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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...
Autores principales: | , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
International Union of Crystallography
2019
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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 |
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author | 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. |
author_facet | 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. |
author_sort | Jones, Harrison G. |
collection | PubMed |
description | 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. |
format | Online Article Text |
id | pubmed-6360444 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2019 |
publisher | International Union of Crystallography |
record_format | MEDLINE/PubMed |
spelling | pubmed-63604442019-02-11 Iterative screen optimization maximizes the efficiency of macromolecular crystallization 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. Acta Crystallogr F Struct Biol Commun Research Communications 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. International Union of Crystallography 2019-01-24 /pmc/articles/PMC6360444/ /pubmed/30713164 http://dx.doi.org/10.1107/S2053230X18017338 Text en © Jones et al. 2019 http://creativecommons.org/licenses/by/4.0/ This is an open-access article distributed under the terms of the Creative Commons Attribution (CC-BY) Licence, which permits unrestricted use, distribution, and reproduction in any medium, provided the original authors and source are cited.http://creativecommons.org/licenses/by/4.0/ |
spellingShingle | Research Communications 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. Iterative screen optimization maximizes the efficiency of macromolecular crystallization |
title | Iterative screen optimization maximizes the efficiency of macromolecular crystallization |
title_full | Iterative screen optimization maximizes the efficiency of macromolecular crystallization |
title_fullStr | Iterative screen optimization maximizes the efficiency of macromolecular crystallization |
title_full_unstemmed | Iterative screen optimization maximizes the efficiency of macromolecular crystallization |
title_short | Iterative screen optimization maximizes the efficiency of macromolecular crystallization |
title_sort | iterative screen optimization maximizes the efficiency of macromolecular crystallization |
topic | Research Communications |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6360444/ https://www.ncbi.nlm.nih.gov/pubmed/30713164 http://dx.doi.org/10.1107/S2053230X18017338 |
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