催化活性DNA寡聚物(或DNA酶)提供了广泛的功能和应用。虽然已经知道了二十多年,DNA酶的作用模式仍然知之甚少,主要是由于缺乏高分辨率的结构见解。由于它们的分子大小,结构灵活性,以及与金属离子辅因子的动态相互作用,溶液核磁共振波谱(NMR)可以作为获得DNAzyme机理见解的最佳工具。在这方面,在催化循环过程中,几乎所有的DNA酶和它的底物状态都是可以达到的。下文中提供的说明书和方案可以辅助DNA酶的基于NMR的表征的初始步骤。为了减少初始设置要求并促进令人兴奋的新研究项目,所讨论的方法侧重于不需要成本密集型同位素标记策略的实验。
Catalytically active DNA oligomers (or
DNAzymes) offer a broad spectrum of functions as well as applications. Although known for over two decades, the DNAzyme\'s mode-of-actions are still poorly understood, mainly due to lack of high-resolution structural insights. Due to their molecular size, structural flexibility, and dynamic interactions with metal-ion cofactors, solution nuclear magnetic resonance spectroscopy (NMR) can serve as optimal tool to obtain mechanistic insights of
DNAzymes. In this respect, nearly all states of the DNAzyme and its substrate during the catalytic cycle are accessible. The instructions and protocols provided in the following may assist the initial steps of an NMR-based characterization of
DNAzymes. To reduce the initial setup requirements and foster exciting new research projects, the discussed approaches focus on experiments that do not require cost-intensive isotope labeling strategies.