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Iterative screen optimization maximizes the efficiency of macromolecular crystallization

Advances in X-ray crystallography have streamlined the process of determining high-resolution three-dimensional macromolecular structures. However, a rate-limiting step in this process continues to be the generation of crystals that are of sufficient size and quality for subsequent diffraction exper...

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Detalles Bibliográficos
Autores principales: Jones, Harrison G., Wrapp, Daniel, Gilman, Morgan S. A., Battles, Michael B., Wang, Nianshuang, Sacerdote, Sofia, Chuang, Gwo-Yu, Kwong, Peter D., McLellan, Jason S.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: International Union of Crystallography 2019
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6360444/
https://www.ncbi.nlm.nih.gov/pubmed/30713164
http://dx.doi.org/10.1107/S2053230X18017338
Descripción
Sumario:Advances in X-ray crystallography have streamlined the process of determining high-resolution three-dimensional macromolecular structures. However, a rate-limiting step in this process continues to be the generation of crystals that are of sufficient size and quality for subsequent diffraction experiments. Here, iterative screen optimization (ISO), a highly automated process in which the precipitant concentrations of each condition in a crystallization screen are modified based on the results of a prior crystallization experiment, is described. After designing a novel high-throughput crystallization screen to take full advantage of this method, the value of ISO is demonstrated by using it to successfully crystallize a panel of six diverse proteins. The results suggest that ISO is an effective method to obtain macromolecular crystals, particularly for proteins that crystallize under a narrow range of precipitant concentrations.