通过结晶后处理提高 RNA 晶体衍射质量。
Improving RNA Crystal Diffraction Quality by Postcrystallization Treatment.
机构信息
Laboratory of Molecular Biology, National Institute of Diabetes and Digestive and Kidney Diseases, Bethesda, MD, USA.
Biochemistry and Biophysics Center, National Heart, Lung and Blood Institute, Bethesda, MD, USA.
出版信息
Methods Mol Biol. 2021;2323:25-37. doi: 10.1007/978-1-0716-1499-0_3.
The crystallization and structural determination of large RNAs and their complexes remain major bottlenecks in the mechanistic analysis of cellular and viral RNAs. Here, we describe a protocol that combines postcrystallization dehydration and ion replacement that dramatically improved the diffraction quality of crystals of a large gene-regulatory tRNA-mRNA complex. Through this method, the resolution limit of X-ray data extended from 8.5 to 3.2 Å, enabling structure determination. Although this protocol was developed for a particular RNA complex, the general importance of solvent and counterions in nucleic acid structure may render it generally useful for crystallographic analysis of other RNAs.
结晶和大 RNA 及其复合物的结构测定仍然是细胞和病毒 RNA 机械分析的主要瓶颈。在这里，我们描述了一种结合结晶后脱水和离子置换的方案，该方案显著提高了一个大型基因调控 tRNA-mRNA 复合物晶体的衍射质量。通过这种方法，X 射线数据的分辨率从 8.5 Å 提高到 3.2 Å，从而实现了结构测定。尽管该方案是针对特定的 RNA 复合物开发的，但溶剂和抗衡离子在核酸结构中的普遍重要性可能使其对其他 RNA 的晶体学分析普遍有用。
Zotero 插件