Perdeuteration, crystallization, data collection and comparison of five neutron diffraction data sets of complexes of human galectin-3C
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- 作者：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 and Derek T. Logan
- 关键词：neutron crystallography ; galectin-3C ; perdeuteration ; crystallogenesis
- 刊名：Acta Crystallographica Section D
- 出版年：2016
- 出版时间：November 2016
- 年：2016
- 卷：72
- 期：11
- 页码：1194-1202
- 全文大小：1436K
- ISSN：1399-0047
文摘
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.