用γ射线羟基自由基标记、质谱和分子对接等技术联合解析蛋白质复合物结构新技术的研究

项目名称： 用γ射线羟基自由基标记、质谱和分子对接等技术联合解析蛋白质复合物结构新技术的研究

项目编号： No.21277081

项目类型： 面上项目

立项/批准年度： 2013

项目学科： 环境科学、安全科学

项目作者： 刘汝涛

作者单位： 山东大学

项目金额： 80万元

中文摘要： 蛋白质复合物的结构解析是阐明细胞代谢、信号传导和基因表达等的关键环节，对环境污染与健康的研究具有重要意义。本研究拟通过γ射线照射，实现与溶剂水分子接触的蛋白质及复合物氨基酸侧链的氧化标记，然后联合生物酶切技术、LC-MS和分子对接技术建立解析蛋白质复合物结构可靠的新方法。解决联合解析技术中γ射线照射停止后溶液中发生继续氧化的难题；为生理条件下水溶液中微量(fmol-pmol)蛋白质复合物结构解析提供新的研究手段。利用该方法将解析p85ni:iSH2、TcP1beta:TcP2alpha等3-5个蛋白质复合物的未知结构；将拓宽γ射线的应用范围，提高我国在蛋白质复合物结构解析、分子水平上环境污染物生物学效应评价等领域研究的国际影响力。 解决上述学术问题，预计申请专利4-6项，发表SCI论文12-16篇，其中影响因子大于3的约7-10篇；培养5-6名硕士研究生、3-5名博士研究生，2-3名博士后

中文关键词： γ射线羟基自由基标记；质谱和光谱；蛋白质复合物结构；活性氧；毒性作用

英文摘要： The structure elucidation of protein-protein complex is the key network in elucidating the processes such as cellular metabolism, signal transduction and gene expression, and plays a significant role in the environmental pollution and health research. In this project, we utilize the γ-ray exposure to achieve the oxidation labeling of the solvent accessible side-chain of protein and protein-protein complexes, and then combine enzyme digestion technology, LC-MS and molecular docking method to establish the reliable and novel strategy in the structural assessment of protein-protein complex. We will solve the problems of the continue oxidation in solution after stopped radiation, and provide a new method to assess the structure of trace (fmol-pmol) complex under physiological conditions. This method will be put into use for analyze the undiscovered structures of 3-5 protein complex including p85ni:iSH2 and TcP1beta:TcP2alpha et.al, and extend the range of γ-ray application, which will also increase our international influence in the field of protein complex assessment and biological effect of environmental pollutants evaluation on molecular level. Solving the scientific problems, we will apply for 4-6 patents, publish 12-16 papers in SCI including 7-10 papers the impact factor of which exceed 3, and train 5-6 studen

英文关键词： Hydroxyl radical labeled after γ irradiation；MS and spectroscopy；protein complex；reactive oxygen species；toxic effect