Post-crystallization Improvement of RNA Crystal Diffraction Quality.
Publication Type:Journal Article
Source:Methods Mol Biol, Volume 1316, p.13-24 (2015)
Keywords:Crystallization, Nucleic Acid Conformation, RNA, RNA, Ribosomal, RNA, Transfer
<p>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 post-crystallization 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.</p>