Crystallization and crystal manipulation of the Pterocarpus angolensis seed lectin.

TitleCrystallization and crystal manipulation of the Pterocarpus angolensis seed lectin.
Publication TypeJournal Article
Year of Publication2005
AuthorsLoris, R., A. Garcia-Pino, L. Buts, J. Bouckaert, S. Beeckmans, H. De Greve, and L. Wyns
JournalActa Crystallogr D Biol Crystallogr
Volume61
IssuePt 6
Pagination685-9
Date Published2005 Jun
ISSN0907-4449
KeywordsCrystallography, X-Ray, Disaccharidases, Mannose-Binding Lectins, Plant Proteins, Protein Structure, Tertiary, Pterocarpus, Seeds
Abstract

The Man/Glc-specific legume lectin from the seeds of the African bloodwood tree (Pterocarpus angolensis) was crystallized in the presence of the disaccharide ligand Man(alpha1-3)ManMe. Small crystals initially appeared from a preliminary screen, but proved difficult to reproduce. The initial crystals were used to prepare microseeds, leading to a reproducible crystallization protocol. All attempts to obtain crystals directly of the ligand-free protein or of other carbohydrate complexes failed. However, the Man(alpha1-3)ManMe co-crystals withstand soaking with ten other carbohydrates known to bind to the lectin. Soaking for 15 min in 100 mM carbohydrate typically resulted in complete replacement of Man(alpha1-3)ManMe by the desired carbohydrate despite the involvement of lattice contacts at the binding site. Transferring the crystals for two weeks in carbohydrate-free artificial mother liquor resulted in the complete removal of the sugar from one of the two monomers in the asymmetric unit. Additional treatment of these crystals with 100 mM EDTA for two weeks resulted in removal of the structural calcium and manganese ions, which is accompanied by significant structural rearrangements of the loops that constitute the carbohydrate-binding site.

DOI10.1107/S0907444904028707
Alternate JournalActa Crystallogr. D Biol. Crystallogr.
PubMed ID15930620