Glycosylation of nuclear pore protein p62. Reticulocyte lysate catalyzes O-linked N-acetylglucosamine addition in vitro

J Biol Chem. 1990 Apr 25;265(12):6868-73.


The addition of O-linked N-acetylglucosamine (GlcNAc) to the major nuclear pore complex glycoprotein p62 was examined. Expression of the rat p62 cDNA in transfected monkey cells was detected using a rat p62-specific antipeptide antiserum and two previously described nuclear pore-specific monoclonal antibodies which require O-linked GlcNAc for binding. Although the p62 cDNA was predicted to encode a 54-kDa polypeptide, the product expressed in monkey cells migrated on sodium dodecyl sulfate-polyacrylamide gel electrophoresis as two species of 62 and 59-kDa. Cell-free translation of the p62 in vitro transcript yielded a 59-kDa polypeptide using wheat germ extract and a 62-kDa product using a commercially available rabbit reticulocyte lysate. Several lines of evidence indicated that the 62-kDa rabbit reticulocyte lysate translation product was modified by O-linked N-acetylglucosamine; the protein bound specifically to a wheat germ agglutinin affinity column and was converted to 59 kDa when treated with jack bean beta-acetylglucosaminidase. The 59-kDa unglycosylated wheat germ translation product was converted to the 62-kDa glycosylated form upon incubation with reticulocyte lysate demonstrating that O-linked GlcNAc can be added to p62 post-translationally.

Publication types

  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Acetylglucosamine / metabolism*
  • Amino Acid Sequence
  • Animals
  • Antibodies, Monoclonal
  • Cell Line
  • Chromatography, Affinity
  • DNA / genetics
  • Fluorescent Antibody Technique
  • Glucosamine / analogs & derivatives*
  • Glycosylation
  • Membrane Glycoproteins*
  • Membrane Proteins / genetics*
  • Membrane Proteins / isolation & purification
  • Membrane Proteins / metabolism
  • Molecular Sequence Data
  • Molecular Weight
  • Nuclear Pore Complex Proteins
  • Protein Biosynthesis
  • Protein Processing, Post-Translational*
  • Rats
  • Reticulocytes / metabolism*
  • Transfection


  • Antibodies, Monoclonal
  • Membrane Glycoproteins
  • Membrane Proteins
  • Nuclear Pore Complex Proteins
  • nuclear pore protein p62
  • DNA
  • Glucosamine
  • Acetylglucosamine