Cargando…
Defined PEG smears as an alternative approach to enhance the search for crystallization conditions and crystal-quality improvement in reduced screens
The quest for an optimal limited set of effective crystallization conditions remains a challenge in macromolecular crystallography, an issue that is complicated by the large number of chemicals which have been deemed to be suitable for promoting crystal growth. The lack of rational approaches toward...
Autores principales: | , , |
---|---|
Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
International Union of Crystallography
2015
|
Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4528798/ https://www.ncbi.nlm.nih.gov/pubmed/26249344 http://dx.doi.org/10.1107/S1399004715007968 |
_version_ | 1782384707667755008 |
---|---|
author | Chaikuad, Apirat Knapp, Stefan von Delft, Frank |
author_facet | Chaikuad, Apirat Knapp, Stefan von Delft, Frank |
author_sort | Chaikuad, Apirat |
collection | PubMed |
description | The quest for an optimal limited set of effective crystallization conditions remains a challenge in macromolecular crystallography, an issue that is complicated by the large number of chemicals which have been deemed to be suitable for promoting crystal growth. The lack of rational approaches towards the selection of successful chemical space and representative combinations has led to significant overlapping conditions, which are currently present in a multitude of commercially available crystallization screens. Here, an alternative approach to the sampling of widely used PEG precipitants is suggested through the use of PEG smears, which are mixtures of different PEGs with a requirement of either neutral or cooperatively positive effects of each component on crystal growth. Four newly defined smears were classified by molecular-weight groups and enabled the preservation of specific properties related to different polymer sizes. These smears not only allowed a wide coverage of properties of these polymers, but also reduced PEG variables, enabling greater sampling of other parameters such as buffers and additives. The efficiency of the smear-based screens was evaluated on more than 220 diverse recombinant human proteins, which overall revealed a good initial crystallization success rate of nearly 50%. In addition, in several cases successful crystallizations were only obtained using PEG smears, while various commercial screens failed to yield crystals. The defined smears therefore offer an alternative approach towards PEG sampling, which will benefit the design of crystallization screens sampling a wide chemical space of this key precipitant. |
format | Online Article Text |
id | pubmed-4528798 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2015 |
publisher | International Union of Crystallography |
record_format | MEDLINE/PubMed |
spelling | pubmed-45287982015-08-24 Defined PEG smears as an alternative approach to enhance the search for crystallization conditions and crystal-quality improvement in reduced screens Chaikuad, Apirat Knapp, Stefan von Delft, Frank Acta Crystallogr D Biol Crystallogr Research Papers The quest for an optimal limited set of effective crystallization conditions remains a challenge in macromolecular crystallography, an issue that is complicated by the large number of chemicals which have been deemed to be suitable for promoting crystal growth. The lack of rational approaches towards the selection of successful chemical space and representative combinations has led to significant overlapping conditions, which are currently present in a multitude of commercially available crystallization screens. Here, an alternative approach to the sampling of widely used PEG precipitants is suggested through the use of PEG smears, which are mixtures of different PEGs with a requirement of either neutral or cooperatively positive effects of each component on crystal growth. Four newly defined smears were classified by molecular-weight groups and enabled the preservation of specific properties related to different polymer sizes. These smears not only allowed a wide coverage of properties of these polymers, but also reduced PEG variables, enabling greater sampling of other parameters such as buffers and additives. The efficiency of the smear-based screens was evaluated on more than 220 diverse recombinant human proteins, which overall revealed a good initial crystallization success rate of nearly 50%. In addition, in several cases successful crystallizations were only obtained using PEG smears, while various commercial screens failed to yield crystals. The defined smears therefore offer an alternative approach towards PEG sampling, which will benefit the design of crystallization screens sampling a wide chemical space of this key precipitant. International Union of Crystallography 2015-07-28 /pmc/articles/PMC4528798/ /pubmed/26249344 http://dx.doi.org/10.1107/S1399004715007968 Text en © Chaikuad et al. 2015 http://creativecommons.org/licenses/by/2.0/uk/ This is an open-access article distributed under the terms of the Creative Commons Attribution Licence, which permits unrestricted use, distribution, and reproduction in any medium, provided the original authors and source are cited. |
spellingShingle | Research Papers Chaikuad, Apirat Knapp, Stefan von Delft, Frank Defined PEG smears as an alternative approach to enhance the search for crystallization conditions and crystal-quality improvement in reduced screens |
title | Defined PEG smears as an alternative approach to enhance the search for crystallization conditions and crystal-quality improvement in reduced screens |
title_full | Defined PEG smears as an alternative approach to enhance the search for crystallization conditions and crystal-quality improvement in reduced screens |
title_fullStr | Defined PEG smears as an alternative approach to enhance the search for crystallization conditions and crystal-quality improvement in reduced screens |
title_full_unstemmed | Defined PEG smears as an alternative approach to enhance the search for crystallization conditions and crystal-quality improvement in reduced screens |
title_short | Defined PEG smears as an alternative approach to enhance the search for crystallization conditions and crystal-quality improvement in reduced screens |
title_sort | defined peg smears as an alternative approach to enhance the search for crystallization conditions and crystal-quality improvement in reduced screens |
topic | Research Papers |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4528798/ https://www.ncbi.nlm.nih.gov/pubmed/26249344 http://dx.doi.org/10.1107/S1399004715007968 |
work_keys_str_mv | AT chaikuadapirat definedpegsmearsasanalternativeapproachtoenhancethesearchforcrystallizationconditionsandcrystalqualityimprovementinreducedscreens AT knappstefan definedpegsmearsasanalternativeapproachtoenhancethesearchforcrystallizationconditionsandcrystalqualityimprovementinreducedscreens AT vondelftfrank definedpegsmearsasanalternativeapproachtoenhancethesearchforcrystallizationconditionsandcrystalqualityimprovementinreducedscreens |