Enzymatic synthesis of antithrombin III-binding heparan sulfate pentasaccharide

Nat Biotechnol. 2003 Nov;21(11):1343-6. doi: 10.1038/nbt885. Epub 2003 Oct 5.

Abstract

Heparan sulfate (HS) proteoglycans are crucial to numerous biological processes and pathological conditions, but to date only a few HS structures have been synthesized and characterized with regard to structure-function relationships. Because HS proteoglycans are highly diverse in structure, there are substantial limitations on their synthesis by classical chemical means, and thus new methods to rapidly assemble bioactive HS structures are needed. Here we report the biosynthesis of bioactive HS oligosaccharides using an engineered set of cloned enzymes that mimics the Golgi apparatus in vitro. We rapidly and efficiently assembled the antithrombin III-binding pentasaccharide in just 6 steps, in contrast to the approximately 60 steps needed for its chemical synthesis, with an overall yield at least twofold greater and a completion time at least 100 times faster than for the chemical process.

MeSH terms

  • Antithrombin III / chemistry*
  • Antithrombin III / metabolism
  • Biomimetics / methods*
  • Cloning, Molecular / methods
  • Golgi Apparatus / enzymology
  • Heparan Sulfate Proteoglycans / chemical synthesis
  • Heparan Sulfate Proteoglycans / chemistry*
  • Heparan Sulfate Proteoglycans / metabolism
  • Molecular Weight
  • Polysaccharides / chemical synthesis
  • Polysaccharides / chemistry
  • Polysaccharides / metabolism
  • Protein Binding
  • Protein Engineering / methods*
  • Racemases and Epimerases / chemistry*
  • Racemases and Epimerases / genetics
  • Racemases and Epimerases / metabolism*
  • Recombinant Proteins / chemistry
  • Recombinant Proteins / metabolism
  • Sulfotransferases / chemistry*
  • Sulfotransferases / genetics
  • Sulfotransferases / metabolism*

Substances

  • Heparan Sulfate Proteoglycans
  • Polysaccharides
  • Recombinant Proteins
  • Antithrombin III
  • Sulfotransferases
  • heparitin sulfotransferase
  • Racemases and Epimerases