采用比较蛋白质组学方法研究p38β激酶对293T细胞蛋白质表达的影响

Appling Comparative Proteomics Method to Study the Effect of p38β on the Expression of Proteins in Human Thymus Tumor 293T Cells

采用比较蛋白质组学方法研究了过表达p38信号通路激酶p38β对人类肿瘤细胞293T细胞蛋白质表达的影响,利用双向电泳和质谱技术分离鉴定了13个差异表达蛋白质.这些差异蛋白的分布从细胞质膜到核,其功能涉及核酸、蛋白质、糖脂类分子的转录、合成和成熟,细胞物质跨膜运输,细胞周期调控以及细胞防御等.文献分析并没有发现上述鉴定的差异表达蛋白是p38β激酶现有直接底物的证据,提示其可能为新的p38β激酶相关蛋白.这些结果表明,比较蛋白质组学方法是寻找激酶新靶标的一种有效工具.

The aim of the present study was to clarify the effect of p38β overexpression,a subtype of p38 MAP kinase,on the expression of proteins in human tumor cells for screening of its pathway-related proteins.Comparative proteomics methodologies were applied to screen the change of the expression of proteins in the human thymus tumor 293T cells responding to the overexpression of p38β kinase gene.Furthermore,the basic biological function and subcellular location of these changed proteins were analyzed by bioinformatics.13 differential expression proteins caused by the overexpression of p38β kinase gene were respectively separated and identified with two dimensional gel electrophoresis and mass spectrometry.The results show that these 13 proteins range from cellular membrane to cellular nuclear and take part in the cellular molecule synthesis,cellular transport across membrane and the regulation of the cell cycle.Bioinformatics analysis indicated that there was no the evidence that these proteins were the indirect substrates of p38β,suggesting that they would be novel related proteins of p38β.The current study suggests that comparative proteomics technique is an powerful tool to screen new target of kinase,which provides a clue to insight into the events of signaling transduction in human tumor cells.