Abstract
One of the biochemical characteristics of carbohydrate deficient glycoprotein syndromes is the presence of abnormal glycoforms in serum transferrin. Both glycoform heterogeneity and variable site occupancy may, in principle, lead to the generation of a range of glycoforms which contain different numbers of sialic acid residues, and therefore variable amounts of negative charge. Capillary zone electrophoresis was used to resolve the glycoforms of normal human serum transferrin and also of a set of glycoforms which were prepared by digesting the sugars on the intact glycoprotein with sialidase. The sugars on the intact glycoprotein were also modified by a series of exoglycosidase enzymes to produce a series of neutral glycoforms which were also analysed by capillary zone electrophoresis. The oligosaccharide population of human serum transferrin was analysed by a series of mixed exoglycosidase digests on the released glycan pool and quantified using a novel HPLC strategy. Transferrin was isolated from carbohydrate deficient glycoprotein syndromes type I serum and both the intact glycoforms and released sugars were resolved and quantified. The data presented here confirm the presence of a hexa-, penta- and tetra-sialoforms of human serum transferrin in both normal and carbohydrate deficient glycoprotein syndrome type I serum samples. Consistent with previous reports carbohydrate deficient glycoprotein syndrome type I transferrin also contained a di-sialoform, representing a glycoform in which one of the two N-glycosylation sites is unoccupied, and a non-glycosylated form where both remain unoccupied. This study demonstrates that capillary zone electrophoresis can be used to resolve quantitatively both sialylated and neutral complex type glycoforms, suggesting a rapid diagnostic test for the carbohydrate deficient glycoprotein syndromes group of diseases.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
Abbreviations
- CDGS:
-
Carbohydrate Deficient Glycoprotein Syndrome
- CZE:
-
Capillary Zone Electrophoresis
- hTf:
-
human transferrin
- gu:
-
HPLC glucose units
- EOF:
-
electroosmotic flow. Nomenclature: for describing oligosaccharide structures: A(1,2,3,4) indicates the number of antennae linked to the t trimannosyl core
- G(0–4):
-
indicates the number of terminal galactose residues in the structure
- F:
-
core fucose
- B:
-
bisecting N-acetyl glucosamine (GlcNAc)
- S:
-
sialic acid
- Gal:
-
galactose; M
- Man:
-
mannose
References
Spik G, Bayard B, Fournet B, Strecker G, Bouqualt S, Montreuil J (1975)FEBS Lett 50: 296–9.
Dorland L, Haverkamp J, Schut BL, Vliegenhart JFG, Spik G, Strecker G, Fournet B, Montreuil J (1977)FEBS Lett 77: 15–206.
Fu D, van Halbeek H (1992)Anal Biochem 206: 53–206.
De Jong G, Van Noort WL, Feelders RA, de Jeu Jaspers CMH, van Eijk HG (1992)Clin Chim Acta 212: 27–45.
Spik G, Coddeville B, Montreuil J (1988)Biochemie 70: 1459–69.
Hershberger CL, Larson JL, Arnold B, Rosteck PR, Williams P, DeHoff B, Dunn P, O'Neal KL, Riemen MW, Tice PA, Cofts R, Ivancic J (1991)Ann NY Acad Sci 646: 140–54.
Mason AB, Miller MK, Funk WD, Banfield DK, Savage KJ, Oliver RWA, Green BN, MacGillivray RTA, Woodworth RC (1993)Biochemistry 32: 5472–9.
Stibler H, Borg S, Joustra M (1986)Alcoholism: Clinical and Experimental Research 10: 535–44.
Landberg E, Pahlsson P, Lundblad A, Arnetorp A, Jeppsson JO (1995)Biochem Biophys Res Commun. 210: 267–742.
Jaeken J, van Eijk HG, van der Heul C, Corbeel L, Eeckels R, Eggermont E (1984)Clin Chim Acta 144: 245–7.
Jaeken J, Carchon H, Stibler H (1993)Glycobiology 3: 423–8.
Jaeken J, Schacter H, Carchon H, De Cock P, Coddeville B, Spik G (1994)Arch Dis Childhood,71: 123–7.
Stibler H, Westerberg B, Hanefield F, Hagberg B (1992)Neuropediatrics 24: 51–2.
Stibler H, Stephani U, Kutsch U (1995)Neuropediatrics 26: 235–7.
Yamashita K, Ideo H, Ohkura T, Fukushima K, Yusa I, Ohno K, Takeshita K (1993)J Biol Chem 268: 5783–9.
Wada Y, Nishikawa A, Okamoto N, Inui K, Tsukamoto H, Okada S, Taniguchi N (1992)Biochem Biophys Res Commun 189: 832–68.
Marquardt T, Ullrich K, Zimmer P, Hasilik A, Denfel T, Harms E (1995)Europ J Cell Biol 86: 268–73.
Powell LD, Paneerselvan K, Vij R, Diaz S, Manzi A, Baist N, Freeze H, Varki A (1994)J Clin Invest 94: 1901–9.
Van Schaftingen E, Jaeken J (1995)FEBS Lett 337: 318–20.
Martinsson T, Bjursell, Stibler H, Kristiansson B, Skonsby F, Jaeken J, Blennow G, Stromme P, Hanefeld F, Wahlstrom J (1994)Human Mol Genetics 3: 2037–42.
Kilar F, Hjerten S (1989)Electrophoresis 10: 23–9.
van Eijk HG, van Noort WL (1992)Electrophoresis 13: 354–8.
Karger BL, Chu YH, Foret F (1995)Ann Rev Biophys-Biomol Structure 24: 579–610.
Rudd PM, Scragg IG, Coghill E, Dwek RA (1992)Glyco. J. 9: 86–91.
Steinbuch N, Niewiarowski S (1960)Nature 186: 87–8.
Guile GR, Rudd PM, Wing DR, Prime FB, Dwek RA (1996)Anal. Biochem 240: 210–226.
Yamashita K, Koide N, Endo T, Iwaki Y, Kobata A (1989)J Biol Chem 264: 2415–23.
Jacob GS, Scudder P (1994)Meth Enzymol 230: 280–99.
Edge CJ, Rademacher TW, Wormald MR, Parekh RB, Butters TD, Wing DR, Dwek RA (1992)Proc Natl Acad Sci 89: 6338–42.
Rudd PM, Joao HC, Coghill E, Fiten P, Saunders MR, Opdenakker G, Dwek RA (1994)Biochemistry 33: 17–22.
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Iourin, O., Mattu, T.S., Mian, N. et al. The identification of abnormal glycoforms of serum transferrin in carbohydrate deficient glycoprotein syndrome type i by capillary zone electrophoresis. Glycoconjugate J 13, 1031–1042 (1996). https://doi.org/10.1007/BF01053199
Received:
Revised:
Accepted:
Issue Date:
DOI: https://doi.org/10.1007/BF01053199