Sulfation of N-acetylglucosamine by Chondroitin 6-sulfotransferase 2 (GST-5)

J Biol Chem. 2000 Dec 22;275(51):40226-34. doi: 10.1074/jbc.M006414200.

Abstract

Based on sequence homology with a previously cloned human GlcNAc 6-O-sulfotransferase, we have identified an open reading frame (ORF) encoding a novel member of the Gal/GalNAc/GlcNAc 6-O-sulfotransferase (GST) family termed GST-5 on the human X chromosome (band Xp11). GST-5 has recently been characterized as a novel GalNAc 6-O-sulfotransferase termed chondroitin 6-sulfotransferase-2 (Kitagawa, H., Fujita, M., Itio, N., and Sugahara K. (2000) J. Biol. Chem. 275, 21075-21080). We have coexpressed a human GST-5 cDNA with a GlyCAM-1/IgG fusion protein in COS-7 cells and observed four-fold enhanced [(35)S]sulfate incorporation into this mucin acceptor. All mucin-associated [(35)S]sulfate was incorporated as GlcNAc-6-sulfate or Galbeta1-->4GlcNAc-6-sulfate. GST-5 was also expressed in soluble epitope-tagged form and found to catalyze 6-O-sulfation of GlcNAc residues in synthetic acceptor structures. In particular, GST-5 was found to catalyze 6-O-sulfation of beta-benzyl GlcNAc but not alpha- or beta-benzyl GalNAc. In the mouse genome we have found a homologous ORF that predicts a novel murine GlcNAc 6-O-sulfotransferase with 88% identity to the human enzyme. This gene was mapped to mouse chromosome X at band XA3.1-3.2. GST-5 is the newest member of an emerging family of carbohydrate 6-O-sulfotransferases that includes chondroitin 6-sulfotransferase (GST-0), keratan-sulfate galactose 6-O-sulfotransferase (GST-1), the ubiquitously expressed GlcNAc 6-O-sulfotransferase (GST-2), high endothelial cell GlcNAc 6-O-sulfotransferase (GST-3), and intestinal GlcNAc 6-O-sulfotransferase (GST-4).

Publication types

  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Acetylglucosamine / chemistry*
  • Amino Acid Sequence
  • Animals
  • Base Sequence
  • Catalysis
  • Chromosome Mapping
  • DNA Primers
  • DNA, Complementary
  • Isoenzymes / genetics
  • Isoenzymes / metabolism*
  • Mice
  • Molecular Sequence Data
  • Open Reading Frames
  • Sulfates / chemistry*
  • Sulfotransferases / genetics
  • Sulfotransferases / metabolism*
  • X Chromosome

Substances

  • DNA Primers
  • DNA, Complementary
  • Isoenzymes
  • Sulfates
  • Sulfotransferases
  • carbohydrate sulfotransferases
  • Acetylglucosamine

Associated data

  • GENBANK/AF280089