Molecular Cloning and Expression of a Novel Human β -Gal-3-O-sulfotransferase that Acts Preferentially on N-Acetyllactosamine in N- and O-Glycans

Atsushi Suzuki, Nobuyoshi Hiraoka, Masami Suzuki, Kiyohiko Angata, Anup K. Misra, Joseph McAuliffe, Ole Hindsgaul, Minoru Fukuda

Research output: Contribution to journalArticle

27 Citations (Scopus)

Abstract

A novel cDNA-encoding galactose 3-O-sulfotransferase was cloned by screening the expressed sequence tag data base using the previously cloned cDNA encoding a galactosyl ceramide 3-O-sulfotransferase, which we term Gal3ST-1. The newly isolated cDNA encodes a novel 3-O-sulfotransferase, termed Gal3ST-3, that acts exclusively on N-acetyllactosamine present in N-glycans and core2-branched O-glycans. These conclusions were confirmed by analyzing CD43 chimeric proteins in Chinese hamster ovary cells expressing core2 β1,6-N-acetylglucosaminyltransferase. The acceptor specificity of Gal3ST-3 contrasts with that of the recently cloned galactose 3-O-sulfotransferase (Honke, K., Tsuda, M., Koyota, S., Wada, Y., Iida-Tanaka, N., Ishizuka, I., Nakayama, J., and Taniguchi, N. (2001) J. Biol. Chem. 276, 267-274), which we term Gal3ST-2 in the present study because the latter enzyme can also act on core1 O-glycan and type 1 oligosaccharides, Galβ1→3GlcNAc. Moreover, Gal3ST-3 but not Gal3ST-2 can act on Galβ1→4(sulfo→6)GlcNAc, indicating that disulfated sulfo→3Galβ1→4(sulfo→6) GlcNAc→R may be formed by Gal3ST-3 in combination with GlcNAc 6-O-sulfotransferase. Although both Gal3ST-2 and Gal3ST-3 do not act on galactosyl ceramide, Gal3ST-3 is only moderately more homologous to Gal3ST-2 (40.1%) than to Gal3ST-1 (38.0%) at the amino acid level. Northern blot analysis demonstrated that transcripts for Gal3ST-3 are predominantly expressed in the brain, kidney, and thyroid where the presence of 3′-sulfation of N-acetyllactosamine has been reported. These results indicate that the newly cloned Gal3ST-3 plays a critical role in 3′-sulfation of N-acetyllactosamine in both O- and N-glycans.

Original languageEnglish
Pages (from-to)24388-24395
Number of pages8
JournalJournal of Biological Chemistry
Volume276
Issue number26
DOIs
Publication statusPublished - 2001 Jun 29
Externally publishedYes

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Sulfotransferases
Cloning
Molecular Cloning
Polysaccharides
Galactosylceramides
Complementary DNA
Expressed Sequence Tags
Cricetulus
Oligosaccharides
Northern Blotting
Ovary
Brain
Screening
Thyroid Gland
Cells
Databases
Kidney
Amino Acids
N-acetyllactosamine
Enzymes

ASJC Scopus subject areas

  • Biochemistry

Cite this

Molecular Cloning and Expression of a Novel Human β -Gal-3-O-sulfotransferase that Acts Preferentially on N-Acetyllactosamine in N- and O-Glycans. / Suzuki, Atsushi; Hiraoka, Nobuyoshi; Suzuki, Masami; Angata, Kiyohiko; Misra, Anup K.; McAuliffe, Joseph; Hindsgaul, Ole; Fukuda, Minoru.

In: Journal of Biological Chemistry, Vol. 276, No. 26, 29.06.2001, p. 24388-24395.

Research output: Contribution to journalArticle

Suzuki, Atsushi ; Hiraoka, Nobuyoshi ; Suzuki, Masami ; Angata, Kiyohiko ; Misra, Anup K. ; McAuliffe, Joseph ; Hindsgaul, Ole ; Fukuda, Minoru. / Molecular Cloning and Expression of a Novel Human β -Gal-3-O-sulfotransferase that Acts Preferentially on N-Acetyllactosamine in N- and O-Glycans. In: Journal of Biological Chemistry. 2001 ; Vol. 276, No. 26. pp. 24388-24395.
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abstract = "A novel cDNA-encoding galactose 3-O-sulfotransferase was cloned by screening the expressed sequence tag data base using the previously cloned cDNA encoding a galactosyl ceramide 3-O-sulfotransferase, which we term Gal3ST-1. The newly isolated cDNA encodes a novel 3-O-sulfotransferase, termed Gal3ST-3, that acts exclusively on N-acetyllactosamine present in N-glycans and core2-branched O-glycans. These conclusions were confirmed by analyzing CD43 chimeric proteins in Chinese hamster ovary cells expressing core2 β1,6-N-acetylglucosaminyltransferase. The acceptor specificity of Gal3ST-3 contrasts with that of the recently cloned galactose 3-O-sulfotransferase (Honke, K., Tsuda, M., Koyota, S., Wada, Y., Iida-Tanaka, N., Ishizuka, I., Nakayama, J., and Taniguchi, N. (2001) J. Biol. Chem. 276, 267-274), which we term Gal3ST-2 in the present study because the latter enzyme can also act on core1 O-glycan and type 1 oligosaccharides, Galβ1→3GlcNAc. Moreover, Gal3ST-3 but not Gal3ST-2 can act on Galβ1→4(sulfo→6)GlcNAc, indicating that disulfated sulfo→3Galβ1→4(sulfo→6) GlcNAc→R may be formed by Gal3ST-3 in combination with GlcNAc 6-O-sulfotransferase. Although both Gal3ST-2 and Gal3ST-3 do not act on galactosyl ceramide, Gal3ST-3 is only moderately more homologous to Gal3ST-2 (40.1{\%}) than to Gal3ST-1 (38.0{\%}) at the amino acid level. Northern blot analysis demonstrated that transcripts for Gal3ST-3 are predominantly expressed in the brain, kidney, and thyroid where the presence of 3′-sulfation of N-acetyllactosamine has been reported. These results indicate that the newly cloned Gal3ST-3 plays a critical role in 3′-sulfation of N-acetyllactosamine in both O- and N-glycans.",
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AU - Angata, Kiyohiko

AU - Misra, Anup K.

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N2 - A novel cDNA-encoding galactose 3-O-sulfotransferase was cloned by screening the expressed sequence tag data base using the previously cloned cDNA encoding a galactosyl ceramide 3-O-sulfotransferase, which we term Gal3ST-1. The newly isolated cDNA encodes a novel 3-O-sulfotransferase, termed Gal3ST-3, that acts exclusively on N-acetyllactosamine present in N-glycans and core2-branched O-glycans. These conclusions were confirmed by analyzing CD43 chimeric proteins in Chinese hamster ovary cells expressing core2 β1,6-N-acetylglucosaminyltransferase. The acceptor specificity of Gal3ST-3 contrasts with that of the recently cloned galactose 3-O-sulfotransferase (Honke, K., Tsuda, M., Koyota, S., Wada, Y., Iida-Tanaka, N., Ishizuka, I., Nakayama, J., and Taniguchi, N. (2001) J. Biol. Chem. 276, 267-274), which we term Gal3ST-2 in the present study because the latter enzyme can also act on core1 O-glycan and type 1 oligosaccharides, Galβ1→3GlcNAc. Moreover, Gal3ST-3 but not Gal3ST-2 can act on Galβ1→4(sulfo→6)GlcNAc, indicating that disulfated sulfo→3Galβ1→4(sulfo→6) GlcNAc→R may be formed by Gal3ST-3 in combination with GlcNAc 6-O-sulfotransferase. Although both Gal3ST-2 and Gal3ST-3 do not act on galactosyl ceramide, Gal3ST-3 is only moderately more homologous to Gal3ST-2 (40.1%) than to Gal3ST-1 (38.0%) at the amino acid level. Northern blot analysis demonstrated that transcripts for Gal3ST-3 are predominantly expressed in the brain, kidney, and thyroid where the presence of 3′-sulfation of N-acetyllactosamine has been reported. These results indicate that the newly cloned Gal3ST-3 plays a critical role in 3′-sulfation of N-acetyllactosamine in both O- and N-glycans.

AB - A novel cDNA-encoding galactose 3-O-sulfotransferase was cloned by screening the expressed sequence tag data base using the previously cloned cDNA encoding a galactosyl ceramide 3-O-sulfotransferase, which we term Gal3ST-1. The newly isolated cDNA encodes a novel 3-O-sulfotransferase, termed Gal3ST-3, that acts exclusively on N-acetyllactosamine present in N-glycans and core2-branched O-glycans. These conclusions were confirmed by analyzing CD43 chimeric proteins in Chinese hamster ovary cells expressing core2 β1,6-N-acetylglucosaminyltransferase. The acceptor specificity of Gal3ST-3 contrasts with that of the recently cloned galactose 3-O-sulfotransferase (Honke, K., Tsuda, M., Koyota, S., Wada, Y., Iida-Tanaka, N., Ishizuka, I., Nakayama, J., and Taniguchi, N. (2001) J. Biol. Chem. 276, 267-274), which we term Gal3ST-2 in the present study because the latter enzyme can also act on core1 O-glycan and type 1 oligosaccharides, Galβ1→3GlcNAc. Moreover, Gal3ST-3 but not Gal3ST-2 can act on Galβ1→4(sulfo→6)GlcNAc, indicating that disulfated sulfo→3Galβ1→4(sulfo→6) GlcNAc→R may be formed by Gal3ST-3 in combination with GlcNAc 6-O-sulfotransferase. Although both Gal3ST-2 and Gal3ST-3 do not act on galactosyl ceramide, Gal3ST-3 is only moderately more homologous to Gal3ST-2 (40.1%) than to Gal3ST-1 (38.0%) at the amino acid level. Northern blot analysis demonstrated that transcripts for Gal3ST-3 are predominantly expressed in the brain, kidney, and thyroid where the presence of 3′-sulfation of N-acetyllactosamine has been reported. These results indicate that the newly cloned Gal3ST-3 plays a critical role in 3′-sulfation of N-acetyllactosamine in both O- and N-glycans.

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