Identification of N-acetylglucosamine-alpha-1-phosphate transferase activity in Dictyostelium discoideum: an enzyme that initiates phosphoglycosylation

Biochem Biophys Res Commun. 1995 Mar 8;208(1):384-9. doi: 10.1006/bbrc.1995.1349.

Abstract

A lysosomal proteinase from Dictyostelium discoideum was previously shown to have GlcNAc alpha-1-P residues in phosphodiester linkage to serine. We have identified a GlcNAc-alpha-1-P transferase activity in membrane preparations using UDP[3H]GlcNAc and a peptide acceptor with three tandem Ser-Gly repeats. We established an assay, proved the structure of the product, determined the Kms for donor and acceptor and showed that the glycopeptide binds a GlcNAc-alpha-1-P specific rabbit antibody. These findings provide the tools to search for mutants lacking GlcNAc-alpha-1-P transferase activity as a probe for the function of this modification we call phosphoglycosylation.

Publication types

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

MeSH terms

  • Acetylglucosamine / analogs & derivatives
  • Acetylglucosamine / analysis
  • Animals
  • Chromatography, Affinity
  • Chromatography, High Pressure Liquid
  • Chromatography, Ion Exchange
  • Chromatography, Thin Layer
  • Dictyostelium / enzymology*
  • Glycosylation
  • Immunoglobulin G
  • Lysosomes / enzymology
  • Rabbits / immunology
  • Transferases (Other Substituted Phosphate Groups) / metabolism*

Substances

  • Immunoglobulin G
  • N-acetylglucosamine-1-phosphate
  • Transferases (Other Substituted Phosphate Groups)
  • UDP-N-acetylglucosamine-lysosomal-enzyme-N-acetylglucosaminephosphotransferase
  • Acetylglucosamine