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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...

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Autores principales: 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.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: International Union of Crystallography 2016
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.
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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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