Heparan sulfate synthesized by mouse embryonic stem cells deficient in NDST1 and NDST2 is 6-O-sulfated but contains no N-sulfate groups

Research output: Contribution to journalArticle

Abstract

Heparan sulfate structure differs significantly between various cell types and during different developmental stages. The diversity is created during biosynthesis by sulfotransferases, which add sulfate groups to the growing chain, and a C5-epimerase, which converts selected glucuronic acid residues to iduronic acid. All these modifications are believed to depend on initial glucosamine N-sulfation carried out by the enzyme glucosaminyl N-deacetylase/N-sulfotransferase (NDST). Here we report that heparan sulfate synthesized by mouse embryonic stem cells deficient in NDST1 and NDST2 completely lacks N-sulfation but still contains 6-O-sulfate groups, demonstrating that 6-O-sulfation can occur without prior N-sulfation. Reverse transcriptase-PCR analysis indicates that all three identified 6-O-sulfotransferases are expressed by the cells, 6-O-sulfotransferase-1 being the dominating form. The 6-O-sulfated polysaccharide lacking N-sulfate groups also contains N-unsubstituted glucosamine units, raising questions about how these units are generated.

Details

Authors
  • Katarina Holmborn
  • Johan Ledin
  • Emanuel Smeds
  • Inger Eriksson
  • Marion Kusche-Gullberg
  • Lena Kjellén
External organisations
  • Uppsala University
Research areas and keywords

Subject classification (UKÄ) – MANDATORY

  • Cell and Molecular Biology

Keywords

  • Amidohydrolases/genetics, Animals, Blastocyst/metabolism, Carbohydrate Epimerases/chemistry, Chromatography, High Pressure Liquid, Chromatography, Ion Exchange, DNA, Complementary/metabolism, Embryo, Mammalian/metabolism, Gene Expression Regulation, Developmental, Genotype, Glucosamine/chemistry, Glucuronic Acid/metabolism, Glycosaminoglycans, Heparitin Sulfate/biosynthesis, Iduronic Acid/metabolism, Mice, Mice, Transgenic, Nitrous Acid/metabolism, Polysaccharides/chemistry, Reverse Transcriptase Polymerase Chain Reaction, Stem Cells/metabolism, Sulfates/chemistry, Sulfotransferases/genetics, Sulfur/metabolism
Original languageEnglish
Pages (from-to)42355-8
JournalJournal of Biological Chemistry
Volume279
Issue number41
Publication statusPublished - 2004
Publication categoryResearch
Peer-reviewedYes
Externally publishedYes