核糖体蛋白RPS5与抗肝纤维化药物复合物的结构研究

项目名称： 核糖体蛋白RPS5与抗肝纤维化药物复合物的结构研究

项目编号： No.21273283

项目类型： 面上项目

立项/批准年度： 2013

项目学科： 数理科学和化学

项目作者： 仲维清

作者单位： 中国人民解放军第二军医大学

项目金额： 82万元

中文摘要： 我们近期研究发现核糖体蛋白RPS5在肝纤维化中起重要作用：具有抗肝纤维化作用的苦参碱及其衍生物M19以及木犀草素（Lut）等能特异性结合RPS5，药理实验证明这些化合物至少部分通过RPS5发挥抗肝纤维化作用，即RPS5是肝纤维化治疗的新靶点。但是，这些化合物与RPS5的结合位点仍为空白。本项目拟通过表达纯化出人的RPS5，研究RPS5与M19、Lut等化合物的结合性质，并培养两者复合物晶体，解析其晶体结构，分析其结合位点，再对RPS5进行突变，研究蛋白突变体与化合物的结合性质并确定其关键结合位点；在此基础上，构建以RPS5为靶点的结构模型，设计M19和Lut的新型衍生物。研究结果可在分子结构基础上阐明这些化合物的抗肝纤维化机制，并为以RPS5作靶点设计新型抗肝纤维化药物提供理论和实验依据。

中文关键词： 核糖体蛋白S5；木犀草素；结合位点；抗肝纤维化；作用机制

英文摘要： Our resent researches was initiated to address that ribosomal protein 5S （RPS5） plays a critical role in hepatic fibrosis. Matrine, its derivative M19 and luteolin, which occurred anti-hepatic fibrosis activity, can specially bind to RPS5, and the pharmacological experiments confirmed that RPS5 participates in the anti-hepatic fibrosis process of those compounds in a certain pattern. These results indicate that RPS5 is one of the new targets for treatment of hepatic fibrosis. However, the binding sites of RPS5 to these compounds are not clear. In this project, we are going to (1) express, isolate and purify the human RPS5 protein, and investigate the binding properties of RPS5 with M19 and luteolin；（2） fix out the binding sites by solving the crystal structures of the RPS5-M19/luteolin adducts，(3) determine the key sites involved in the interaction between RPS5 and M19/luteolin by comparing the binding properties of RPS5 mutants to M19/luteotin with those of native protein, and (4) establish molecule structural model to design new derivatives of M19 and luteolin against hepatic fibrosis on the target of RPS5. The research would illustrate the mechanism of matrine derivatives/luteotin against hepatic fibrosis, and collect the theoretical and experimental evidences on RPS5 acting as the new target for newly design

英文关键词： ribosomal protein S5；luteolin；binding sites；anti-hepatic fibrosis；reaction mechanism