Institute for Organic Chemistry and Centre of Biomolecular Drug Research (BMWZ), Leibniz University Hannover, Schneiderberg 38, 30167, Hannover, Germany.
Group of NMR-based Structural Chemistry, Helmholtz Centre of Infection Research, Inhoffenstrasse 7, 38124, Braunschweig, Germany.
Angew Chem Int Ed Engl. 2021 Oct 25;60(44):23903-23910. doi: 10.1002/anie.202107263. Epub 2021 Sep 28.
Knowledge of RNA structure, either in isolation or in complex, is fundamental to understand the mechanism of cellular processes. Solid-state NMR (ssNMR) is applicable to high molecular-weight complexes and does not require crystallization; thus, it is well-suited to study RNA as part of large multicomponent assemblies. Recently, we solved the first structures of both RNA and an RNA-protein complex by ssNMR using conventional C- and N-detection. This approach is limited by the severe overlap of the RNA peaks together with the low sensitivity of multidimensional experiments. Here, we overcome the limitations in sensitivity and resolution by using H-detection at fast MAS rates. We develop experiments that allow the identification of complete nucleobase spin-systems together with their site-specific base pair pattern using sub-milligram quantities of one uniformly labelled RNA sample. These experiments provide rapid access to RNA secondary structure by ssNMR in protein-RNA complexes of any size.
对 RNA 结构的了解，无论是单独的还是复杂的，都是理解细胞过程机制的基础。固态 NMR（ssNMR）适用于高分子量复合物，且不需要结晶；因此，它非常适合作为研究 RNA 的一部分，作为大型多组分组装体的一部分。最近，我们使用传统的 C 和 N 检测，通过 ssNMR 首次解决了 RNA 及其 RNA-蛋白质复合物的结构问题。这种方法受到 RNA 峰严重重叠以及多维实验灵敏度低的限制。在这里，我们通过在快速 MAS 速率下使用 H 检测克服了灵敏度和分辨率的限制。我们开发了实验，使用亚毫克量的一种均一标记的 RNA 样品，即可识别完整的核碱基自旋系统及其特定于位置的碱基对模式。这些实验为任何大小的蛋白质-RNA 复合物中的 ssNMR 提供了快速获取 RNA 二级结构的途径。
