2-Iminothiolane: a reagent for the introduction of sulphydryl groups into oligosaccharides derived from asparagine-linked glycans.
A facile method for introducing reactive sulphydryl groups into oligosaccharides was developed. 1-Amino-oligosaccharides 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.
Tarentino
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Division of Clinical Sciences, Wadsworth Center for Laboratories and Research, New York State Department of Health, Albany 12201-0509.
Phelan
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AW
Plummer
T H
TH
D001216:Asparagine; D002236:Carbohydrate Conformation; D002240:Carbohydrate Sequence; D002851:Chromatography, High Pressure Liquid; D002852:Chromatography, Ion Exchange; D003432:Cross-Linking Reagents; D005340:Fibrinogen / Q000737:chemistry*; D005456:Fluorescent Dyes; D006020:Glycopeptides / Q000737:chemistry; D006801:Humans; D007096:Imidoesters; D007075:Immunoglobulin M / Q000737:chemistry*; D007202:Indicators and Reagents; D007700:Kinetics; D008969:Molecular Sequence Data; D009282:Naphthalenesulfonates; D009844:Oligosaccharides / Q000737:chemistry* / Q000302:isolation & purification; D011134:Polysaccharides; D013438:Sulfhydryl Compounds; D000509:alpha-Fetoproteins / Q000737:chemistry
D003432:Cross-Linking Reagents; D005456:Fluorescent Dyes; D006020:Glycopeptides; D007096:Imidoesters; D007075:Immunoglobulin M; D007202:Indicators and Reagents; D009282:Naphthalenesulfonates; D009844:Oligosaccharides; D011134:Polysaccharides; D013438:Sulfhydryl Compounds; D000509:alpha-Fetoproteins; C005905:methyl 4-mercaptobutyrimidate; C006421:1,5-I-AEDANS; D001216:Asparagine; D005340:Fibrinogen