Identification of ectonucleotide pyrophosphatase/phosphodiesterase 3 (ENPP3) as a regulator of N-acetylglucosaminyltransferase GnT-IX (GnT-Vb)

J Biol Chem. 2013 Sep 27;288(39):27912-26. doi: 10.1074/jbc.M113.474304. Epub 2013 Aug 19.


Our previous studies on a β1,6-N-acetylglucosaminyltransferase, GnT-IX (GnT-Vb), a homolog of GnT-V, indicated that the enzyme has a broad GlcNAc transfer activity toward N-linked and O-mannosyl glycan core structures and that its brain-specific gene expression is regulated by epigenetic histone modifications. In this study, we demonstrate the existence of an endogenous inhibitory factor for GnT-IX that functions as a key regulator for GnT-IX enzymatic activity in Neuro2a (N2a) cells. We purified this factor from N2a cells and found that it is identical to ectonucleotide pyrophosphatase/phosphodiesterase 3 (ENPP3), as evidenced by mass spectrometry and by the knockdown and overexpression of ENPP3 in cultured cells. Kinetic analyses revealed that the mechanism responsible for the inhibition of GnT-IX caused by ENPP3 is the ENPP3-mediated hydrolysis of the nucleotide sugar donor substrate, UDP-GlcNAc, with the resulting generation of UMP, a potent and competitive inhibitor of GnT-IX. Indeed, ENPP3 knockdown cells had significantly increased levels of intracellular nucleotide sugars and displayed changes in the total cellular glycosylation profile. In addition to chaperones or other known regulators of glycosyltransferases, the ENPP3-mediated hydrolysis of nucleotide sugars would have widespread and significant impacts on glycosyltransferase activities and would be responsible for altering the total cellular glycosylation profile and modulating cellular functions.

Keywords: Glycobiology; Glycosylation; Glycosyltransferases; Nucleoside Nucleotide Metabolism; Phosphodiesterases.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • COS Cells
  • Carbohydrates / chemistry
  • Chlorocebus aethiops
  • Gene Expression Regulation, Enzymologic*
  • Glycosylation
  • Glycosyltransferases / metabolism
  • HEK293 Cells
  • Humans
  • Hydrolysis
  • Mice
  • N-Acetylglucosaminyltransferases / metabolism*
  • Nerve Tissue Proteins / metabolism*
  • Nucleosides / chemistry
  • Phosphoric Diester Hydrolases / metabolism*
  • Plasmids / metabolism
  • Pyrophosphatases / metabolism*
  • RNA Interference


  • Carbohydrates
  • ENPP3 protein, human
  • Nerve Tissue Proteins
  • Nucleosides
  • Glycosyltransferases
  • MGAT5B protein, human
  • N-Acetylglucosaminyltransferases
  • Phosphoric Diester Hydrolases
  • Pyrophosphatases