Identification of post-translational modification in nuclear proteins during neural stem cell fate specification by proteomic analysis

通过蛋白质组学分析鉴定神经干细胞命运规范过程中核蛋白的翻译后修饰

• 批准号: 21770119
• 负责人: NIIMORI Kanako
• 金额: $ 2.91万
• 依托单位: Kumamoto University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Young Scientists (B)
• 财政年份: 2009
• 资助国家: 日本
• 起止时间: 2009 至 2010
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-21770119/
• 关键词: 神経幹細胞; プロテオミクス; 翻訳後修飾; リン酸化; 2D-DIGE; 分化制御; 転写調節因子; 質量分析

It is not yet completely understood how neural stem cell fate is specified, in spite of extensive investigations by, for instance, gene expression profiling. To tackle this important question, we have focused on the changes in post-translational modification of nuclear proteins such as transcription factors, coactivators/corepressors, and chromatin modifiers in neural stem cells (NSCs) in response to fibroblast growth factor 2 (FGF2), a representative factor to maintain them. NSCs were stimulated by FGF2, and its lysates were examined by proteomic analysis. The result revealed 80 up- or down-regulated protein spots upon FGF2 stimulation out of 4095 spots with statistically significant differences. Among them, we focused on 24 spots differentially phosphorylated in response to FGF2, and identified all of them by nanoLC-QqTOF MS analysis. These proteins contained factors related to nuclear dynamics including nuclear translocation and chromatin modifications.

尽管通过基因表达谱等进行了广泛的研究，但神经干细胞的命运是如何被指定的，目前还没有完全理解。为了解决这一重要问题，我们关注了神经干细胞（NSCs）中转录因子、共激活因子/共抑制因子和染色质修饰因子等核蛋白的翻译后修饰变化，以响应成纤维细胞生长因子2 (FGF2)，这是维持它们的代表性因子。用FGF2刺激NSCs，并通过蛋白质组学分析检测其裂解物。结果显示，在FGF2刺激下，4095个蛋白点中有80个上调或下调，差异有统计学意义。其中，我们重点选取了24个对FGF2响应的差异磷酸化位点，并通过nanoLC-QqTOF MS分析对其进行了鉴定。这些蛋白质含有与核动力学相关的因子，包括核易位和染色质修饰。