Chemo-enzymatic synthesis of [2-13C, 7-15 N]-ATP for facile NMR analysis of RNA
Chemo-enzymatic synthesis of [2-13C, 7-15 N]-ATP for facile NMR analysis of RNA
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[2-13C, 7-15N]-ATP 的化学酶合成用于 RNA 的简便 NMR 分析
DOI：
10.1007/s00706-020-02667-6
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发表时间：
2020
期刊：
影响因子：
--
通讯作者：
Dayie, Theodore K.
中科院分区：
文献类型：
--
作者：
Olenginski, Lukasz T.;Dayie, Theodore K.
摘要我们报道了一种有效的化学-酶促合成[2- 13 C，7- 15 N]-腺苷5′-三磷酸结构单元，用于通过基于T7 RNA聚合酶的体外转录掺入RNA。这种原子特异性标记消除了光谱拥挤，简化了溶液NMR光谱法对大RNA的分析。应用程序探针腺苷C2和N7网站在一个大的61个核苷酸的病毒RNA与二维和三维NMR实验证明。图形摘要
AbstractWe report the efficient chemo-enzymatic synthesis of an [2-13C, 7-15N]-adenosine 5′-triphosphate building block for incorporation into RNA via T7 RNA polymerase-based in vitro transcription. Such atom-specific labeling alleviates spectral crowding and simplifies the analysis of large RNAs by solution NMR spectroscopy. Applications to probe adenosine C2 and N7 sites in a large 61 nucleotide viral RNA with two- and three-dimensional NMR experiments are demonstrated.Graphic abstract