合成一种原子特异性 H/C 标记的尿嘧啶核糖核苷磷酸酰胺。

Synthesis of an Atom-Specifically H/ C-Labeled Uridine Ribonucleoside Phosphoramidite.

机构信息

Center for Biomolecular Structure and Organization, Department of Chemistry and Biochemistry, University of Maryland, College Park, Maryland.

Current Address: Institute for Bioscience and Biotechnology Research, National Institute of Standards and Technology and the University of Maryland, Rockville, Maryland.

出版信息

Curr Protoc. 2022 Jul;2(7):e481. doi: 10.1002/cpz1.481.

DOI:10.1002/cpz1.481

PMID:35862131

Abstract

A combined enzymatic and chemical synthesis of a 2'-O-cyanoethoxymethyl (CEM) protected [1',6- C , 5- H]-uridine phosphoramidite is described herein. This is the first report of an atom-specific nucleobase and ribose labeled 2'-O-CEM protected ribonucleoside phosphoramidite. Importantly, the CEM 2'-OH protecting group permits the efficient solid-phase synthesis of large (>60 nucleotides) RNAs with good yield and purity. The new isotope-labeled phosphoramidite can therefore be applied to nuclear magnetic resonance (NMR) spectroscopy studies. Specifically, the [1',6- C , 5- H]-uridine phosphoramidite can be used to make position-specifically labeled RNAs for NMR analysis without complications from resonance overlap and scalar and dipolar couplings. © 2022 Wiley Periodicals LLC. Basic Protocol 1: Synthesis of the ribonucleoside 6 Basic Protocol 2: Synthesis of the ribonucleoside phosphoramidite 11.

摘要

本文描述了一种 2'-O-氰乙氧基甲基（CEM）保护的[1',6-C,5-H]-尿苷磷酸酰胺的酶促和化学联合合成。这是首例原子特异性碱基和核糖标记的 2'-O-CEM 保护的核糖核苷磷酸酰胺的报道。重要的是，CEM 2'-OH 保护基允许高效地固相合成具有良好收率和纯度的大（>60 个核苷酸）RNA。因此，这种新的同位素标记的磷酰胺可以应用于核磁共振（NMR）光谱研究。具体来说，[1',6-C,5-H]-尿苷磷酸酰胺可用于制备用于 NMR 分析的位置特异性标记的 RNA，而不会因共振重叠以及标量和偶极耦合而产生干扰。© 2022 Wiley Periodicals LLC. 基础方案 1：核糖核苷的合成 6 基础方案 2：核糖核苷磷酸酰胺的合成 11.