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 language | English (US) |
|---|---|
| Pages (from-to) | 159-166 |
| Number of pages | 8 |
| Journal | European Journal of Biochemistry |
| Volume | 225 |
| Issue number | 1 |
| DOIs | |
| State | Published - 1994 |
| Externally published | Yes |
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- 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 journal › Article
}
TY - JOUR
T1 - Expression of GDP-L-Fuc
T2 - 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
AU - Giuntoli, R. L.
AU - Stoykova, L. I.
AU - Gillies, D. R B
AU - Glick, M. C.
PY - 1994
Y1 - 1994
N2 - 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.
AB - 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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UR - http://www.scopus.com/inward/citedby.url?scp=0027968480&partnerID=8YFLogxK
U2 - 10.1111/j.1432-1033.1994.00159.x
DO - 10.1111/j.1432-1033.1994.00159.x
M3 - Article
C2 - 7925433
AN - SCOPUS:0027968480
VL - 225
SP - 159
EP - 166
JO - FEBS Journal
JF - FEBS Journal
SN - 1742-464X
IS - 1
ER -