Swiss Federal Institute of Technology Zürich, Zürich, Switzerland.
Adv Exp Med Biol. 2012;992:121-44. doi: 10.1007/978-94-007-4954-2_7.
NMR spectroscopy has become substantial in the elucidation of RNA structures and their complexes with other nucleic acids, proteins or small molecules. Almost half of the RNA structures deposited in the Protein Data Bank were determined by NMR spectroscopy, whereas NMR accounts for only 11% for proteins. Recent improvements in isotope labeling of RNA have strongly contributed to the high impact of NMR in RNA structure determination. In this book chapter, we review the advances in isotope labeling of RNA focusing on larger RNAs. We start by discussing several ways for the production and purification of large quantities of pure isotope labeled RNA. We continue by reviewing different strategies for selective deuteration of nucleotides. Finally, we present a comparison of several approaches for segmental isotope labeling of RNA. Selective deuteration of nucleotides in combination with segmental isotope labeling is paving the path for studying RNAs of ever increasing size.
NMR 光谱学在阐明 RNA 结构及其与其他核酸、蛋白质或小分子的复合物方面发挥了重要作用。几乎一半储存在蛋白质数据库中的 RNA 结构都是通过 NMR 光谱学确定的，而 NMR 方法在蛋白质中的占比仅为 11%。RNA 同位素标记的最新进展极大地促进了 NMR 在 RNA 结构测定中的高影响力。在本章中，我们将综述 RNA 同位素标记的最新进展，重点关注较大的 RNA。我们首先讨论了几种大量生产和纯化纯同位素标记 RNA 的方法。然后，我们回顾了核苷酸选择性氘代的不同策略。最后，我们比较了几种 RNA 分段同位素标记的方法。核苷酸的选择性氘代与分段同位素标记相结合，为研究越来越大的 RNA 铺平了道路。
