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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. As a result, 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.

Authors:
 [1];  [2];  [3];  [3];  [3]; ORCiD logo [3];  [3];  [4];  [5];  [6]; ORCiD logo [7]; ORCiD logo [8];  [1];  [3]
  1. Lund Univ., Lund (Sweden); European Spallation Source ERIC, Lund (Sweden)
  2. Lund Univ., Lund (Sweden); SASTRA Univ., Tamil Nadu (India)
  3. Lund Univ., Lund (Sweden)
  4. Los Alamos National Lab. (LANL), Los Alamos, NM (United States); European Spallation Source ERIC, Lund (Sweden)
  5. Forschungszentrum Julich GmbH, Garching (Germany)
  6. Technische Univ. Munchen, Garching (Germany)
  7. Oak Ridge National Lab. (ORNL), Oak Ridge, TN (United States)
  8. Institut Laue-Langevin, Grenoble (France)
Publication Date:
Research Org.:
Oak Ridge National Lab. (ORNL), Oak Ridge, TN (United States)
Sponsoring Org.:
USDOE
OSTI Identifier:
1494022
Grant/Contract Number:  
AC05-00OR22725
Resource Type:
Accepted Manuscript
Journal Name:
Acta Crystallographica. Section D. Structural Biology
Additional Journal Information:
Journal Volume: 72; Journal Issue: 11; Journal ID: ISSN 2059-7983
Publisher:
IUCr
Country of Publication:
United States
Language:
English
Subject:
59 BASIC BIOLOGICAL SCIENCES; neutron crystallography; galectin-3C; perdeuteration; crystallogenesis

Citation Formats

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, and Logan, Derek T. Perdeuteration, crystallization, data collection and comparison of five neutron diffraction data sets of complexes of human galectin-3C. United States: N. p., 2016. Web. doi:10.1107/S2059798316015540.
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. United States. doi:10.1107/S2059798316015540.
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, and Logan, Derek T. Fri . "Perdeuteration, crystallization, data collection and comparison of five neutron diffraction data sets of complexes of human galectin-3C". United States. doi:10.1107/S2059798316015540. https://www.osti.gov/servlets/purl/1494022.
@article{osti_1494022,
title = {Perdeuteration, crystallization, data collection and comparison of five neutron diffraction data sets of complexes of human galectin-3C},
author = {Manzoni, Francesco and Saraboji, Kadhirvel 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.},
abstractNote = {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. As a result, 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.},
doi = {10.1107/S2059798316015540},
journal = {Acta Crystallographica. Section D. Structural Biology},
number = 11,
volume = 72,
place = {United States},
year = {2016},
month = {10}
}

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Cited by: 9 works
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Figures / Tables:

Figure 1 Figure 1: (a) SDS–PAGE gel of purified galectin-3C. (b) A typical crystal obtained by repeated macroseeding. (c) The 1.8 mm3 crystal obtained by repeated feeding of a sitting drop that was used to collect data to 1.7 Å resolution at LADI-III.

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