基于表面增强共振拉曼光谱技术的脱氧核酶构效关系研究

项目名称： 基于表面增强共振拉曼光谱技术的脱氧核酶构效关系研究

项目编号： No.30870533

项目类型： 面上项目

立项/批准年度： 2009

项目学科： 生物科学

项目作者： 李正强

作者单位： 吉林大学

项目金额： 32万元

中文摘要： 脱氧核酶的动态结构与脱氧核酶活性的关系研究对于了解和掌握核酶催化性质变化的内在规律、设计在近生理条件下高度底物识别和高度切割效率的新型核酶等具有重要的意义。目前，研究生物大分子结构的技术手段主要集中在X 射线衍射和NMR 上，它们在核酸研究方面具有一定的局限性。本项目旨在利用表面增强共振拉曼光谱（SERRS）技术对系列脱氧核酶（不同结合臂，突变/修饰碱基）环境变化状态下的自身动态结构以及脱氧核酶切割底物的动态过程进行检测，以获得的结构信息结合放射自显影技术得到的脱氧核酶的切割效率来阐述脱氧核酶的自身结构变化对其切割效率的影响，解释反应过程中催化中心各碱基的作用机制，总结在近生理条件下切割效率较高的新型脱氧核酶的设计原则，最终为设计高度底物识别和高度切割效率的新型脱氧核酶提供理论。

中文关键词： 核酶;脱氧核酶;表面增强共振拉曼光谱;动态结构;催化活性

英文摘要： Ribozymes and DNAzyme are important fields in enzymology.The structural biology of ribozymes and DNAzyme is significant in design of ribozymes or DNAzyme with high identify to substrate and catalytic activity. Now,the main ways in studying structurefunction relationships of biological macromolecule is NMR and x-ray diffraction technique,but they are limited to acquire the true relation between dynamic structure and function of nucleic acids. Raman spectroscopy is a valuable tool in various research fields. The technique yields structural information from all kind of samples often without the need for extensive sample preparation.Moreover,Surface Enhanced Raman Scattering (SERS) gives rise to analytical applications with much promise.This project is aimed to understand the effect of the change of environment on the dynamic structure of a DNAzyme and the dynamic formation process of the DNAzyme -substrate complex by the way of surface-enhanced raman scattering (SERS). Then,combining with the catalytic activity of the DNAzyme from the autoradiography experiment, the structure-function relationships of the DNAzyme, the catalytic mechanism and the function of the every single base in the DNAzyme catalytic process can be inferred.

英文关键词： Ribozyme;DNAzyme;Surface Enhanced Raman Scattering (SERS); Dynamic structure;Catalytic activity