular markers. In this study, plasma from rhesus monkeys (n = 6), before and after infection with simian immunodeficiency virus (SIV), was profiled to obtain differential fingerprints in protein expression during SIV-induced central nervous system (CNS) disease. A quantitative proteomic analysis was performed by means of isobaric tag for relative and absolute quantification (iTRAQ) labeling, using multidimensional liquid chromatography tandem mass spectrometry (LC-MS/MS) run on a linear ion trap mass spectrometer in an integrated mode comprising pulsed-Q-dissociation (PQD) and CID. Among a panel of proteins showing differential expression following SIV infection, we identified afamin, a member of the albumin superfamily, to be significantly down regulated after infection. Validation by Western blot confirmed this observation and, given its potential implication in neuroprotection by transport of alpha-tocopherol (alphaTocH), provides new avenues into further understanding HIV induced CNS disease. iTRAQ-based LC-MS/MS provides a valuable platform for plasma protein profiling and has important implications in identifying molecular markers relevant for the pathogenesis of neurodegenerative diseases. Using such an approach, we show its successful application in identifying differential fingerprints in SIV/HIV induced CNS disease.
研究、预测、诊断和治疗 HIV-1 相关神经认知障碍 (HAND) 一直受到缺乏疾病相关分子标志物的阻碍。在这项研究中,对感染猴免疫缺陷病毒 (SIV)前后的恒河猴血浆(n = 6)进行了分析,以获得 SIV 诱导的中枢神经系统 (CNS) 疾病过程中蛋白质表达的差异指纹。通过使用等相对和绝对定量标记 (iTRAQ) 标记进行定量蛋白质组学分析,使用集成模式下的多维液相色谱串联质谱 (LC-MS/MS) 在线性离子阱质谱仪上运行,该模式包括脉冲 Q 解离 (PQD) 和 CID。在一组显示 SIV 感染后差异表达的蛋白质中,我们鉴定出 afamin,即白蛋白超家族的成员,在感染后显著下调。Western blot 验证证实了这一观察结果,并且鉴于其通过转运α-生育酚 (αTocH) 提供神经保护的潜在意义,为进一步了解 HIV 诱导的 CNS 疾病提供了新的途径。基于 iTRAQ 的 LC-MS/MS 为血浆蛋白质组学分析提供了一个有价值的平台,并在鉴定与神经退行性疾病发病机制相关的分子标志物方面具有重要意义。使用这种方法,我们展示了其在鉴定 SIV/HIV 诱导的 CNS 疾病差异指纹方面的成功应用。