Catalytic Activities of Ribozymes and DNAzymes in Water and Mixed Aqueous Media
Catalytic Activities of Ribozymes and DNAzymes in Water and Mixed Aqueous Media
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DOI：
10.3390/catal7120355
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发表时间：
2017-12-01
期刊：
影响因子：
3.9
通讯作者：
Sugimoto, Naoki
中科院分区：
文献类型：
--
作者：
Nakano, Shu-ichi;Horita, Masao;Sugimoto, Naoki
催化核酸被认为是潜在的治疗剂和生物传感器。核酸酶的催化活性通常在稀水溶液中进行研究，尽管活细胞内的反应环境的物理性质以及它们在其中操作的生物传感器的表面附近的区域中的物理性质与纯水的物理性质完全不同。分子环境的影响也是旨在通过向水溶液中添加有机溶剂来改善和扩展核酸功能的研究的重要焦点。在这项研究中，RNA和DNA酶（锤头状核酶，17 E DNAzyme，R3C核酶，和9DB1 DNAzyme）的催化活性进行了研究，使用21种不同的混合水溶液，包括有机化合物。动力学测量表明，相对于不含有机添加剂的溶液，这些酶在混合溶液中可以表现出增强的催化活性。相关性分析表明，在介电常数比水低的介质中，锤头状核酶催化的反应的周转率增加，而在增加DNA相互作用强度的条件下，17E DNAzyme催化的反应的周转率增加。而R3C核酶和9DB1 DNA酶没有表现出明显的周转活性，但在许多混合溶液中，它们的单周转率增加。我们的数据提供了在适用于医学和技术领域的各种条件下催化核酸的活性的见解，例如在活细胞和生物传感器中。
Catalytic nucleic acids are regarded as potential therapeutic agents and biosensors. The catalytic activities of nucleic acid enzymes are usually investigated in dilute aqueous solutions, although the physical properties of the reaction environment inside living cells and that in the area proximal to the surface of biosensors in which they operate are quite different from those of pure water. The effect of the molecular environment is also an important focus of research aimed at improving and expanding nucleic acid function by addition of organic solvents to aqueous solutions. In this study, the catalytic activities of RNA and DNA enzymes (hammerhead ribozyme, 17E DNAzyme, R3C ribozyme, and 9DB1 DNAzyme) were investigated using 21 different mixed aqueous solutions comprising organic compounds. Kinetic measurements indicated that these enzymes can display enhanced catalytic activity in mixed solutions with respect to the solution containing no organic additives. Correlation analyses revealed that the turnover rate of the reaction catalyzed by hammerhead ribozyme increased in a medium with a lower dielectric constant than water, and the turnover rate of the reaction catalyzed by 17E DNAzyme increased in conditions that increased the strength of DNA interactions. On the other hand, R3C ribozyme and 9DB1 DNAzyme displayed no significant turnover activity, but their single-turnover rates increased in many mixed solutions. Our data provide insight into the activity of catalytic nucleic acids under various conditions that are applicable to the medical and technology fields, such as in living cells and in biosensors.