Facile synthesis of oligosaccharide probes for the analysis of protein-carbohydrate interactions.
A strategy for facile oligosaccharide synthesis is described. It obviates chromatographic separation of intermediates and enables the isolation of desired oligomers with capture-release purification and reverse-phase silica-gel cartridge separation. As an example, preparation of monoglucosylated mannotetraose (Glc alpha1-->3Man alpha1-->2Man1alpha-->2Man1alpha-->3Man beta) was conducted. After sequential glycosylation, capture-release purification with Cys-resin, global deprotection, and reverse-phase silica-gel cartridge separation, the target pentasaccharide was isolated, while isolation of shorter oligomers that lack nonreducing-end residues (Man alpha1-->2Man1alpha-->2Man1alpha-->3Man beta, Man1alpha-->2Man1alpha-->3Man beta, Man1alpha-->3Man beta) was also achieved. These products were connected to a thiol-containing linker and immobilized on Au-coated chips. Their affinity to recombinant calreticulin was evaluated by quartz-crystal microbalance.
Takatani
Maki
M
RIKEN (The Institute of Physical and Chemical Research), 2-1 Hirosawa, Wako, Saitama 351-0198, Japan.
Ito
Yukishige
Y
D037282:Calreticulin / Q000737:chemistry*; D002241:Carbohydrates / Q000737:chemistry*; D006031:Glycosylation; D015335:Molecular Probes
D037282:Calreticulin; D002241:Carbohydrates; D015335:Molecular Probes