Structural and biochemical characterization of DSL ribozyme
Structural and biochemical characterization of DSL ribozyme
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DOI：
10.1016/j.bbrc.2005.11.007
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发表时间：
2006-01-06
影响因子：
3.1
通讯作者：
Inoue, T
中科院分区：
文献类型：
--
作者：
Horie, S;Ikawa, Y;Inoue, T
我们最近报道了一种新的RNA连接酶核酶（DSL）的分子设计与合成，它的活性位点是从由30个随机核苷酸组成的序列库中选择的，它定义了它所设计的RNA支架的三维结构。本文报道了DSL的结构和生化分析.结构分析表明，活性位点，其中包括选定的序列，连接到折叠支架的设计。为了观察DSL是否类似于已知的核酶，进行了生物化学测定。金属依赖的动力学研究表明，连接酶需要镁离子。用Co（NH3）（6）（3+）取代Mg 2+抑制了反应，表明DSL需要Mg 2+的内球配位进行连接反应。结果表明，DSL具有类似于先前报道的催化RNA的要求。(c)2005年爱思唯尔公司All rights reserved.
We recently reported oil the molecular design and synthesis of a new RNA ligase ribozyme (DSL), whose active site was selected front a sequence library consisting of 30 random nucleotides set oil it defined 3D structure of it designed RNA scaffold. In this study, we report Oil the structural and biochemical analyses of DSL. Structural analysis indicates that the active site, which consists of the selected sequence, attaches to the folded scaffold as designed. To see whether DSL resembles known ribozymes, a biochemical assay was performed. Metal-dependent kinetic studies suggest that the ligase requires Mg2+ ions. The replacement of Mg2+ with Co(NH3)(6)(3+) prohibits the reaction, indicating that DSL requires innersphere coordination of Mg2+ for a ligation reaction. The results show that DSL has requirements similar to those of previously reported catalytic RNAs. (c) 2005 Elsevier Inc. All rights reserved.