基于DNA折纸结构模板促进蛋白质结晶、纯化与分离

项目名称： 基于DNA折纸结构模板促进蛋白质结晶、纯化与分离

项目编号： No.21474059

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： 杨忠强

作者单位： 清华大学

项目金额： 88万元

中文摘要： 蛋白质的成核与结晶在解析蛋白质结构的基础研究以及蛋白质分离的药物应用中发挥着重要作用。通过尺寸效应，利用多孔表面对蛋白质进行异相成核结晶已有报道。然而，以往所用模板尺寸具有分散性、且难以在纳米尺度进行精确控制，限制蛋白质结晶过程的调控及机理研究。近年来迅速发展的DNA折纸结构展示出优于传统三维模板的特性：精确设计性、尺寸单一、纳米尺度连续可调、可定点定位修饰有机/无机/生物官能团等。在本申请中，我们将首次利用DNA折纸结构为模板，利用尺寸效应促进蛋白质结晶、纯化和分离。系统研究DNA折纸结构尺寸/形状对蛋白质结晶诱导时间、饱和浓度和晶体纯度的影响，以期提高结晶效率，降低结晶成本；借助模板结构精确可调控深入研究尺寸效应与蛋白结晶的关系和内在机理；尝试利用核酸适配体与蛋白的特异性相互作用，铆订目标蛋白实现其纯化和分离。该技术将开辟一个新的蛋白质结晶的平台，并对蛋白分离和工艺具有指导意义。

中文关键词： DNA折纸术；蛋白质；结晶；纳米材料；自组装

英文摘要： The protein nucleation and crystallization is playing an important role in not only revealing the protein structures in fundamental research but also separating proteins in pharmaceutical applications. Heterogeneous nucleation involving porous surfaces have been reported to crystallize proteins. However, it lacks of in-depth understanding and comprehensive investigations, mainly due to the difficulty of obtaining 3D nanotemplates with well-defined and tunable sizes and structures in nanoscale. In this proposal, we aim to explore DNA origami nanostructures to promote and control the nucleation and crystallization of proteins. This highly interdisciplinary work will be the first example of using DNA origami with nanoscale precision to crystallize proteins, elucidate how the size and shape of nanostructures influence induction time, supersaturated concentration and purity of proteins, and eventually by taking advantage of specific interaction of between DNA aptamer and proteins, investigating the crystallization of proteins which are difficult to crystallize or crystallization of target proteins from crude samples, even mixtures. The technique would open up a new platform of protein crystallization and benefit the bioseparation and bioprocessing for pharmaceutical companies and industries.

英文关键词： DNA origami;protein;crystallization;nanomaterial;self-assembly