Enzymatic modification of heparan sulfate on a biochip promotes its interaction with antithrombin III

Biochem Biophys Res Commun. 2000 Sep 16;276(1):292-7. doi: 10.1006/bbrc.2000.3453.


A heparan sulfate glycosaminoglycan chain, biotinylated at its reducing-end, was bound to a streptavidin-coated biochip. Surface plasmon resonance spectroscopy showed a low affinity interaction with antithrombin III (ATIII) when it was flowed over a surface containing heparan sulfate. ATIII bound tightly with high affinity when the same surface was enzymatically modified to using 3-O-sulfotransferase isoform 1 (3-OST-1) in the presence of 3'-phosphoadenosine 5'-phosphosulfate (PAPS). The 3-OST-1 enzyme is involved in heparan sulfate biosynthesis and introduces a critical 3-O-sulfo group into this glycosaminoglycan affording the appropriate pentasaccharide sequence capable of high affinity binding to ATIII. This experiment demonstrates the specific structural modification of a glycosaminoglycan bound to a biochip using a biosynthetic enzyme, suggesting a new approach to rapid screening glycosaminoglycan-protein interactions.

Publication types

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

MeSH terms

  • Animals
  • Antithrombin III / chemistry*
  • Antithrombin III / metabolism
  • Binding Sites
  • Heparitin Sulfate / chemistry*
  • Heparitin Sulfate / metabolism
  • Mice
  • Oligosaccharides
  • Protein Binding


  • Oligosaccharides
  • Antithrombin III
  • Heparitin Sulfate