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.
authors.affiliation: Biochemistry and Structural Biology, Department of Chemistry, Lund University, S-221 00 Lund, Sweden.
authors.forename: Francesco
authors.initials: F
authors.lastname: Manzoni
authors.affiliation: Biochemistry and Structural Biology, Department of Chemistry, Lund University, S-221 00 Lund, Sweden.
authors.forename: Kadhirvel
authors.initials: K
authors.lastname: Saraboji
authors.affiliation: Biochemistry and Structural Biology, Department of Chemistry, Lund University, S-221 00 Lund, Sweden.
authors.forename: Janina
authors.initials: J
authors.lastname: Sprenger
authors.affiliation: Biochemistry and Structural Biology, Department of Chemistry, Lund University, S-221 00 Lund, Sweden.
authors.forename: Rohit
authors.initials: R
authors.lastname: Kumar
authors.affiliation: Centre for Analysis and Synthesis, Department of Chemistry, Lund University, S-221 00 Lund, Sweden.
authors.forename: Ann Louise
authors.initials: AL
authors.lastname: Noresson
authors.affiliation: Centre for Analysis and Synthesis, Department of Chemistry, Lund University, S-221 00 Lund, Sweden.
authors.forename: Ulf J
authors.initials: UJ
authors.lastname: Nilsson
authors.affiliation: Department of Laboratory Medicine, Section MIG, Lund University, S-221 00 Lund, Sweden.
authors.forename: Hakon
authors.initials: H
authors.lastname: Leffler
authors.affiliation: Los Alamos National Laboratory, Los Alamos, New Mexico, USA.
authors.forename: S Zoë
authors.initials: SZ
authors.lastname: Fisher
authors.affiliation: Jülich Centre for Neutron Science (JCNS) at Heinz Maier-Leibnitz Zentrum (MLZ), Forschungszentrum Jülich GmbH, Lichtenbergstrasse 1, 85748 Garching, Germany.
authors.forename: Tobias E
authors.initials: TE
authors.lastname: Schrader
authors.affiliation: Heinz Maier-Leibnitz Zentrum (MLZ), Technische Universität München, Lichtenbergstrasse 1, 85748 Garching, Germany.
authors.forename: Andreas
authors.initials: A
authors.lastname: Ostermann
authors.affiliation: Biology and Soft Matter Division, Oak Ridge National Laboratory, Oak Ridge, Tennessee, USA.
authors.forename: Leighton
authors.initials: L
authors.lastname: Coates
authors.affiliation: Institut Laue-Langevin, 71 Avenue des Martyrs, 38000 Grenoble, France.
authors.forename: Matthew P
authors.initials: MP
authors.lastname: Blakeley
authors.affiliation: Biochemistry and Structural Biology, Department of Chemistry, Lund University, S-221 00 Lund, Sweden.
authors.forename: Esko
authors.initials: E
authors.lastname: Oksanen
authors.affiliation: Biochemistry and Structural Biology, Department of Chemistry, Lund University, S-221 00 Lund, Sweden.
authors.forename: Derek T
authors.initials: DT
authors.lastname: Logan
mesh_terms: D001798:Blood Proteins; D003460:Crystallization / Q000379:methods; D003903:Deuterium / Q000737:chemistry; D037502:Galectin 3 / Q000737:chemistry* / Q000378:metabolism; D037161:Galectins; D006801:Humans; D007785:Lactose / Q000737:chemistry / Q000378:metabolism; D008958:Models, Molecular; D033363:Neutron Diffraction / Q000379:methods*; D011487:Protein Conformation
chemical_list: D001798:Blood Proteins; D037502:Galectin 3; D037161:Galectins; C428445:LGALS3 protein, human; D003903:Deuterium; D007785:Lactose
keywords: crystallogenesis; galectin-3C; neutron crystallography; perdeuteration