De novo synthesis and development of an RNA enzyme
De novo synthesis and development of an RNA enzyme
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DOI：
10.1073/pnas.0405886101
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发表时间：
2004-09-21
影响因子：
11.1
通讯作者：
Inoue, T
中科院分区：
文献类型：
--
作者：
Ikawa, Y;Tsuda, K;Inoue, T
在原子水平上对分子识别的任意操纵有许多应用。然而，基于这种操作的人工酶的系统设计和从头合成一直是化学和生物技术领域的长期挑战。在这份报告中，我们通过实施一种合成策略开发了一种人工RNA连接酶，该策略将一系列基于天然存在的RNA-RNA识别基序的3D分子建模与模块化催化单元的小规模组合合成相融合。所得连接酶以模板导向的方式在反应位点处对两个核苷酸的任何组合产生3 '-5'键。反应速率是未催化反应的10（6）倍，产率高于以前报道的连接酶核酶。该策略可能适用于合成和开发各种非天然的功能性RNA与定义的三维结构。
Arbitrary manipulation of molecular recognition at the atomic level has many applications. However, systematic design and de novo synthesis of an artificial enzyme based on such manipulation has been a long-standing challenge in the field of chemistry and biotechnology. In this report, we developed an artificial RNA ligase by implementing a synthetic strategy that fuses a series of 3D molecular modelings based on naturally occurring RNA-RNA recognition motifs with a small-scale combinatorial synthesis of a modular catalytic unit. The resulting ligase produces a 3'-5' linkage in a template-directed manner for any combinations of two nucleotides at the reaction site. The reaction rate is 10(6)-fold over that of the uncatalyzed reaction with a yield higher than those of previously reported ligase ribozymes. The strategy may be applicable to the synthesis and development of a variety of nonnatural functional-RNAs with defined 3D structures.