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J. Biol. Chem., Vol. 278, Issue 11, 9150-9158, March 14, 2003

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1,4-Galactosyltransferase (4GalT)-IV Is Specific for GlcNAc 6-O-Sulfate
4GalT-IV ACTS ON KERATAN SULFATE-RELATED GLYCANS AND A PRECURSOR GLYCAN OF 6-SULFOSIALYL-LEWIS X^*

Akira Seko, Naoshi Dohmae^§, Koji Takio^§¶, and Katsuko Yamashita

From the  Department of Biochemistry, Sasaki Institute, Kanda-Surugadai 2-2, and Core Research for Evolutional Science and Technology (CREST) of the Japan Science and Technology Corporation, Kanda-Surugadai 2-3, Chiyoda-ku, Tokyo 101-0062, and ^§ Biomolecular Characterization Division, the Institute of Physical and Chemical Research (RIKEN), Hirosawa 2-1, Wako, Saitama 351-0198, Japan

The Gal14(SO6)GlcNAc moiety is present in various N-linked and O-linked glycans including keratan sulfate and 6-sulfosialyl-Lewis X, an L-selectin ligand. We previously found 1,4-galactosyltransferase (4GalT) activity in human colonic mucosa, which prefers GlcNAc 6-O-sulfate (6SGN) as an acceptor to non-substituted GlcNAc (Seko, A., Hara-Kuge, S., Nagata, K., Yonezawa, S., and Yamashita, K. (1998) FEBS Lett. 440, 307-310). To identify the gene for this enzyme, we purified the enzyme from porcine colonic mucosa. The purified enzyme had the characteristic requirement of basic lipids for catalytic activity. Analysis of the partial amino acid sequence of the enzyme revealed that the purified 4GalT has a similar sequence to human 4GalT-IV. To confirm this result, we prepared cDNA for each of the seven 4GalTs cloned to date and examined substrate specificities using the membrane fractions derived from 4GalT-transfected COS-7 cells. When using several N-linked and O-linked glycans with or without 6SGN residues as acceptor substrates, only 4GalT-IV efficiently recognized 6SGN, keratan sulfate-related oligosaccharides, and Gal13(SO6GlcNAc16) GalNAc1-O-pNP, a precursor for 6-sulfosialyl-Lewis X. These results suggested that 4GalT-IV is a 6SGN-specific 4GalT and may be involved in the biosynthesis of various glycoproteins carrying a 6-O-sulfated N-acetyllactosamine moiety.

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