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M211480200v1
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Papers In Press, published online ahead of print January 2, 2003
J. Biol. Chem, 10.1074/jbc.M211480200
Submitted on November 11, 2002
Revised on January 2, 2003
Accepted on January 2, 2003

beta 1,4-galactosyltransferase(beta 4GalT)-IV is specific for GlcNAc 6-O-sulfate : beta 4GalT-IV acts on keratan sulfate-related glycans and a precursor glycan of 6-sulfo sialyl Lewis X

Akira Seko, Naoshi Dohmae, Koji Takio, and Katsuko Yamashita

Biochemistry, Sasaki Institute, Tokyo 101-0062

Corresponding Author: yamashita{at}sasaki.or.jp

The galbeta 1-4(SO3--6)GlcNAc moiety is present in various N-linked and O-linked glycans including keratan sulfate and 6-sulfo-sialyl Lewis X, a L-selectin ligand. We previously found lbeta 1,4galactosyltransferase(lbeta 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 lbeta 4GalT has a similar sequence to human lbeta 4GalT-IV. To confirm this result, we prepared cDNA for each of the seven beta 4GalTs cloned to date and examined substrate specificities using the membrane fractions derived from beta 4GalT-transfected COS-7 cells. When using several N-linked and O-linked glycans with or without 6SGN residues as acceptor substrates, only beta 4GalT-IV efficiently recognized 6SGN, keratan sulfate-related oligosaccharides, and Gallbeta 1-3((SO3--6GlcNAcbeta 1-6)GalNAcalpha 1-O-pNP, a precursor for 6-sulfo-sialyl Lewis X. These results suggested that beta 4GalT-IV is a 6SGN-specific beta 4GalT and may be involved in the biosynthesis of various glycoproteins carrying a 6-O-sulfated N-acetyllactosamine moiety.


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