膜蛋白Leptosphaeria rhodopsin二聚化组装的结构机制研究

项目名称： 膜蛋白Leptosphaeria rhodopsin二聚化组装的结构机制研究

项目编号： No.21273279

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： 陈德亮

作者单位： 中国科学院大学

项目金额： 68万元

中文摘要： Leptosphaeria rhodopsin是真核生物中发现的一种七次螺旋跨膜的视黄醛蛋白，以特殊稳定的二聚体形式组装，对膜蛋白组装研究具有重要的模型意义。本项目拟从Leptosphaeria Rhodopsin跨膜螺旋结构入手，主要运用空间定位顺磁共振技术，确认参与二聚化组装的螺旋及其相对空间位置；同时进行Leptosphaeria rhodopsin蛋白晶体的培养优化，直接解析出分子间相互作用的螺旋结构与氨基酸残基。根据结构数据进一步设计突变体蛋白，运用圆二色谱等技术表征二聚体组装的程度；以光解实验和顺磁共振分别表征蛋白稳定性与构象均一性。从而初步阐明Leptosphaeria Rhodopsin二聚化组装的结构机制。

中文关键词： 膜蛋白；寡聚化；菌紫质；；

英文摘要： Leptosphaeria rhodopsin is a eukaryotic and seven helical transmembrane retinal protein. It forms as a stable dimer and may contribute as an important model of membrane protein assembly. We plan to use mainly the site directed electron magnetic resonance technique to identify the helices which contribute to the dimerization, and their relative spatial locations. Meanwhile, we are trying to optimize the crystal quality and hopefully to solve the dimer structure, therefore, directly to identify those residues within the dimer interface. With the structural data in hand, we plan to design new Leptosphaeria rhodopsin mutants, so that we will characterize the dimerization extent by circular dichroism. Furthermore, we will calculate the stability and conformational homogeneity of Leptosphaeria rhodopsin by the light induced denaturation experiment and electron paramagnetic resonance, respectively. Finally, we are trying to illustrate the structural mechanism of the dimerization assembly in Leptosphaeria rhodopsin.

英文关键词： membrane protein；oligomization；bacteriorhodopsin；；