Perdeuteration, crystallization, data collection and comparison of five neutron diffraction data sets of complexes of human galectin-3C
(2016) In Acta Crystallographica Section D: Structural Biology 72(11). p.1194-1202- Abstract
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... (More)
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.Perdeuteration, purification and the growth of large crystals of the carbohydrate-recognition domain of galectin-3C are described. Five neutron diffraction data sets have been collected at four neutron sources; these are compared and two are merged.
(Less)
- author
- Manzoni, Francesco
LU
; Kadhirvel, Saraboji
LU
; Sprenger, Janina
LU
; Kumar, Rohit
LU
; Noresson, Ann Louise
LU
; Nilsson, Ulf J.
LU
; Leffler, Hakon
LU
; Fisher, S. Zoë
; Schrader, Tobias E.
; Ostermann, Andreas
; Coates, Leighton
; Blakeley, Matthew P.
; Oksanen, Esko
LU
and Logan, Derek T.
LU
(Less) - organization
- publishing date
- 2016-11-01
- type
- Contribution to journal
- publication status
- published
- subject
- keywords
- crystallogenesis, galectin-3C, neutron crystallography, perdeuteration
- in
- Acta Crystallographica Section D: Structural Biology
- volume
- 72
- issue
- 11
- pages
- 9 pages
- publisher
- John Wiley & Sons Inc.
- external identifiers
-
- pmid:27841752
- wos:000387593100004
- scopus:84995896935
- ISSN
- 2059-7983
- DOI
- 10.1107/S2059798316015540
- language
- English
- LU publication?
- yes
- id
- e9812300-6158-46be-be30-4b8ba2818c5f
- date added to LUP
- 2016-12-05 12:58:56
- date last changed
- 2024-01-04 18:02:36
@article{e9812300-6158-46be-be30-4b8ba2818c5f,
abstract = {{<p>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.Perdeuteration, purification and the growth of large crystals of the carbohydrate-recognition domain of galectin-3C are described. Five neutron diffraction data sets have been collected at four neutron sources; these are compared and two are merged.</p>}},
author = {{Manzoni, Francesco and Kadhirvel, Saraboji and Sprenger, Janina and Kumar, Rohit and Noresson, Ann Louise and Nilsson, Ulf J. and Leffler, Hakon and Fisher, S. Zoë and Schrader, Tobias E. and Ostermann, Andreas and Coates, Leighton and Blakeley, Matthew P. and Oksanen, Esko and Logan, Derek T.}},
issn = {{2059-7983}},
keywords = {{crystallogenesis; galectin-3C; neutron crystallography; perdeuteration}},
language = {{eng}},
month = {{11}},
number = {{11}},
pages = {{1194--1202}},
publisher = {{John Wiley & Sons Inc.}},
series = {{Acta Crystallographica Section D: Structural Biology}},
title = {{Perdeuteration, crystallization, data collection and comparison of five neutron diffraction data sets of complexes of human galectin-3C}},
url = {{http://dx.doi.org/10.1107/S2059798316015540}},
doi = {{10.1107/S2059798316015540}},
volume = {{72}},
year = {{2016}},
}