Expression of heparan sulphate L-iduronyl 2-O-sulphotransferase in human kidney 293 cells results in increased D-glucuronyl 2-O-sulphation

Biochem J. 2000 Mar 1;346 Pt 2(Pt 2):463-8.

Abstract

Functionally important interactions between heparan sulphate and a variety of proteins depend on the precise location of O-sulphate groups. Such residues occur at C-2 of L-iduronic (IdoA) and D-glucuronic acid (GlcA) units, and at C-3 and C-6 of D-glucosamine (GlcN) units. Stable transfection of human embryonic kidney 293 cells with a cDNA encoding mouse mastocytoma IdoA 2-O-sulphotransferase resulted in an approx. 6-fold increase in O-sulphotransferase activity, compared with control cells, as determined using O-desulphated heparin as an acceptor. Structural analysis of endogenous heparan sulphate in the transfected cells, following metabolic labelling with either [(3)H]GlcN or [(35)S]sulphate, showed appreciable formation of -GlcA(2-OSO(3))-GlcNSO(3)- disaccharide units (6% of total disaccharide units; 17% of total O-sulphated disaccharide units) that were essentially absent from heparan sulphate from control cells. The increase in GlcA 2-O-sulphation was accompanied by a decrease in the amount of IdoA formed, whereas overall 2-O-sulphation or 6-O-sulphation remained largely unaffected. These findings indicate that 2-O-sulphation of IdoA and GlcA residues is catalysed by the same enzyme in heparan sulphate biosynthesis.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Carbohydrate Epimerases / metabolism*
  • Cell Line
  • Glucuronates / metabolism
  • Heparan Sulfate Proteoglycans / metabolism*
  • Humans
  • Kidney / metabolism*
  • Mice
  • Molecular Sequence Data
  • Sulfotransferases / metabolism*

Substances

  • Glucuronates
  • Heparan Sulfate Proteoglycans
  • Sulfotransferases
  • heparan sulfate L-iduronyl 2-O-sulfotransferase
  • Carbohydrate Epimerases

Associated data

  • GENBANK/AF169243