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Enzymatic Synthesis of Heparan Sulfate and Heparin

  • April Joice
  • Karthik Raman
  • Caitlin Mencio
  • Maritza V. Quintero
  • Spencer Brown
  • Thao Kim Nu Nguyen
  • Balagurunathan KuberanEmail author
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 1229)

Abstract

Heparan sulfate (HS) polysaccharide chains have been shown to orchestrate distinct biological functions in several systems. Study of HS structure-function relations is, however, hampered due to the lack of availability of HS in sufficient quantities as well as the molecular heterogeneity of naturally occurring HS. Enzymatic synthesis of HS is an attractive alternative to the use of naturally occurring HS, as it reduces molecular heterogeneity, or a long and daunting chemical synthesis of HS. Heparosan, produced by E. coli K5 bacteria, has a structure similar to the unmodified HS backbone structure and can be used as a precursor in the enzymatic synthesis of HS-like polysaccharides. Here, we describe an enzymatic approach to synthesize several specifically sulfated HS polysaccharides for biological studies using the heparosan backbone and a combination of recombinant biosynthetic enzymes such as C5-epimerase and sulfotransferases.

Key words

Glycosaminoglycan Proteoglycan Heparin Heparan sulfate Enzymatic synthesis Epimerase Sulfotransferase 

Notes

Acknowledgement

This work was supported in part by NIH grants (P01HL107152 and R01GM075168) to B.K. and by the NIH fellowship F31CA168198 to K.R.

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Copyright information

© Springer Science+Business Media New York 2015

Authors and Affiliations

  • April Joice
    • 1
  • Karthik Raman
    • 2
  • Caitlin Mencio
    • 3
  • Maritza V. Quintero
    • 1
  • Spencer Brown
    • 1
  • Thao Kim Nu Nguyen
    • 1
    • 4
  • Balagurunathan Kuberan
    • 1
    • 2
    • 3
    Email author
  1. 1.Department of Medicinal ChemistryUniversity of UtahSalt Lake CityUSA
  2. 2.Department of BioengineeringUniversity of UtahSalt Lake CityUSA
  3. 3.Interdepartmental Program in NeuroscienceUniversity of UtahSalt Lake CityUSA
  4. 4.Institute of Microbiology and BiotechnologyVietnam National UniversityHanoiVietnam

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