Chapter title |
Post-crystallization Improvement of RNA Crystal Diffraction Quality
|
---|---|
Chapter number | 2 |
Book title |
RNA Scaffolds
|
Published in |
Methods in molecular biology, January 2015
|
DOI | 10.1007/978-1-4939-2730-2_2 |
Pubmed ID | |
Book ISBNs |
978-1-4939-2729-6, 978-1-4939-2730-2
|
Authors |
Jinwei Zhang, Adrian R. Ferré-D’Amaré, Zhang, Jinwei, Ferré-D’Amaré, Adrian R. |
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. |
Mendeley readers
Geographical breakdown
Country | Count | As % |
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Unknown | 3 | 100% |
Demographic breakdown
Readers by professional status | Count | As % |
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Student > Ph. D. Student | 2 | 67% |
Other | 1 | 33% |
Readers by discipline | Count | As % |
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Biochemistry, Genetics and Molecular Biology | 1 | 33% |
Materials Science | 1 | 33% |
Unknown | 1 | 33% |