Perdeuteration, crystallization, data collection and comparison of five neutron diffraction data sets of complexes of human galectin-3C

Francesco Manzoni; Kadhirvel Saraboji; Janina Sprenger; Rohit Kumar; Ann Louise Noresson; Ulf J. Nilsson; Hakon Leffler; S. Zoë Fisher; Tobias E. Schrader; Andreas Ostermann; Leighton Coates; Matthew P. Blakeley; Esko Oksanen; Derek T. Logan

doi:10.1107/S2059798316015540

Perdeuteration, crystallization, data collection and comparison of five neutron diffraction data sets of complexes of human galectin-3C

Francesco Manzoni, Kadhirvel Saraboji, Janina Sprenger, Rohit Kumar, Ann Louise Noresson, Ulf J. Nilsson, Hakon Leffler, S. Zoë Fisher, Tobias E. Schrader, Andreas Ostermann, Leighton Coates, Matthew P. Blakeley, Esko Oksanen, Derek T. Logan

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15 Scopus citations

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 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.

Original language	English
Pages (from-to)	1194-1202
Number of pages	9
Journal	Acta Crystallographica Section D: Structural Biology
Volume	72
Issue number	11
DOIs	https://doi.org/10.1107/S2059798316015540
State	Published - Nov 1 2016

Funding

RK was supported by a project grant from the Knut and Alice Wallenberg Foundation KAW2013.022). The work was also funded by two seed grants from the Natural Science Faculty at Lund University to DTL. Research at ORNLs Spallation Neutron Source was sponsored by the Scientific User Facilities Division, Office of Basic Energy Sciences, US Department of Energy. The Office of Biological and Environmental Research supported research at Oak Ridge National Laboratorys Center for Structural Molecular Biology (CSMB) and the Protein Crystallography Station at Los Alamos National Laboratory, using facilities supported by the Scientific User Facilities Division, Office of Basic Energy Sciences, US Department of Energy. For experiments at FRM-II, DTL and FM received funding from the European Unions Seventh Framework Programme for research, technological development and demonstration under NMI3-II Grant No. 283883.

Keywords

crystallogenesis
galectin-3C
neutron crystallography
perdeuteration

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10.1107/S2059798316015540

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Manzoni, F., Saraboji, K., Sprenger, J., Kumar, R., Noresson, A. L., Nilsson, U. J., Leffler, H., Fisher, S. Z., Schrader, T. E., Ostermann, A., Coates, L., Blakeley, M. P., Oksanen, E., & Logan, D. T. (2016). Perdeuteration, crystallization, data collection and comparison of five neutron diffraction data sets of complexes of human galectin-3C. Acta Crystallographica Section D: Structural Biology, 72(11), 1194-1202. https://doi.org/10.1107/S2059798316015540

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title = "Perdeuteration, crystallization, data collection and comparison of five neutron diffraction data sets of complexes of human galectin-3C",

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 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.",

keywords = "crystallogenesis, galectin-3C, neutron crystallography, perdeuteration",

author = "Francesco Manzoni and Kadhirvel Saraboji and Janina Sprenger and Rohit Kumar and Noresson, {Ann Louise} and Nilsson, {Ulf J.} and Hakon Leffler and Fisher, {S. Zo{\"e}} and Schrader, {Tobias E.} and Andreas Ostermann and Leighton Coates and Blakeley, {Matthew P.} and Esko Oksanen and Logan, {Derek T.}",

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Manzoni, F, Saraboji, K, Sprenger, J, Kumar, R, Noresson, AL, Nilsson, UJ, Leffler, H, Fisher, SZ, Schrader, TE, Ostermann, A, Coates, L, Blakeley, MP, Oksanen, E & Logan, DT 2016, 'Perdeuteration, crystallization, data collection and comparison of five neutron diffraction data sets of complexes of human galectin-3C', Acta Crystallographica Section D: Structural Biology, vol. 72, no. 11, pp. 1194-1202. https://doi.org/10.1107/S2059798316015540

Perdeuteration, crystallization, data collection and comparison of five neutron diffraction data sets of complexes of human galectin-3C. / Manzoni, Francesco; Saraboji, Kadhirvel; Sprenger, Janina et al.
In: Acta Crystallographica Section D: Structural Biology, Vol. 72, No. 11, 01.11.2016, p. 1194-1202.

Research output: Contribution to journal › Article › peer-review

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T1 - Perdeuteration, crystallization, data collection and comparison of five neutron diffraction data sets of complexes of human galectin-3C

AU - Manzoni, Francesco

AU - Saraboji, Kadhirvel

AU - Sprenger, Janina

AU - Kumar, Rohit

AU - Noresson, Ann Louise

AU - Nilsson, Ulf J.

AU - Leffler, Hakon

AU - Fisher, S. Zoë

AU - Schrader, Tobias E.

AU - Ostermann, Andreas

AU - Coates, Leighton

AU - Blakeley, Matthew P.

AU - Oksanen, Esko

AU - Logan, Derek T.

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AB - 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.

KW - crystallogenesis

KW - galectin-3C

KW - neutron crystallography

KW - perdeuteration

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Perdeuteration, crystallization, data collection and comparison of five neutron diffraction data sets of complexes of human galectin-3C

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