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

Electrophoresis. 2000 Jul;21(12):2550-6. doi: 10.1002/1522-2683(20000701)21:12<2550::AID-ELPS2550>3.0.CO;2-G.

Abstract

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.

MeSH terms

  • Carbohydrate Sequence
  • Electrophoresis, Polyacrylamide Gel / methods
  • Fluorescent Dyes*
  • Glycoproteins / analysis*
  • Glycosylation
  • Mannose
  • Molecular Sequence Data
  • Naphthalenes*
  • Plant Proteins / analysis*
  • Polysaccharides / analysis*

Substances

  • Fluorescent Dyes
  • Glycoproteins
  • Naphthalenes
  • Plant Proteins
  • Polysaccharides
  • 8-amino-1,3,6-naphthalenetrisulfonic acid
  • Mannose