Synthesis of atom-specific nucleobase and ribose labeled uridine phosphoramidite for NMR analysis of large RNAs
Synthesis of atom-specific nucleobase and ribose labeled uridine phosphoramidite for NMR analysis of large RNAs
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DOI：
10.1007/s00706-021-02851-2
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发表时间：
2021-10-18
影响因子：
1.8
通讯作者：
Dayie, Theodore K.
中科院分区：
文献类型：
--
作者：
Olenginski, Lukasz T.;Becette, Owen B.;Dayie, Theodore K.
我们描述了一种2'- o -氰乙氧基甲基（CEM） [1',6-C-13(2)， 5-H-2]-尿苷磷酰胺（amidite）的混合酶和化学合成。这是首次报道一种原子特异性核碱基和核糖标记的CEM酰胺。重要的是，CEM 2'-OH保护基团允许高效的固相合成大（bbb60核苷酸）rna，具有良好的产量和纯度。因此，我们的同位素标记的中间体可以用来制作大的、位置特异性标记的rna，用于核磁共振分析，而不会出现共振重叠、标量和偶极耦合的并发症。
We describe the hybrid enzymatic and chemical synthesis of a 2'-O-cyanoethoxymethyl (CEM) [1',6-C-13(2), 5-H-2]-uridine phosphoramidite (amidite). This is the first report of an atom-specific nucleobase and ribose labeled CEM amidite. Importantly, the CEM 2'-OH protecting group permits the efficient solid-phase synthesis of large (> 60 nucleotides) RNAs with good yield and purity. Therefore, our isotope-labeled amidite can be used to make large, position-specifically labeled RNAs for NMR analysis without complications from resonance overlap and scalar and dipolar couplings.[GRAPHICS].