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Protein-to-structure pipeline for ambient-temperature in situ crystallography at VMXi
The utility of X-ray crystal structures determined under ambient-temperature conditions is becoming increasingly recognized. Such experiments can allow protein dynamics to be characterized and are particularly well suited to challenging protein targets that may form fragile crystals that are difficu...
Autores principales: | , , , , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
International Union of Crystallography
2023
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10324489/ https://www.ncbi.nlm.nih.gov/pubmed/37199504 http://dx.doi.org/10.1107/S2052252523003810 |
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author | Mikolajek, Halina Sanchez-Weatherby, Juan Sandy, James Gildea, Richard J. Campeotto, Ivan Cheruvara, Harish Clarke, John D. Foster, Toshana Fujii, Sotaro Paulsen, Ian T. Shah, Bhumika S. Hough, Michael A. |
author_facet | Mikolajek, Halina Sanchez-Weatherby, Juan Sandy, James Gildea, Richard J. Campeotto, Ivan Cheruvara, Harish Clarke, John D. Foster, Toshana Fujii, Sotaro Paulsen, Ian T. Shah, Bhumika S. Hough, Michael A. |
author_sort | Mikolajek, Halina |
collection | PubMed |
description | The utility of X-ray crystal structures determined under ambient-temperature conditions is becoming increasingly recognized. Such experiments can allow protein dynamics to be characterized and are particularly well suited to challenging protein targets that may form fragile crystals that are difficult to cryo-cool. Room-temperature data collection also enables time-resolved experiments. In contrast to the high-throughput highly automated pipelines for determination of structures at cryogenic temperatures widely available at synchrotron beamlines, room-temperature methodology is less mature. Here, the current status of the fully automated ambient-temperature beamline VMXi at Diamond Light Source is described, and a highly efficient pipeline from protein sample to final multi-crystal data analysis and structure determination is shown. The capability of the pipeline is illustrated using a range of user case studies representing different challenges, and from high and lower symmetry space groups and varied crystal sizes. It is also demonstrated that very rapid structure determination from crystals in situ within crystallization plates is now routine with minimal user intervention. |
format | Online Article Text |
id | pubmed-10324489 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2023 |
publisher | International Union of Crystallography |
record_format | MEDLINE/PubMed |
spelling | pubmed-103244892023-07-07 Protein-to-structure pipeline for ambient-temperature in situ crystallography at VMXi Mikolajek, Halina Sanchez-Weatherby, Juan Sandy, James Gildea, Richard J. Campeotto, Ivan Cheruvara, Harish Clarke, John D. Foster, Toshana Fujii, Sotaro Paulsen, Ian T. Shah, Bhumika S. Hough, Michael A. IUCrJ Research Papers The utility of X-ray crystal structures determined under ambient-temperature conditions is becoming increasingly recognized. Such experiments can allow protein dynamics to be characterized and are particularly well suited to challenging protein targets that may form fragile crystals that are difficult to cryo-cool. Room-temperature data collection also enables time-resolved experiments. In contrast to the high-throughput highly automated pipelines for determination of structures at cryogenic temperatures widely available at synchrotron beamlines, room-temperature methodology is less mature. Here, the current status of the fully automated ambient-temperature beamline VMXi at Diamond Light Source is described, and a highly efficient pipeline from protein sample to final multi-crystal data analysis and structure determination is shown. The capability of the pipeline is illustrated using a range of user case studies representing different challenges, and from high and lower symmetry space groups and varied crystal sizes. It is also demonstrated that very rapid structure determination from crystals in situ within crystallization plates is now routine with minimal user intervention. International Union of Crystallography 2023-05-19 /pmc/articles/PMC10324489/ /pubmed/37199504 http://dx.doi.org/10.1107/S2052252523003810 Text en © Halina Mikolajek et al. 2023 https://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. |
spellingShingle | Research Papers Mikolajek, Halina Sanchez-Weatherby, Juan Sandy, James Gildea, Richard J. Campeotto, Ivan Cheruvara, Harish Clarke, John D. Foster, Toshana Fujii, Sotaro Paulsen, Ian T. Shah, Bhumika S. Hough, Michael A. Protein-to-structure pipeline for ambient-temperature in situ crystallography at VMXi |
title | Protein-to-structure pipeline for ambient-temperature in situ crystallography at VMXi |
title_full | Protein-to-structure pipeline for ambient-temperature in situ crystallography at VMXi |
title_fullStr | Protein-to-structure pipeline for ambient-temperature in situ crystallography at VMXi |
title_full_unstemmed | Protein-to-structure pipeline for ambient-temperature in situ crystallography at VMXi |
title_short | Protein-to-structure pipeline for ambient-temperature in situ crystallography at VMXi |
title_sort | protein-to-structure pipeline for ambient-temperature in situ crystallography at vmxi |
topic | Research Papers |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10324489/ https://www.ncbi.nlm.nih.gov/pubmed/37199504 http://dx.doi.org/10.1107/S2052252523003810 |
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