调控Keap1与Nrf2相互作用的Keap1蛋白未知磷酸化位点及磷酸化信号传导通路研究

项目名称： 调控Keap1与Nrf2相互作用的Keap1蛋白未知磷酸化位点及磷酸化信号传导通路研究

项目编号： No.31460226

项目类型： 地区科学基金项目

立项/批准年度： 2015

项目学科： 生物科学

项目作者： 王大勇

作者单位： 海南大学

项目金额： 53万元

中文摘要： Keap1蛋白对细胞内Nrf2蛋白水平的调节是生物体内调节氧化还原稳态水平的主要方式。转录因子Nrf2通过与抗氧化反应元件（ARE）结合，诱导100多种基因的表达；Nrf2与其胞内抑制物Keap1结合后迅即发生泛素依赖性降解。现有研究表明，Keap1与Nrf2的分离过程对氧化应激不敏感，可能需要细胞内的蛋白质磷酸化信号系统参与调控Keap1与Nrf2的相互作用，但目前尚未发现细胞内磷酸化信号系统调节Keap1与Nrf2相互作用的方式。本项目假设，Keap1蛋白上的特定氨基酸位点磷酸化可以调节Keap1与Nrf2的相互结合，从而影响Nrf2-ARE信号传导。本项目采用分子生物学、质谱分析和生物化学等直接的实验方法研究分析具有上述调节功能的人Keap1磷酸化位点及主要信号传导通路，为抗氧化、抗炎、组织再生、生长发育、预防肿瘤、抗衰老、以及老年痴呆等基础和应用研究提供确切的依据。

中文关键词： 蛋白质相互作用；Keap1；磷酸化；信号转导；氧化应激

英文摘要： The regulation of Keap1 on intracellular level of Nrf2 is the most important mechanism regulating redox homeostasis in a living organism. Nrf2， which is a transcription factor, regulates the expression of more than 100 genes by binding to antioxidant response element (ARE); it is rapidly degradated in a ubiquitin-dependent manner after binding to its endogenous inhibitor Keap1.The separation of Keap1 from Nrf2 is not sensitive to oxidative stress, and intracellular phosphorylation-signaling system may involved in regulating the separation, however, the signal regulating process has not been elucidated yet. It is hypothesized in the proposal that there are special amino residues that can be phosphorylated to regulate the binding of Keap1 to Nrf2, which in turn influence the signal transduction from Nrf2 to ARE. In the proposal, direct approaches involving molecular biological, mass spectrometric and biochemical methods will be employed to identify the phosphorylation sites on Keap1 and its upstream signaling pathways. The study will benefit basic and new medicine-related studies on antioxidation,anti-inflammation, tissue regeneration, growth and development, prevention of carcinogenesis, anti-aging, and prevention of Alzheimer's diseases.

英文关键词： Protein interaction;Keap1;Phosphorylation;Signal transduction;Oxidative stress