Expression of GDP-L-Fuc: Gal(β1-4)GlcNAc-R (Fuc to GlcNAc) α-1,3-fucosyltransferase and its relationship to glycoprotein structure in a human erythroleukemia cell line, HEL

R. L. Giuntoli, L. I. Stoykova, D. R B Gillies, M. C. Glick

Research output: Contribution to journalArticle

Abstract

Terminal glycosylation may be a mechanism to control the function of specific biologically active glycoproteins. The biosynthesis of terminal sialyl and fucosyl residues on certain glycoproteins has been linked to the expression of the respective glycosyltransferase. In contrast, a human erythroleukemia cell line, HEL, contained a highly active GDP-L-Fuc: Gal(β1-4)GlcNAc-R (Fuc to GlcNAc) α-1,3-fucosyltransferase (α-1,3-fucosyltransferase) but no detectable α-1,3-linked fucosyI residues on the glycoproteins. The α-1,3-fucosyltransferase gave apparent K(m) values for Fuc(α1-2)Gal(β1-4)GlcNAc β-O-benzyl, Gal(β1-4)GlcNAc and GDP-fucose of 0.04, 0.68 and 0.12 mM, respectively. The lack of detectable fucosyl residues in α-1,3-linkage to GlcNAc on the [3H]fucose-labeled glycoproteins was shown with the use of almond α-1,3/4-fucosidase and internal controls to verify that the enzyme was active. Using Western-blot analysis, HEL cell glycoproteins reacted with blood group H type-2 antibody, confirming the presence of Fuc(α1-2)Gal(β1-4)GlcNAc as reported by others and the presence of the preferred substrate for the enzyme. It is proposed that controls for terminal glycosylation in addition to glycosyltransferase expression are operative in HEL cells and that they are part of a multi-regulated process controlling terminal modifications of glycoproteins.

Original languageEnglish (US)
Pages (from-to)159-166
Number of pages8
JournalEuropean Journal of Biochemistry
Volume225
Issue number1
DOIs
StatePublished - 1994
Externally publishedYes

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Leukemia, Erythroblastic, Acute
Glycoproteins
Cells
Cell Line
Glycosylation
Glycosyltransferases
galactoside 3-fucosyltransferase
Guanosine Diphosphate Fucose
alpha-L-Fucosidase
ABO Blood-Group System
Fucose
Biosynthesis
Enzymes
GDP-fucose-galactosyl-1-4-N-acetylglucosamine (Fuc to GlcNAc) alpha-1,3-fucosyltransferase
N-acetylglucopyranosylamine
Western Blotting
Antibodies
Substrates

ASJC Scopus subject areas

  • Biochemistry

Cite this

Expression of GDP-L-Fuc : Gal(β1-4)GlcNAc-R (Fuc to GlcNAc) α-1,3-fucosyltransferase and its relationship to glycoprotein structure in a human erythroleukemia cell line, HEL. / Giuntoli, R. L.; Stoykova, L. I.; Gillies, D. R B; Glick, M. C.

In: European Journal of Biochemistry, Vol. 225, No. 1, 1994, p. 159-166.

Research output: Contribution to journalArticle

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abstract = "Terminal glycosylation may be a mechanism to control the function of specific biologically active glycoproteins. The biosynthesis of terminal sialyl and fucosyl residues on certain glycoproteins has been linked to the expression of the respective glycosyltransferase. In contrast, a human erythroleukemia cell line, HEL, contained a highly active GDP-L-Fuc: Gal(β1-4)GlcNAc-R (Fuc to GlcNAc) α-1,3-fucosyltransferase (α-1,3-fucosyltransferase) but no detectable α-1,3-linked fucosyI residues on the glycoproteins. The α-1,3-fucosyltransferase gave apparent K(m) values for Fuc(α1-2)Gal(β1-4)GlcNAc β-O-benzyl, Gal(β1-4)GlcNAc and GDP-fucose of 0.04, 0.68 and 0.12 mM, respectively. The lack of detectable fucosyl residues in α-1,3-linkage to GlcNAc on the [3H]fucose-labeled glycoproteins was shown with the use of almond α-1,3/4-fucosidase and internal controls to verify that the enzyme was active. Using Western-blot analysis, HEL cell glycoproteins reacted with blood group H type-2 antibody, confirming the presence of Fuc(α1-2)Gal(β1-4)GlcNAc as reported by others and the presence of the preferred substrate for the enzyme. It is proposed that controls for terminal glycosylation in addition to glycosyltransferase expression are operative in HEL cells and that they are part of a multi-regulated process controlling terminal modifications of glycoproteins.",
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