Open AccessCrystallization of RNA polymerase I subcomplex A14/A43 by iterative prediction, probing and removal of flexible regions
Author(s) -
Geiger Sebastian R.,
Kuhn Claus.D.,
Leidig Christoph,
Renkawitz Jörg,
Cramer Patrick
Publication year - 2008
Publication title -
acta crystallographica section f
Language(s) - English
Resource type - Journals
ISSN - 1744-3091
DOI - 10.1107/s174430910800972x
Subject(s) - crystallization , rna , polymerase , macromolecule , rna polymerase , chemistry , computational biology , biophysics , crystallography , biology , biochemistry , dna , gene , organic chemistry
The removal of flexible protein regions is generally used to promote crystallization, but advanced strategies to quickly remove multiple flexible regions from proteins or protein complexes are lacking. Here, it is shown how a protein heterodimer with multiple flexibilities, the RNA polymerase I subcomplex A14/A43, could be crystallized with the use of an iterative procedure of predicting flexible regions, experimentally testing and improving these predictions and combining deletions of flexible regions in a stepwise manner. This strategy should enable the crystallization of other proteins and subcomplexes with multiple flexibilities, as required for hybrid structure solution of large macromolecular assemblies.