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A more recent version of this article appeared on December 15, 2000
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M006414200v1
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Papers In Press, published online ahead of print August 23, 2000
J. Biol. Chem, 10.1074/jbc.M006414200
Submitted on July 19, 2000
Accepted on August 22, 2000

Sulfation of N-Acetylglucosamine by Chondroitin 6-Sulfotransferase 2 (GST-5)

Sunil Bhakta, Alexander Bartes, Kendra G. Bowman, Wei-Ming Kao, Irene Polsky, Jin K. Lee, Brian N. Cook, Richard Bruehl, Steven D. Rosen, Carolyn R. Bertozzi, and Stefan Hemmerich

Respiratory Diseases, Roche Bioscience, Palo Alto, CA 94304

Corresponding Author: stefan.hemmerich{at}roche.com

Based on sequence homology with a previously cloned human high endothelial cell N-acetylglucosamine 6-O-sulfotransferase, we have identified an open reading frame encoding a novel member of the galactose/N-acetylgalactosamine/N-acetylglucosamine sulfotransferase (GST) family termed GST-5 on the human X chromosome (band Xp11). A cDNA encoding the same protein has recently been shown to encode a novel N-acetylgalactosamine 6-O-sulfotransferase termed chondroitin 6-sulfotransferase-2 (Kitagawa et al., J. Biol. Chem. 275:21075-21080). We have coexpressed a human GST-5 cDNA with recombinant GlyCAM-1/IgG fusion protein in COS-7 cells and observed four-fold enhanced [35S]-sulfate incorporation into this mucin acceptor. Analysis of the mucin-associated carbohydrate revealed that all [35S]-sulfate was incorporated as GlcNAc-6-sulfate or Galbeta 1-4GlcNAc-6-sulfate. The putative catalytic domain of 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 beta -benzyl GalNAc or alpha -benzyl GlcNAc. In the mouse genome we have found a homologous open reading frame that predicts a novel murine N-acetylglucosamine-6-sulfotransferase that on the amino acid level is 88 % identical to the human enzyme at the amino acid level. The murine GST-5 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 N-acetylglucosamine 6-sulfotransferase (GST-3), and intestinal N-acetylglucosamine 6-sulfotransferase (GST-4).


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