Endoglycosidase and glycoamidase release of N-linked glycans

Curr Protoc Immunol. 2010 Apr:Chapter 8:8.15.1-8.15.25. doi: 10.1002/0471142735.im0815s89.

Abstract

Nearly all proteins entering the lumen of the endoplasmic reticulum (ER) become glycosylated en route to a cellular organelle, the plasma membrane, or the extracellular space. Many glycans can be attached to proteins, but the most common are the N-linked glycans (oligosaccharides). These chains are added very soon after a protein enters the ER, but they undergo extensive remodeling (processing), especially in the Golgi. Processing changes the sensitivity of the N-glycan to enzymes that cleave entire sugar chains or individual monosaccharides, which also changes the migration of the protein on SDS gels. These changes can be used to indicate when a protein has passed a particular subcellular location. This unit details some of the methods used to track a protein as it trafficks from the ER to the Golgi toward its final location.

MeSH terms

  • Acetylglucosamine / chemistry
  • Amidohydrolases / chemistry*
  • Carbohydrate Conformation
  • Electrophoresis, Polyacrylamide Gel
  • Electrophoretic Mobility Shift Assay
  • Endoplasmic Reticulum / metabolism*
  • Glycoproteins / chemistry*
  • Glycoproteins / metabolism
  • Glycosaminoglycans / chemistry
  • Glycoside Hydrolases / chemistry*
  • Glycosylation
  • Golgi Apparatus / metabolism*
  • Immunoprecipitation
  • Mannose / chemistry
  • Oligosaccharides / chemistry*
  • Oligosaccharides / metabolism
  • Peptide-N4-(N-acetyl-beta-glucosaminyl) Asparagine Amidase / chemistry
  • Protein Transport

Substances

  • Glycoproteins
  • Glycosaminoglycans
  • Oligosaccharides
  • Glycoside Hydrolases
  • Amidohydrolases
  • Peptide-N4-(N-acetyl-beta-glucosaminyl) Asparagine Amidase
  • Mannose
  • Acetylglucosamine