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Synthesis of Sulfur Isotope-Labeled Sulfate Donor, 3′-Phosphoadenosine-5′-Phosphosulfate, for Studying Glycosaminoglycan Functions

  • Caitlin Mencio
  • Vimal P. Swarup
  • Marcus Soliai
  • Balagurunathan KuberanEmail author
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 1229)

Abstract

The biological activity of glycosaminoglycans (GAGs) depends greatly on the sulfation pattern present within the GAG chain. Chemical biology of GAGs can be further advanced by preparation of sulfur-isotope-enriched sulfated GAGs. 3′-Phosphoadenosine-5′-phosphosulfate (PAPS) serves as a universal sulfate donor in the sulfation of GAGs by sulfotransferases. Therefore, synthesis of PAPS carrying sulfur isotopes is critical in the preparation of labeled GAGs for biochemical studies. Here we describe a robust in vitro enzymatic synthesis of sulfur isotope-enriched PAPS which allows for heavy- or radio-isotope labeling of GAG chains.

Key words

PAPS APS kinase Sulfurylase PAPS synthase Radioactive sulfate Glycosaminoglycans 

Notes

Acknowledgement

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

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Copyright information

© Springer Science+Business Media New York 2015

Authors and Affiliations

  • Caitlin Mencio
    • 1
  • Vimal P. Swarup
    • 2
    • 3
  • Marcus Soliai
    • 2
  • Balagurunathan Kuberan
    • 1
    • 2
    • 3
    Email author
  1. 1.Interdepartmental Program in NeuroscienceUniversity of UtahSalt Lake CityUSA
  2. 2.Department of Medicinal ChemistryUniversity of UtahSalt Lake CityUSA
  3. 3.Department of BioengineeringUniversity of UtahSalt Lake CityUSA

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