Strategies for crystallization and structure determination of very large macromolecular assemblies.
High-resolution structures of macromolecular assemblies are pivotal for our understanding of their biological functions in fundamental cellular processes. In the field of X-ray crystallography, recent methodological and instrumental advances have led to the structure determinations of macromolecular assemblies of increased size and complexity, such as those of ribosomal complexes, RNA polymerases, and large multifunctional enzymes. These advances include the use of robotic screening techniques that maximize the chances of obtaining well-diffracting crystals of large complexes through the fine sampling of crystallization space. Sophisticated crystal optimization and cryoprotection techniques and the use of highly brilliant X-ray beams from third-generation synchrotron light sources now allow data collection from weakly diffracting crystals with large asymmetric units. Combined approaches are used to derive phase information, including phases calculated from electron microscopy (EM) models, heavy atom clusters, and density modification protocols. New crystallographic software tools prove valuable for structure determination and model refinement of large macromolecular complexes.
Mueller
Marcus
M
Institute of Molecular Biology and Biophysics, ETH Zurich, 8093 Zurich, Switzerland.
Jenni
Simon
S
Ban
Nenad
N
D003460:Crystallization / Q000379:methods*; D018360:Crystallography, X-Ray; D054889:Fatty Acid Synthase, Type II / Q000737:chemistry / Q000302:isolation & purification; D046911:Macromolecular Substances / Q000737:chemistry* / Q000302:isolation & purification*; D008958:Models, Molecular; D015394:Molecular Structure; D009113:Muramidase / Q000737:chemistry / Q000302:isolation & purification; D012319:RNA Polymerase II / Q000737:chemistry / Q000302:isolation & purification; D055672:Static Electricity
D046911:Macromolecular Substances; D012319:RNA Polymerase II; D009113:Muramidase; D054889:Fatty Acid Synthase, Type II