title: Post-crystallization Improvement of RNA Crystal Diffraction Quality.
abstract: 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.
authors.affiliation: National Heart, Lung and Blood Institute, 50 South Drive, MSC 8012, Bethesda, MD, 20892-8012, USA.
authors.forename: Jinwei
authors.initials: J
authors.lastname: Zhang
authors.forename: Adrian R
authors.initials: AR
authors.lastname: Ferré-D'Amaré
mesh_terms: D003460:Crystallization; D009690:Nucleic Acid Conformation; D012313:RNA / Q000737:chemistry*; D012335:RNA, Ribosomal / Q000737:chemistry; D012343:RNA, Transfer / Q000737:chemistry
chemical_list: D012335:RNA, Ribosomal; D012313:RNA; D012343:RNA, Transfer