Monitoring the N-glycosylation of plant glycoproteins by fluorophore-assisted carbohydrate electrophoresis.
We have evaluated the efficiency of a fast, simple and efficient method, fluorophore-assisted carbohydrate electrophoresis (FACE), for the characterization of plant N-linked glycans. After their enzymatic release from plant glycoproteins, N-glycans were reductively aminated to the charged fluorophore 8-aminonaphthalene-1, 3, 6-trisulfonic acid (ANTS) and separated using high resolution polyacrylamide gel electrophoresis. In addition, an affinity purification procedure using concanavalin A was developed for separation of ANTS-labeled high-mannose-type N-glycans from other plant oligosaccharides.
Laboratoire des Transports Intracellulaires, CNRS ESA, Université de Rouen, Faculté des Sciences, Mont Saint Aignan, France.
M
M
Bardor
M
M
Cabanes-Macheteau
L
L
Faye
P
P
Lerouge
D002240:Carbohydrate Sequence; D004591:Electrophoresis, Polyacrylamide Gel / Q000379:methods; D005456:Fluorescent Dyes; D006023:Glycoproteins / Q000032:analysis*; D006031:Glycosylation; D008358:Mannose; D008969:Molecular Sequence Data; D009281:Naphthalenes; D010940:Plant Proteins / Q000032:analysis*; D011134:Polysaccharides / Q000032:analysis*
D005456:Fluorescent Dyes; D006023:Glycoproteins; D009281:Naphthalenes; D010940:Plant Proteins; D011134:Polysaccharides; C046465:8-amino-1,3,6-naphthalenetrisulfonic acid; D008358:Mannose