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Perdeuteration, crystallization, data collection and comparison of five neutron diffraction data sets of complexes of human galectin-3C
Galectin-3 is an important protein in molecular signalling events involving carbohydrate recognition, and an understanding of the hydrogen-bonding patterns in the carbohydrate-binding site of its C-terminal domain (galectin-3C) is important for the development of new potent inhibitors. The authors a...
Autores principales: | , , , , , , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
International Union of Crystallography
2016
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5108347/ https://www.ncbi.nlm.nih.gov/pubmed/27841752 http://dx.doi.org/10.1107/S2059798316015540 |
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author | Manzoni, Francesco Saraboji, Kadhirvel Sprenger, Janina Kumar, Rohit Noresson, Ann-Louise Nilsson, Ulf J. Leffler, Hakon Fisher, S. Zoë Schrader, Tobias E. Ostermann, Andreas Coates, Leighton Blakeley, Matthew P. Oksanen, Esko Logan, Derek T. |
author_facet | Manzoni, Francesco Saraboji, Kadhirvel Sprenger, Janina Kumar, Rohit Noresson, Ann-Louise Nilsson, Ulf J. Leffler, Hakon Fisher, S. Zoë Schrader, Tobias E. Ostermann, Andreas Coates, Leighton Blakeley, Matthew P. Oksanen, Esko Logan, Derek T. |
author_sort | Manzoni, Francesco |
collection | PubMed |
description | Galectin-3 is an important protein in molecular signalling events involving carbohydrate recognition, and an understanding of the hydrogen-bonding patterns in the carbohydrate-binding site of its C-terminal domain (galectin-3C) is important for the development of new potent inhibitors. The authors are studying these patterns using neutron crystallography. Here, the production of perdeuterated human galectin-3C and successive improvement in crystal size by the development of a crystal-growth protocol involving feeding of the crystallization drops are described. The larger crystals resulted in improved data quality and reduced data-collection times. Furthermore, protocols for complete removal of the lactose that is necessary for the production of large crystals of apo galectin-3C suitable for neutron diffraction are described. Five data sets have been collected at three different neutron sources from galectin-3C crystals of various volumes. It was possible to merge two of these to generate an almost complete neutron data set for the galectin-3C–lactose complex. These data sets provide insights into the crystal volumes and data-collection times necessary for the same system at sources with different technologies and data-collection strategies, and these insights are applicable to other systems. |
format | Online Article Text |
id | pubmed-5108347 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2016 |
publisher | International Union of Crystallography |
record_format | MEDLINE/PubMed |
spelling | pubmed-51083472016-11-17 Perdeuteration, crystallization, data collection and comparison of five neutron diffraction data sets of complexes of human galectin-3C Manzoni, Francesco Saraboji, Kadhirvel Sprenger, Janina Kumar, Rohit Noresson, Ann-Louise Nilsson, Ulf J. Leffler, Hakon Fisher, S. Zoë Schrader, Tobias E. Ostermann, Andreas Coates, Leighton Blakeley, Matthew P. Oksanen, Esko Logan, Derek T. Acta Crystallogr D Struct Biol Research Papers Galectin-3 is an important protein in molecular signalling events involving carbohydrate recognition, and an understanding of the hydrogen-bonding patterns in the carbohydrate-binding site of its C-terminal domain (galectin-3C) is important for the development of new potent inhibitors. The authors are studying these patterns using neutron crystallography. Here, the production of perdeuterated human galectin-3C and successive improvement in crystal size by the development of a crystal-growth protocol involving feeding of the crystallization drops are described. The larger crystals resulted in improved data quality and reduced data-collection times. Furthermore, protocols for complete removal of the lactose that is necessary for the production of large crystals of apo galectin-3C suitable for neutron diffraction are described. Five data sets have been collected at three different neutron sources from galectin-3C crystals of various volumes. It was possible to merge two of these to generate an almost complete neutron data set for the galectin-3C–lactose complex. These data sets provide insights into the crystal volumes and data-collection times necessary for the same system at sources with different technologies and data-collection strategies, and these insights are applicable to other systems. International Union of Crystallography 2016-10-28 /pmc/articles/PMC5108347/ /pubmed/27841752 http://dx.doi.org/10.1107/S2059798316015540 Text en © Manzoni et al. 2016 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 Manzoni, Francesco Saraboji, Kadhirvel Sprenger, Janina Kumar, Rohit Noresson, Ann-Louise Nilsson, Ulf J. Leffler, Hakon Fisher, S. Zoë Schrader, Tobias E. Ostermann, Andreas Coates, Leighton Blakeley, Matthew P. Oksanen, Esko Logan, Derek T. Perdeuteration, crystallization, data collection and comparison of five neutron diffraction data sets of complexes of human galectin-3C |
title | Perdeuteration, crystallization, data collection and comparison of five neutron diffraction data sets of complexes of human galectin-3C |
title_full | Perdeuteration, crystallization, data collection and comparison of five neutron diffraction data sets of complexes of human galectin-3C |
title_fullStr | Perdeuteration, crystallization, data collection and comparison of five neutron diffraction data sets of complexes of human galectin-3C |
title_full_unstemmed | Perdeuteration, crystallization, data collection and comparison of five neutron diffraction data sets of complexes of human galectin-3C |
title_short | Perdeuteration, crystallization, data collection and comparison of five neutron diffraction data sets of complexes of human galectin-3C |
title_sort | perdeuteration, crystallization, data collection and comparison of five neutron diffraction data sets of complexes of human galectin-3c |
topic | Research Papers |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5108347/ https://www.ncbi.nlm.nih.gov/pubmed/27841752 http://dx.doi.org/10.1107/S2059798316015540 |
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