2-Iminothiolane - A Reagent For The Introduction Of Sulfhydryl-Groups Into Oligosaccharides Derived From Asparagine-Linked Glycans

A facile method for introducing reactive sulphydryl groups into oligosaccharides was developed. 1-Amino-oligosacchandes generated from asparagine-linked glycans by peptide-N4(N-acetyl-beta-D-glucosaminyl) asparagine amidase (PNGase F) digestion were monitored by high-performance anion-exchange chromatography with pulsed amperometric detection and derivatized under optimal conditions with 2-iminothiolane-HCl. The resulting mercapto-butyramido oligosaccharides, which were obtained in high yield, were alkylated with a fluorescent reagent and used to selectively assay for endoglycosidases that hydrolyse di-N-acetylchitobiose linkages.