Monitoring the N-glycosylation of plant glycoproteins by fluorophore-assisted carbohydrate electrophoresis

• Published in: Electrophoresis Vol. 21; no. 12; pp. 2550 - 2556
• Main Authors: Bardor, Muriel, Cabanes-Macheteau, Marion, Faye, Loïc, Lerouge, Patrice
• Format: Journal Article
• Language: English
• Published: Hoboken Wiley Subscription Services, Inc., A Wiley Company 01.07.2000
• Subjects: 6-trisulfonic acid labeled plant N-glycans; 8-Aminonaphthalene-1; 8‐Aminonaphthalene‐1,3,6‐trisulfonic acid labeled plant N‐glycans; Carbohydrate Sequence; Electrophoresis, Polyacrylamide Gel - methods; Fluorescent Dyes; Fluorophore-assisted carbohydrate electrophoresis; Glycoproteins - analysis; Glycosylation; Mannose; Molecular Sequence Data; Naphthalenes; Plant glycoproteins; Plant Proteins - analysis; Polysaccharides - analysis
• ISSN: 0173-0835; 1522-2683
• DOI: 10.1002/1522-2683(20000701)21:12<2550::AID-ELPS2550>3.0.CO;2-G

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.