时间分辨共振拉曼光谱用于光感蛋白SRII/HtrII复合物的弱相互作用研究

项目名称： 时间分辨共振拉曼光谱用于光感蛋白SRII/HtrII复合物的弱相互作用研究

项目编号： No.21473161

项目类型： 面上项目

立项/批准年度： 2015

项目学科： 数理科学和化学

项目作者： 王惠钢

作者单位： 浙江理工大学

项目金额： 90万元

中文摘要： 本项目以掌控光感蛋白SRII/HtrII 复合物的光感信号传输以及氢键等弱相互作用变换规律为目的，拟采用稳态及时间分辨紫外共振拉曼光谱（UVRR）技术，结合三级联光谱仪对SRII/HtrII 光诱导信号传输过程进行研究。以基因定点突变技术以及同位素交换实验为基础进行振动光谱指认，选取225nm 为探测波长研究芳基侧链、氢键以及溶剂与侧链间的相互作用，重点分析光诱导过程SRII 和HtrII 接触区的氢键变化规律；研究氨基骨架及孤电子对跃迁增强的氢键信息，根据氨基振动信息表征HAMP 区域dHAMP 和cHAMP 构型的平衡转换。通过本项目研究希望为生物自组装体系弱相互作用的表征引入光谱方法，探索SRII/HtrII 复合物光感信号传输过程动力学规律，为氢键等弱相互作用和质子转移理论的发展构筑基础数据。

中文关键词： 时间分辨共振拉曼光谱；反应动力学；SRII/HtrII；复合物；弱相互作用；羰基振动

英文摘要： The motivations for this proposal is to better understand the photo-induced signal transduction and the consequent structure changes around the hydrogen bonding and the other weak interactions. Holding this popurse, our investigations will carried out on the study of photo-induced signal transduction and conformation changes in the photoreceptor/transducer complex SRII/HtrII using Steady-state and time resolved ultraviolet resonance Raman (UVRR) spectroscopy as well as TriVista monochromator. Spectra assignments are based on mutants as reference systems and H/D exchange spectra. UVRR spectroscopy can serve as a probe of the aromatic side-chain environment (~225 nm laser excitation) and the amide backbone conformation. The 225 nm UVRR spectra show selective enhancements of the aromatic side-chains, whose intensities and frequencies strongly depend on hydrogen bonding and solvent access to the side chains. We will use nanosecond time resolved deep UVRR spectroscopy with 225 nm laser excitation to determine changes in hydrogen bonding of aromatic amino acids in the contact region of the SRII/HtrII complex. Further, UVRR spectroscopy will be employed to characterize the enhanced hydrogen bonding spectra and equilibrium between the dynamic (dHAMP) and compact (cHAMP) form of the HAMP domain in HtrII via the changes of the amide modes. This proposal intends to provide the time resolved Resonance Raman Spectra as a powerful in-situ detection tool for the research of weak interactions in the self-assemble systems. We will explore the photo-induced signal transduction and conformational dynamics in the photoreceptor/transducer complex SRII/HtrII. Hope that this research will pave the way for the development of weak interaction and proton transfer theory.

英文关键词： time resolved resonance Raman spectra;reaction dynamic;SRII/HtrII complex;weak interaction;C=O vibration