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首页> 外文期刊>Mass Spectrometry Letters >Differential Protein Quantitation in Mouse Neuronal Cell Lines using Amine-Reactive Isobaric Tagging Reagents with Tandem Mass Spectrometry
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Differential Protein Quantitation in Mouse Neuronal Cell Lines using Amine-Reactive Isobaric Tagging Reagents with Tandem Mass Spectrometry

机译：小鼠神经元细胞系中使用胺反应性等压标记试剂和串联质谱的蛋白质差异定量分析

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The high-throughput identification and accurate quantification of proteins are essential strategies for exploring cellular functions and processes in quantitative proteomics. Stable isotope tagging is a key technique in quantitative proteomic research, accompanied by automated tandem mass spectrometry. For the differential proteome analysis of mouse neuronal cell lines, we used a multiplexed isobaric tagging method, in which a four-plex set of amine-reactive isobaric tags are available for peptide derivatization. Using the four-plex set of isobaric tag for relative and absolute quantitation (iTRAQ) reagents, we analyzed the differential proteome in several stroke time pathways (0, 4, and 8 h) after the mouse neuronal cells have been stressed using a glutamate oxidant. In order to obtain a list of the differentially expressed proteins, we selected those proteins which had apparently changed significantly during the stress test. With 95% of the peptides showing only a small variation in quantity before and after the test, we obtained a list of eight up-regulated and four down-regulated proteins for the stroke time pathways. To validate the iTRAQ approach, we studied the use of oxidant stresses for mouse neuronal cell samples that have shown differential proteome in several stroke time pathways (0, 4, and 8 h). Results suggest that histone H1 might be the key protein in the oxidative injury caused by glutamate-induced cytotoxicity in HT22 cells.
机译：高通量鉴定和准确定量蛋白质是探索定量蛋白质组学中细胞功能和过程的基本策略。稳定的同位素标记是定量蛋白质组学研究中的一项关键技术，并伴有自动串联质谱。对于小鼠神经元细胞系的差异蛋白质组分析，我们使用了多重等压标记方法，其中四组胺反应性等压标记可用于肽衍生化。使用四重等压标记的相对和绝对定量（iTRAQ）试剂组，我们在使用谷氨酸氧化剂对小鼠神经元细胞施加压力后，分析了几种中风时间通路（0、4和8 h）中的差异蛋白质组。 。为了获得差异表达蛋白质的列表，我们选择了在压力测试过程中明显改变的蛋白质。在测试之前和之后，由于95％的肽在数量上仅表现出很小的变化，我们获得了中风时间通路中八种上调和四种下调的蛋白质的列表。为了验证iTRAQ方法，我们研究了氧化应激在小鼠神经元细胞样品中的使用，这些样品在几个中风时间路径（0、4和8 h）中显示出差异蛋白质组。结果提示，组蛋白H1可能是谷氨酸诱导的HT22细胞毒性引起的氧化损伤的关键蛋白。

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