葫芦脲和药物与生物大分子相互作用及分析应用

【作者】 吴霞；

【导师】 杨景和；

【作者基本信息】 山东大学 ， 分析化学， 2006， 博士

【摘要】 蛋白质和核酸是两类重要的生物大分子。蛋白质担负着各种生理功能，从整体上维持生物体新陈代谢活动的进行，是生物性状的直接表达者，而核酸是揭示生命遗传与变异的主要物质。因此，深入研究小分子物质与蛋白质和核酸的作用机理，建立蛋白质和核酸快速、简便的定量分析方法，对在分子水平上阐明生命的奥秘、开发新药和预防与治疗疑难病症等方面具有重要的意义，是当前生物分析化学研究的前沿和热点之一。 本论文以分析化学，生物化学为研究背景，利用荧光技术、共振光散射技术、吸收光谱技术、圆二色光谱技术、透射电镜技术和扫描电子显微镜技术等手段研究了葫芦脲、药物与蛋白质和核酸的相互作用机理。利用荧光、共振光散射技术建立了蛋白质和核酸快速、准确、简便、灵敏的定量分析方法。论文共分六个部分。 论文的第一部分评述了近年来蛋白质的荧光和共振光散射探针以及核酸的荧光探针的研究进展；并对新一代超分子主体化合物葫芦脲的分子识别作用以及利用荧光和吸收等光分析手段对葫芦脲与有机小分子相互作用的研究进行了综述。共引用文献188篇。 本论文的第二部分，研究了酸性条件下，葫芦脲对ANS-BSA的荧光增强效应，建立了灵敏的定量测定蛋白质的新方法，BSA的线性范围5.0×10^-85.0×10^-6g／mL，检出限（S／N=3）为34ng／mL。将该方法用于人血清总蛋白的测定，结果令人满意。利用吸收光谱、园二色光谱、共振光散射光谱、荧光偏振、荧光寿命以及F（?）rster能量转移理论等对体系的作用机理进行了研究。研究表明，CB使得BSA的疏水微区暴露出来，更易于与ANS结合，从而促进了BSA吸收的能量向ANS转移，导致其荧光寿命的增加和量子产率的提高；CB和ANS的协同作用能导致BSA有强的去折叠作用。基于上述研究，提出了CB-ANS-BSA夹心式结构模型，即ANS结合在BSA的疏水微区后，再以分子间作用力结合CB，形成了CB-ANS-BSA夹心式结构的聚集体。 论文第三部分合成了新的纳米微粒葫芦脲银，并将其用于蛋白质的定量测定。利用柠檬酸钠还原法制备了银纳米粒子（nanoAg），并合成了新的纳更多还原

【Abstract】 Proteins and nucleic acids are very important bio-macromolecules. Different proteins perform a wide variety of biological functions. Some proteins are enzymes, which catalyze chemical reactions. Other proteins play structural or mechanical roles, such as maintenance the metabolism of plants and animals directly express biologic characters. Nucleic acids play a central role in the storage replication of hereditary and aberrance information and in the expression of this information through synthesis. In order to surmount many difficult diseases and explore the secret of life, we further investigate the interaction between small molecules and nucleic acids or proteins. A rapid and convenient method is discovered for quantitatively analyzing nucleic acids and proteins.Based on the research of analytical chemistry and biochemistry, this dissertation investigated the interaction mechanism between small molecules, supermolecules or nanoparticles and proteins as well as small molecules and nucleic acids using the multi-techniques including fluorescence, resonance light scattering, absorption, Circular Dichroism, Transmission Electron Microscope and Scanning Electron Microscope. Several rapid, efficient methods, which are of high sensitivity, are developed for determining proteins and nucleic acids. This dissertation is written in six chapters.The progress of fluorescence probes and resonance light scattering probes for proteins is summarized. The progress of fluorescence probes for nucleic acids is recapitulated. The new macromolecule host compound cucurbituril (CB) and the interaction between CB and metal ions or small organic molecules were reviewed in the first chapter. References (188) are cited here.Second chapter describes the properties and the scope of application of CB. It is found that CB can enhance the fluorescence of 1,8-ANS-BSA system in更多还原