Post-crystallization Improvement of RNA Crystals by Synergistic Ion Exchange and Dehydration.

ABSTRACT: Compared to the recent dramatic growth in the numbers of genome-wide and functional studies of complex non-coding RNAs, mechanistic and structural analyses have lagged behind. A major technical bottleneck in the structural determination of large RNAs and their complexes is preparation of diffracting crystals. Empirically, a vast majority of such RNA crystals fail to diffract X-rays to usable resolution (~4 Å) due to their inherent disorder and non-specific packing within the crystals. Here, we present a protocol that combines post-crystallization cation replacement and dehydration that dramatically improved the diffraction quality of crystals of a large gene-regulatory mRNA-tRNA complex. This procedure not only extended the resolution limit of X-ray data from 8.5 to 3.2 Å, but also signifi

SUBMITTER: Zhang J 

PROVIDER: S-EPMC4943581 | biostudies-literature | 2015

REPOSITORIES: biostudies-literature