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Enzymatic synthesis of antithrombin III–binding heparan sulfate pentasaccharide

Nature Biotechnology volume 21, pages 1343–1346 (2003)Cite this article

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.

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Figure 1: ATIII-binding heparan sulfate pentasaccharide.
Figure 2: Synthesis of ATIII-binding heparan sulfate pentasaccharide.
Figure 3: Structural and functional analysis of synthetic pentasaccharide 7.

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Acknowledgements

We thank Irvin Segel for his generous gift of APS kinase. We thank Keiichi Yoshida, Seikagaku Corporation, for generously providing us with Δ4,5-glycuronidase.

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Authors and Affiliations

  1. Department of Biology, Massachusetts Institute of Technology, Cambridge, 02139, Massachusetts, USA

    Balagurunathan Kuberan, Miroslaw Z Lech, David L Beeler, Zhengliang L Wu & Robert D Rosenberg

  2. Division of Molecular and Vascular Medicine, Beth Israel Deaconess Medical Center, Harvard Medical School, Boston, 02215, Massachusetts, USA

    Robert D Rosenberg

Authors
  1. Balagurunathan Kuberan
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  2. Miroslaw Z Lech
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  3. David L Beeler
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  4. Zhengliang L Wu
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  5. Robert D Rosenberg
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Correspondence to Robert D Rosenberg.

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R.D.R. has recently, with the agreement and participation of the Massachusetts Institute of Technology, established a company (Carbogenix) to commercialize the technology described in this paper.

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Kuberan, B., Lech, M., Beeler, D. et al. Enzymatic synthesis of antithrombin III–binding heparan sulfate pentasaccharide. Nat Biotechnol 21, 1343–1346 (2003). https://doi.org/10.1038/nbt885

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