Proteomic analysis of the mouse brain following protein enrichment by preparative electrophoresis
Proteomic analysis of the mouse brain following protein enrichment by preparative electrophoresis
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DOI：
10.1002/elps.200500562
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发表时间：
2006-04-01
期刊：
影响因子：
2.9
通讯作者：
Fountoulakis, M
中科院分区：
文献类型：
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作者：
Xixi, E;Dimitraki, P;Fountoulakis, M
蛋白质组学是研究大脑蛋白质的特性和水平的强大技术。蛋白质水平的变化以及神经系统疾病中发生的修饰可能为这些疾病的发病机制提供信息，并可能导致潜在药物靶点和疾病标志物的识别。为了提高表征复杂蛋白质谱的能力，应将蛋白质混合物分离成更简单的级分，从而增加检测低丰度蛋白质的可能性。考虑到低丰度蛋白质被认为参与重要的生物过程，识别这些低拷贝数基因产物似乎是一个科学挑战。在本研究中，通过制备型电泳富集后对成年小鼠脑组织进行蛋白质组分析。这是使用 PrepCell 装置在 0.1% 十二烷基硫酸锂存在下进行的。样品在圆柱形聚丙烯酰胺凝胶中进行电泳，收集的级分的蛋白质首先通过 1-D 分析，然后通过 2-DE 分析。通过MALDI-TOF-MS进行蛋白质鉴定。目前的分析鉴定出 360 种不同的基因产物。其中包括转运蛋白、转录激活剂、信号转导分子以及具有许多其他功能的蛋白质。制备型电泳是富集低分子量蛋白质的有效方法，可用于研究中枢神经系统疾病。
Proteomics is a powerful technology to study the identity and levels of brain proteins. Changes of protein levels as well as modifications that occur in neurological disorders may be informative for the pathogenesis of these disorders and could result in the identification of potential drug targets and disease markers. To increase the capability of characterizing complex protein profiles, protein mixtures should be separated into simpler fractions, thus increasing the likelihood of detecting low-abundance proteins. Considering that low-abundance proteins are thought to be involved in important biological processes, identification of those low-copy-number gene products appears to be a scientific challenge. In the present study, proteomic analysis of adult mouse brain tissue was performed following enrichment by preparative electrophoresis. This was performed using the PrepCell apparatus in the presence of 0.1 % lithium dodecyl sulfate. Samples were electrophoresed in a cylindrical polyacrylamide gel and the proteins of the fractions collected were first analyzed by 1-D and then by 2-DE. Protein identification was performed by MALDI-TOF-MS. The present analysis resulted in the identification of 360 different gene products. Among those were transport proteins, transcription activators, signal transduction molecules as well as proteins with a number of other functions. Preparative electrophoresis is an efficient method for the enrichment of proteins of low molecular mass and may be useful in the investigation of disorders of the central nervous system.