Characterization of heparan sulfate N-deacetylase/N-sulfotransferase isoform 4 using synthetic oligosaccharide substrates

Biochim Biophys Acta Gen Subj. 2018 Mar;1862(3):547-556. doi: 10.1016/j.bbagen.2017.11.016. Epub 2017 Nov 21.


Background: The final structure of heparan sulfate chains is strictly regulated in vivo, though the biosynthesis is not guided by a template process. N-deacetylase/N-sulfotransferase (NDST) is the first modification enzyme in the HS biosynthetic pathway. The N-sulfo groups introduced by NDST are reportedly involved in determination of the susceptibility to subsequent processes catalyzed by C5-epimerse and 3-O-sulfotransferases. Understanding the substrate specificities of the four human NDST isoforms has become central to uncovering the regulatory mechanism of HS biosynthesis.

Methods: Highly-purified recombinant NDST-4 (rNDST-4) and a selective library of structurally-defined oligosaccharides were employed to determine the substrate specificity of rNDST-4.

Results: Full-length rNDST-4 lacks obvious N-deacetylase activity, and displays only N-sulfotransferase activity. Unlike NDST-1, NDST-4 did not show directional N-sulfotransferase activity while the N-deacetylase domain was inactive.

Conclusion and general significance: Individual NDST-4 could not effectively assume the key role in the distribution of N-S domains and N-Ac domains in HS biosynthesis in vivo.

Keywords: Biosynthesis; Catalytic mode; Heparan sulfate; N-deacetylase/N-sulfotransferase; Regulatory mechanism.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Carbohydrate Conformation
  • Carbohydrate Sequence
  • Catalysis
  • Glycosylation
  • Humans
  • Membrane Proteins / metabolism*
  • Nucleopolyhedroviruses
  • Oligosaccharides / chemical synthesis
  • Oligosaccharides / metabolism*
  • Protein Domains
  • Protein Isoforms
  • Protein Processing, Post-Translational
  • Recombinant Proteins / chemistry
  • Recombinant Proteins / metabolism
  • Small Molecule Libraries
  • Spodoptera
  • Substrate Specificity
  • Sulfotransferases / metabolism*
  • Surface Plasmon Resonance
  • Tandem Mass Spectrometry


  • Membrane Proteins
  • Oligosaccharides
  • Protein Isoforms
  • Recombinant Proteins
  • Small Molecule Libraries
  • Sulfotransferases
  • NDST4 protein, human