Dramatic Improvement of Crystals of Large RNAs by Cation Replacement and Dehydration

  • Zhang, Jinwei
  • Ferré-D’Amaré, Adrian R.
Publication date
September 2014
Publisher
Elsevier Ltd.

Abstract

SummaryCompared to globular proteins, RNAs with complex 3D folds are characterized by poorly differentiated molecular surfaces dominated by backbone phosphates, sparse tertiary contacts stabilizing global architecture, and conformational flexibility. The resulting generally poor order of crystals of large RNAs and their complexes frequently hampers crystallographic structure determination. We describe and rationalize a postcrystallization treatment strategy that exploits the importance of solvation and counterions for RNA folding. Replacement of Li+ and Mg2+ needed for growth of crystals of a tRNA-riboswitch-protein complex with Sr2+, coupled with dehydration, dramatically improved the resolution limit (8.5–3.2 Å) and data quality, enabling...

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