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

Forskningsoutput: TidskriftsbidragArtikel i vetenskaplig tidskrift

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.

Detaljer

Författare
Enheter & grupper
Externa organisationer
  • Jülich Research Centre
  • Technical University of Munich
  • Oak Ridge National Laboratory
  • Institut Laue Langevin
  • Los Alamos National Laboratory
Forskningsområden

Ämnesklassifikation (UKÄ) – OBLIGATORISK

  • Strukturbiologi

Nyckelord

Originalspråkengelska
Sidor (från-till)1194-1202
Antal sidor9
TidskriftActa Crystallographica Section D: Structural Biology
Volym72
Utgivningsnummer11
StatusPublished - 2016 nov 1
PublikationskategoriForskning
Peer review utfördJa