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-sulfosialyl-Lewis X.
The Galbeta1-->4(SO(3)(-)-->6)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 beta1,4-galactosyltransferase (beta4GalT) 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 beta4GalT has a similar sequence to human beta4GalT-IV. To confirm this result, we prepared cDNA for each of the seven beta4GalTs cloned to date and examined substrate specificities using the membrane fractions derived from beta4GalT-transfected COS-7 cells. When using several N-linked and O-linked glycans with or without 6SGN residues as acceptor substrates, only beta4GalT-IV efficiently recognized 6SGN, keratan sulfate-related oligosaccharides, and Galbeta1-->3(SO(3)(-)-->6GlcNAcbeta1-->6) GalNAcalpha1-O-pNP, a precursor for 6-sulfosialyl-Lewis X. These results suggested that beta4GalT-IV is a 6SGN-specific beta4GalT and may be involved in the biosynthesis of various glycoproteins carrying a 6-O-sulfated N-acetyllactosamine moiety.
Seko
Akira
A
Department of Biochemistry, Sasaki Institute, Kanda-Surugadai 2-2, Tokyo 101-0062, Japan.
Dohmae
Naoshi
N
Takio
Koji
K
Yamashita
Katsuko
K
D000117:Acetylglucosamine / Q000031:analogs & derivatives* / Q000378:metabolism*; D000588:Amines / Q000737:chemistry; D000595:Amino Acid Sequence; D000596:Amino Acids / Q000737:chemistry; D000606:Amino Sugars / Q000737:chemistry; D000818:Animals; D015152:Blotting, Northern; D019556:COS Cells; D002240:Carbohydrate Sequence; D002845:Chromatography; D003001:Cloning, Molecular; D018076:DNA, Complementary / Q000378:metabolism; D004305:Dose-Response Relationship, Drug; D004591:Electrophoresis, Polyacrylamide Gel; D005700:Galactosyltransferases / Q000737:chemistry* / Q000378:metabolism*; D015723:Gene Library; D006801:Humans; D007632:Keratan Sulfate / Q000378:metabolism; D016256:Lewis X Antigen / Q000737:chemistry / Q000378:metabolism; D008055:Lipids; D008198:Lymph Nodes / Q000378:metabolism; D008956:Models, Chemical; D008969:Molecular Sequence Data; D010455:Peptides / Q000737:chemistry; D011134:Polysaccharides / Q000737:chemistry; D011485:Protein Binding; D017434:Protein Structure, Tertiary; D012685:Sepharose / Q000494:pharmacology; D013110:Sphingosine / Q000031:analogs & derivatives* / Q000737:chemistry; D013552:Swine; D014018:Tissue Distribution; D014162:Transfection; D014530:Uridine Diphosphate / Q000378:metabolism
D000588:Amines; D000596:Amino Acids; D000606:Amino Sugars; D018076:DNA, Complementary; D016256:Lewis X Antigen; D008055:Lipids; D010455:Peptides; D011134:Polysaccharides; C033137:N-acetylglucosamine-6-sulfate; C000458:N-acetyllactosamine; D014530:Uridine Diphosphate; D012685:Sepharose; D007632:Keratan Sulfate; D005700:Galactosyltransferases; C111678:beta-1,4-galactosyltransferase IV; C009317:stearylamine; C061800:N,N-dimethylsphingosine; D013110:Sphingosine; D000117:Acetylglucosamine