Control of O-glycan branch formation: Molecular cloning of human cDNA encoding a novel β1,6-N-acetylglucosaminyltransferase forming core 2 and core 4

Tilo Schwientek, Mitsuharu Nomoto, Steven B. Levery, Gerard Merkx, Ad Geurts Van Kessel, Eric P. Bennett, Michael A Hollingsworth, Henrik Clausen

Research output: Contribution to journalArticle

92 Citations (Scopus)

Abstract

A novel human UDP-GlcNAc:Gal/GlcNAcβ1-3GalNAcα β1,6GlcNAc- transferase, designated C2/4GnT, was identified by BLAST analysis of expressed sequence tags. The sequence of C2/4GnT encoded a putative type II transmembrane protein with significant sequence similarity to human C2GnT and IGnT. Expression of the secreted form of C2/4GnT in insect cells showed that the gene product had UDP-N-acetyl-α-D-glucosamine:acceptor β1,6-N- acetylglucosaminyltransferase (β1,6GlcNAc-transferase) activity. Analysis of substrate specificity revealed that the enzyme catalyzed O-glycan branch formation of the core 2 and core 4 type. NMR analyses of the product formed with core 3-para-nitrophenyl confirmed the product core 4-para-nitrophenyl. The coding region of C2/4GnT was contained in a single exon and located to chromosome 15q21.3. Northern analysis revealed a restricted expression pattern of C2/4GnT mainly in colon, kidney, pancreas, and small intestine. No expression of C2/4GnT was detected in brain, heart, liver, ovary, placenta, spleen, thymus, and peripheral blood leukocytes. The expression of core 2 O- glycans has been correlated with cell differentiation processes and cancer. The results confirm the predicted existence of a β1,6GlcNAc-transferase that functions in both core 2 and core 4 O-glycan branch formation. The redundancy in β1,6GlcNAc-transferases capable of forming core 2 O-glycans is important for understanding the mechanisms leading to specific changes in core 2 branching during cell development and malignant transformation.

Original languageEnglish (US)
Pages (from-to)4504-4512
Number of pages9
JournalJournal of Biological Chemistry
Volume274
Issue number8
DOIs
StatePublished - Feb 19 1999

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Cloning
Molecular Cloning
Transferases
Polysaccharides
Complementary DNA
Uridine Diphosphate
Thymus
Acetylglucosamine
Expressed Sequence Tags
Chromosomes
Substrate Specificity
Liver
Thymus Gland
Placenta
Small Intestine
Redundancy
Insects
Cell Differentiation
Pancreas
Ovary

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology
  • Cell Biology

Cite this

Control of O-glycan branch formation : Molecular cloning of human cDNA encoding a novel β1,6-N-acetylglucosaminyltransferase forming core 2 and core 4. / Schwientek, Tilo; Nomoto, Mitsuharu; Levery, Steven B.; Merkx, Gerard; Van Kessel, Ad Geurts; Bennett, Eric P.; Hollingsworth, Michael A; Clausen, Henrik.

In: Journal of Biological Chemistry, Vol. 274, No. 8, 19.02.1999, p. 4504-4512.

Research output: Contribution to journalArticle

Schwientek, Tilo ; Nomoto, Mitsuharu ; Levery, Steven B. ; Merkx, Gerard ; Van Kessel, Ad Geurts ; Bennett, Eric P. ; Hollingsworth, Michael A ; Clausen, Henrik. / Control of O-glycan branch formation : Molecular cloning of human cDNA encoding a novel β1,6-N-acetylglucosaminyltransferase forming core 2 and core 4. In: Journal of Biological Chemistry. 1999 ; Vol. 274, No. 8. pp. 4504-4512.
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abstract = "A novel human UDP-GlcNAc:Gal/GlcNAcβ1-3GalNAcα β1,6GlcNAc- transferase, designated C2/4GnT, was identified by BLAST analysis of expressed sequence tags. The sequence of C2/4GnT encoded a putative type II transmembrane protein with significant sequence similarity to human C2GnT and IGnT. Expression of the secreted form of C2/4GnT in insect cells showed that the gene product had UDP-N-acetyl-α-D-glucosamine:acceptor β1,6-N- acetylglucosaminyltransferase (β1,6GlcNAc-transferase) activity. Analysis of substrate specificity revealed that the enzyme catalyzed O-glycan branch formation of the core 2 and core 4 type. NMR analyses of the product formed with core 3-para-nitrophenyl confirmed the product core 4-para-nitrophenyl. The coding region of C2/4GnT was contained in a single exon and located to chromosome 15q21.3. Northern analysis revealed a restricted expression pattern of C2/4GnT mainly in colon, kidney, pancreas, and small intestine. No expression of C2/4GnT was detected in brain, heart, liver, ovary, placenta, spleen, thymus, and peripheral blood leukocytes. The expression of core 2 O- glycans has been correlated with cell differentiation processes and cancer. The results confirm the predicted existence of a β1,6GlcNAc-transferase that functions in both core 2 and core 4 O-glycan branch formation. The redundancy in β1,6GlcNAc-transferases capable of forming core 2 O-glycans is important for understanding the mechanisms leading to specific changes in core 2 branching during cell development and malignant transformation.",
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AU - Levery, Steven B.

AU - Merkx, Gerard

AU - Van Kessel, Ad Geurts

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AB - A novel human UDP-GlcNAc:Gal/GlcNAcβ1-3GalNAcα β1,6GlcNAc- transferase, designated C2/4GnT, was identified by BLAST analysis of expressed sequence tags. The sequence of C2/4GnT encoded a putative type II transmembrane protein with significant sequence similarity to human C2GnT and IGnT. Expression of the secreted form of C2/4GnT in insect cells showed that the gene product had UDP-N-acetyl-α-D-glucosamine:acceptor β1,6-N- acetylglucosaminyltransferase (β1,6GlcNAc-transferase) activity. Analysis of substrate specificity revealed that the enzyme catalyzed O-glycan branch formation of the core 2 and core 4 type. NMR analyses of the product formed with core 3-para-nitrophenyl confirmed the product core 4-para-nitrophenyl. The coding region of C2/4GnT was contained in a single exon and located to chromosome 15q21.3. Northern analysis revealed a restricted expression pattern of C2/4GnT mainly in colon, kidney, pancreas, and small intestine. No expression of C2/4GnT was detected in brain, heart, liver, ovary, placenta, spleen, thymus, and peripheral blood leukocytes. The expression of core 2 O- glycans has been correlated with cell differentiation processes and cancer. The results confirm the predicted existence of a β1,6GlcNAc-transferase that functions in both core 2 and core 4 O-glycan branch formation. The redundancy in β1,6GlcNAc-transferases capable of forming core 2 O-glycans is important for understanding the mechanisms leading to specific changes in core 2 branching during cell development and malignant transformation.

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