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.
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Authors | Jinwei Zhang, Adrian R Ferré-D'Amaré |
Journal | Methods in molecular biology (Clifton, N.J.)
(Methods Mol Biol)
Vol. 1316
Pg. 13-24
( 2015)
ISSN: 1940-6029 [Electronic] United States |
PMID | 25967049
(Publication Type: Journal Article, Research Support, N.I.H., Intramural, Research Support, U.S. Gov't, Non-P.H.S.)
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Chemical References |
- RNA, Ribosomal
- RNA
- RNA, Transfer
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Topics |
- Crystallization
- Nucleic Acid Conformation
- RNA
(chemistry)
- RNA, Ribosomal
(chemistry)
- RNA, Transfer
(chemistry)
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