Cloning of a human UDP-N-acetyl-α-D-galactosamine:Polypeptide N- acetylgalactosaminyltransferase that complements other GalNAc-transferases in complete O-glycosylation of the MUC1 tandem repeat

Eric Paul Bennett, Helle Hassan, Ulla Mandel, Ekatarina Mirgorodskaya, Peter Roepstorff, Joy Burchell, Joyce Taylor-Papadimitriou, Michael A Hollingsworth, Gerard Merkx, Ad Geurts Van Kessel, Hans Eiberg, Rudi Steffensen, Henrik Clausen

Research output: Contribution to journalArticle

186 Citations (Scopus)

Abstract

A fourth human UDP-GalNAc:polypeptide N-acetyl- galactosaminyltransferase, designated GalNAc-T4, was cloned and expressed. The genomic organization of GalNAc-T4 is distinct from GalNAc-T1, -T2, and - T3, which contain multiple coding exons, in that the coding region is contained in a single exon. GalNAc-T4 was placed at human chromosome 12q21.3- q22 by in situ hybridization and linkage analysis. GalNAc-T4 expressed in Sf9 cells or in a stably transfected Chinese hamster ovary cell line exhibited a unique acceptor substrate specificity. GalNAc-T4 transferred GalNAc to two sites in the MUC1 tandem repeat sequence (Ser in GVTSA and Thr in PDTR) using a 24-mer glycopeptide with GalNAc residues attached at sites utilized by GalNAc-T1, -T2, and -T3 (TAPPAHGVTSAPDTRPAPGSTAPPA, GalNAc attachment sites underlined). Furthermore, GalNAc-T4 showed the best kinetic properties with an O-glycosylation site in the P-selectin glycoprotein ligand-1 molecule. Northern analysis of human organs revealed a wide expression pattern. Immunohistology with a monoclonal antibody showed the expected Golgi-like localization in salivary glands. A single base polymorphism, G1516A (Val to Ile), was identified (allele frequency 34%). The function of GalNAc-T4 complements other GalNAc-transferases in O-glycosylation of MUC1 showing that glycosylation of MUC1 is a highly ordered process and changes in the repertoire or topology of GalNAc-transferases will result in altered pattern of O-glycan attachments.

Original languageEnglish (US)
Pages (from-to)30472-30481
Number of pages10
JournalJournal of Biological Chemistry
Volume273
Issue number46
DOIs
StatePublished - Nov 13 1998

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Glycosylation
Acetylgalactosamine
Tandem Repeat Sequences
Uridine Diphosphate
Cloning
Organism Cloning
polypeptide N-acetylgalactosaminyltransferase
Exons
Glycopeptides
Sf9 Cells
Chromosomes
Polymorphism
Human Chromosomes
Polysaccharides
Substrate Specificity
Cricetulus
Salivary Glands
Gene Frequency

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology
  • Cell Biology

Cite this

Cloning of a human UDP-N-acetyl-α-D-galactosamine:Polypeptide N- acetylgalactosaminyltransferase that complements other GalNAc-transferases in complete O-glycosylation of the MUC1 tandem repeat. / Bennett, Eric Paul; Hassan, Helle; Mandel, Ulla; Mirgorodskaya, Ekatarina; Roepstorff, Peter; Burchell, Joy; Taylor-Papadimitriou, Joyce; Hollingsworth, Michael A; Merkx, Gerard; Van Kessel, Ad Geurts; Eiberg, Hans; Steffensen, Rudi; Clausen, Henrik.

In: Journal of Biological Chemistry, Vol. 273, No. 46, 13.11.1998, p. 30472-30481.

Research output: Contribution to journalArticle

Bennett, EP, Hassan, H, Mandel, U, Mirgorodskaya, E, Roepstorff, P, Burchell, J, Taylor-Papadimitriou, J, Hollingsworth, MA, Merkx, G, Van Kessel, AG, Eiberg, H, Steffensen, R & Clausen, H 1998, 'Cloning of a human UDP-N-acetyl-α-D-galactosamine:Polypeptide N- acetylgalactosaminyltransferase that complements other GalNAc-transferases in complete O-glycosylation of the MUC1 tandem repeat', Journal of Biological Chemistry, vol. 273, no. 46, pp. 30472-30481. https://doi.org/10.1074/jbc.273.46.30472
Bennett, Eric Paul ; Hassan, Helle ; Mandel, Ulla ; Mirgorodskaya, Ekatarina ; Roepstorff, Peter ; Burchell, Joy ; Taylor-Papadimitriou, Joyce ; Hollingsworth, Michael A ; Merkx, Gerard ; Van Kessel, Ad Geurts ; Eiberg, Hans ; Steffensen, Rudi ; Clausen, Henrik. / Cloning of a human UDP-N-acetyl-α-D-galactosamine:Polypeptide N- acetylgalactosaminyltransferase that complements other GalNAc-transferases in complete O-glycosylation of the MUC1 tandem repeat. In: Journal of Biological Chemistry. 1998 ; Vol. 273, No. 46. pp. 30472-30481.
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title = "Cloning of a human UDP-N-acetyl-α-D-galactosamine:Polypeptide N- acetylgalactosaminyltransferase that complements other GalNAc-transferases in complete O-glycosylation of the MUC1 tandem repeat",
abstract = "A fourth human UDP-GalNAc:polypeptide N-acetyl- galactosaminyltransferase, designated GalNAc-T4, was cloned and expressed. The genomic organization of GalNAc-T4 is distinct from GalNAc-T1, -T2, and - T3, which contain multiple coding exons, in that the coding region is contained in a single exon. GalNAc-T4 was placed at human chromosome 12q21.3- q22 by in situ hybridization and linkage analysis. GalNAc-T4 expressed in Sf9 cells or in a stably transfected Chinese hamster ovary cell line exhibited a unique acceptor substrate specificity. GalNAc-T4 transferred GalNAc to two sites in the MUC1 tandem repeat sequence (Ser in GVTSA and Thr in PDTR) using a 24-mer glycopeptide with GalNAc residues attached at sites utilized by GalNAc-T1, -T2, and -T3 (TAPPAHGVTSAPDTRPAPGSTAPPA, GalNAc attachment sites underlined). Furthermore, GalNAc-T4 showed the best kinetic properties with an O-glycosylation site in the P-selectin glycoprotein ligand-1 molecule. Northern analysis of human organs revealed a wide expression pattern. Immunohistology with a monoclonal antibody showed the expected Golgi-like localization in salivary glands. A single base polymorphism, G1516A (Val to Ile), was identified (allele frequency 34{\%}). The function of GalNAc-T4 complements other GalNAc-transferases in O-glycosylation of MUC1 showing that glycosylation of MUC1 is a highly ordered process and changes in the repertoire or topology of GalNAc-transferases will result in altered pattern of O-glycan attachments.",
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T1 - Cloning of a human UDP-N-acetyl-α-D-galactosamine:Polypeptide N- acetylgalactosaminyltransferase that complements other GalNAc-transferases in complete O-glycosylation of the MUC1 tandem repeat

AU - Bennett, Eric Paul

AU - Hassan, Helle

AU - Mandel, Ulla

AU - Mirgorodskaya, Ekatarina

AU - Roepstorff, Peter

AU - Burchell, Joy

AU - Taylor-Papadimitriou, Joyce

AU - Hollingsworth, Michael A

AU - Merkx, Gerard

AU - Van Kessel, Ad Geurts

AU - Eiberg, Hans

AU - Steffensen, Rudi

AU - Clausen, Henrik

PY - 1998/11/13

Y1 - 1998/11/13

N2 - A fourth human UDP-GalNAc:polypeptide N-acetyl- galactosaminyltransferase, designated GalNAc-T4, was cloned and expressed. The genomic organization of GalNAc-T4 is distinct from GalNAc-T1, -T2, and - T3, which contain multiple coding exons, in that the coding region is contained in a single exon. GalNAc-T4 was placed at human chromosome 12q21.3- q22 by in situ hybridization and linkage analysis. GalNAc-T4 expressed in Sf9 cells or in a stably transfected Chinese hamster ovary cell line exhibited a unique acceptor substrate specificity. GalNAc-T4 transferred GalNAc to two sites in the MUC1 tandem repeat sequence (Ser in GVTSA and Thr in PDTR) using a 24-mer glycopeptide with GalNAc residues attached at sites utilized by GalNAc-T1, -T2, and -T3 (TAPPAHGVTSAPDTRPAPGSTAPPA, GalNAc attachment sites underlined). Furthermore, GalNAc-T4 showed the best kinetic properties with an O-glycosylation site in the P-selectin glycoprotein ligand-1 molecule. Northern analysis of human organs revealed a wide expression pattern. Immunohistology with a monoclonal antibody showed the expected Golgi-like localization in salivary glands. A single base polymorphism, G1516A (Val to Ile), was identified (allele frequency 34%). The function of GalNAc-T4 complements other GalNAc-transferases in O-glycosylation of MUC1 showing that glycosylation of MUC1 is a highly ordered process and changes in the repertoire or topology of GalNAc-transferases will result in altered pattern of O-glycan attachments.

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