Improving RNA Crystal Diffraction Quality by Postcrystallization Treatment.
RNA SCAFFOLDS, 2 EDITION: Methods and Protocols(2021)
摘要
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.
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关键词
Crystal dehydration,Ion replacement,Riboswitch,T-box RNA,X-ray Crystallography,tRNA
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