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Isotope Labels Combined with Solution NMR Spectroscopy Make Visible the Invisible Conformations of Small-to-Large RNAs
Chemical Reviews ( IF 62.1 ) Pub Date : 2022-04-20 , DOI: 10.1021/acs.chemrev.1c00845
Theodore K Dayie 1 , Lukasz T Olenginski 1 , Kehinde M Taiwo 1
Affiliation  

RNA is central to the proper function of cellular processes important for life on earth and implicated in various medical dysfunctions. Yet, RNA structural biology lags significantly behind that of proteins, limiting mechanistic understanding of RNA chemical biology. Fortunately, solution NMR spectroscopy can probe the structural dynamics of RNA in solution at atomic resolution, opening the door to their functional understanding. However, NMR analysis of RNA, with only four unique ribonucleotide building blocks, suffers from spectral crowding and broad linewidths, especially as RNAs grow in size. One effective strategy to overcome these challenges is to introduce NMR-active stable isotopes into RNA. However, traditional uniform labeling methods introduce scalar and dipolar couplings that complicate the implementation and analysis of NMR measurements. This challenge can be circumvented with selective isotope labeling. In this review, we outline the development of labeling technologies and their application to study biologically relevant RNAs and their complexes ranging in size from 5 to 300 kDa by NMR spectroscopy.

中文翻译:

同位素标记与溶液核磁共振波谱相结合,使小到大 RNA 的不可见构象变得可见

RNA对于地球生命重要的细胞过程的正常功能至关重要,并与各种医疗功能障碍有关。然而,RNA 结构生物学明显落后于蛋白质,限制了对 RNA 化学生物学的机制理解。幸运的是,溶液核磁共振波谱可以以原子分辨率探测溶液中 RNA 的结构动力学,为理解它们的功能打开了大门。然而,RNA 的 NMR 分析仅包含四种独特的核糖核苷酸构建模块,因此会遇到光谱拥挤和宽线宽的问题,尤其是当 RNA 尺寸增大时。克服这些挑战的一种有效策略是将 NMR 活性稳定同位素引入 RNA 中。然而,传统的统一标记方法引入了标量和偶极耦合,使 NMR 测量的实施和分析变得复杂。可以通过选择性同位素标记来规避这一挑战。在这篇综述中,我们概述了标记技术的发展及其应用,通过核磁共振波谱研究生物学相关的 RNA 及其复合物,其大小范围为 5 至 300 kDa。
更新日期:2022-04-20
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