Electrophoretic resolution and fluorescence detection of N-linked glycoprotein oligosaccharides after reductive amination with 8-aminonaphthalene-1,3,6-trisulphonic acid.
The relative mobilities of various N-linked oligosaccharides reductively aminated to the charged fluorophore 8-amino-naphthalene-1,3,6-trisulphonic acid (ANTS) were determined by electrophoresis on high-density polyacrylamide slab gels. Each ANTS-derivatized oligosaccharide was assigned a relative migration index (RMI) expressed in terms of glucose equivalents, which was conveniently estimated by reference to a homologous series of ANTS--maltooligosaccharides run on each gel as oligosaccharide size standards. High-mannose-, complex- and hybrid-type structures were generally well resolved and easily visualized at picomole levels by simple UV light excitation. Application of these methods for the qualitative analysis of the oligosaccharides released from bovine fetuin and bovine asialofetuin by peptide-N-glycosidase F illustrates the usefulness of these techniques as fast, simple, and inexpensive tools for the characterization of N-linked oligosaccharides attached to glycoproteins.
Glycomed, Incorporated, Alameda, CA 94501.
R J
RJ
Stack
M T
MT
Sullivan
D000581:Amidohydrolases / Q000378:metabolism; D000818:Animals; D001212:Asialoglycoproteins / Q000032:analysis / Q000378:metabolism; D002236:Carbohydrate Conformation; D002417:Cattle; D004591:Electrophoresis, Polyacrylamide Gel; D060748:Fetuins; D006023:Glycoproteins / Q000737:chemistry*; D006868:Hydrolysis; D007700:Kinetics; D009281:Naphthalenes; D009844:Oligosaccharides / Q000032:analysis / Q000737:chemistry* / Q000378:metabolism; D043524:Peptide-N4-(N-acetyl-beta-glucosaminyl) Asparagine Amidase; D000509:alpha-Fetoproteins / Q000032:analysis / Q000378:metabolism
D001212:Asialoglycoproteins; D060748:Fetuins; D006023:Glycoproteins; D009281:Naphthalenes; D009844:Oligosaccharides; D000509:alpha-Fetoproteins; C017029:asialofetuin; C046465:8-amino-1,3,6-naphthalenetrisulfonic acid; D000581:Amidohydrolases; D043524:Peptide-N4-(N-acetyl-beta-glucosaminyl) Asparagine Amidase