Development of solid state NMR methodology to study RNA and protein-RNA complexes

在这个研究项目中，我们计划开发新的核磁共振方法，通过固体核磁共振(ss核磁共振)来研究RNA和RNA-蛋白质复合体的结构。单核磁共振利用了中等尺寸和高溶解度的要求，这限制了通过溶液状态核磁共振进行结构研究，或者需要高分辨率的衍射晶体来通过X射线晶体学来确定结构。单链核磁共振在淀粉样纤维、膜蛋白和大型蛋白质复合体等领域发展迅速，但由于缺乏一种通用的单链核磁共振研究RNA的方法，对RNA或蛋白质-RNA复合体(RNPs)的研究较少。利用核磁共振硬件的最新发展、不同的标记技术和现代选择性磁化转移方案，我们将开发一种通过单核磁共振确定RNA结构的策略。这些结果将为研究大的RNA和RNPs铺平道路，这些RNAs和RNPs既不能用结晶学也不能用溶液状态核磁共振研究。

In this research project we plan to develop novel NMR methodology for structural studies of RNA and RNA-protein complexes by means of solid-state NMR (ssNMR). ssNMR leverages the requirements of moderate size and high solubility that limit structural studies by solution-state NMR, or the necessity of high-resolution diffracting crystals for structure determination by X-ray crystallography. ssNMR has been developing fast in the fields of amyloid fibrils, membrane proteins and large protein complexes; however, significantly fewer studies have been performed on RNA or protein-RNA complexes (RNPs), due to the lack of a general approach to study RNA by ssNMR. Taking advantage of recent development of the NMR hardware, different labeling techniques and modern selective magnetization transfer schemes, we will develop a strategy to determine the structure of RNA by ssNMR. These results will pave the way to study large RNAs and RNPs that cannot be studied either by crystallography or solution-state NMR.