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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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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
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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.
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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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