title: High-throughput crystallization screening.
abstract: Protein structure determination by X-ray crystallography is dependent on obtaining a single protein crystal suitable for diffraction data collection. Due to this requirement, protein crystallization represents a key step in protein structure determination. The conditions for protein crystallization have to be determined empirically for each protein, making this step also a bottleneck in the structure determination process. Typical protein crystallization practice involves parallel setup and monitoring of a considerable number of individual protein crystallization experiments (also called crystallization trials). In these trials the aliquots of purified protein are mixed with a range of solutions composed of a precipitating agent, buffer, and sometimes an additive that have been previously successful in prompting protein crystallization. The individual chemical conditions in which a particular protein shows signs of crystallization are used as a starting point for further crystallization experiments. The goal is optimizing the formation of individual protein crystals of sufficient size and quality to make them suitable for diffraction data collection. Thus the composition of the primary crystallization screen is critical for successful crystallization.Systematic analysis of crystallization experiments carried out on several hundred proteins as part of large-scale structural genomics efforts allowed the optimization of the protein crystallization protocol and identification of a minimal set of 96 crystallization solutions (the "TRAP" screen) that, in our experience, led to crystallization of the maximum number of proteins.
authors.affiliation: Department of Chemical Engineering and Applied Chemistry, University of Toronto, 200 College Street, Toronto, ON, Canada.
authors.forename: Tatiana
authors.initials: T
authors.lastname: Skarina
authors.forename: Xiaohui
authors.initials: X
authors.lastname: Xu
authors.forename: Elena
authors.initials: E
authors.lastname: Evdokimova
authors.forename: Alexei
authors.initials: A
authors.lastname: Savchenko
mesh_terms: D003460:Crystallization / Q000379:methods*; D018360:Crystallography, X-Ray; D057166:High-Throughput Screening Assays; D006801:Humans; D008967:Molecular Biology / Q000379:methods*; D011506:Proteins / Q000737:chemistry* / Q000302:isolation & purification
chemical_list: D011506:Proteins