基于iTRAQ的鸟枪法神经蛋白质组学
iTRAQ-based shotgun neuroproteomics.
作者信息
Liu Tong, Hu Jun, Li Hong
机构信息
Department of Biochemistry and Molecular Biology, University of Medicine and Dentistry of New Jersey, Newark, NJ, USA.
出版信息
Methods Mol Biol. 2009;566:201-16. doi: 10.1007/978-1-59745-562-6_14.
Abstract
Shotgun proteomics involves the analysis of peptides obtained by enzymatic digestions of proteins and subsequent identification via tandem mass spectrometry. This approach is an effective method for studying global protein expression in neuronal systems. The method described here is a quantitative shotgun neuroproteomics method using amine-specific isobaric tags for a relative and absolute quantitation (iTRAQ)-based workflow. We will provide the technical details for sample preparation, two-dimensional liquid chromatography, tandem mass spectrometry, database search, and statistical analysis to identify differentially expressed proteins. We will use a recent study on a rat model of multiple sclerosis, experimental autoimmune encephalomyelitis to illustrate the successful application of this method.
摘要
鸟枪法蛋白质组学涉及对通过蛋白质酶解获得的肽段进行分析，并随后通过串联质谱进行鉴定。这种方法是研究神经元系统中整体蛋白质表达的有效手段。这里所描述的方法是一种基于胺特异性等压标签相对和绝对定量（iTRAQ）工作流程的定量鸟枪法神经蛋白质组学方法。我们将提供样品制备、二维液相色谱、串联质谱、数据库搜索以及统计分析的技术细节，以鉴定差异表达的蛋白质。我们将利用最近一项关于多发性硬化症大鼠模型——实验性自身免疫性脑脊髓炎的研究来说明该方法的成功应用。
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