Glycophorin A in two patients with congenital dyserythropoietic anemia type I and type II is partly unglycosylated.

• Authors: Ewa Zdebska , Monika Adamczyk-Popławska , Jerzy Kościelak
• Languages of publication: EN

Abstracts

EN

Glycophorins A from erythrocyte membranes of two patients with congenital dyserythropoietic anemia type I and type II (CDA type I and II) were analyzed for carbohydrate molar composition employing a modification of the recently published method that allowed simultaneous determination of carbohydrates and protein in electrophoretic bands of glycoproteins separated by sodium dodecyl sulfate- polyacrylamide gel electrophoresis (Zdebska & Kościelak, 1999, Anal. Biochem., 275, 171-179). The modification involved a preliminary extraction of erythrocyte membranes with aqueous phenol, subsequent electrophoresis and analysis of the extracted glycophorins rather than electrophoresis and analysis of the glycophorin from intact erythrocyte membranes. The results showed a large deficit of N-acetylgalactosamine, galactose, and sialic acid residues in glycophorin A from patients with CDA type I and type II amounting to about 45% and 55 %, respectively. The results strongly suggest that glycophorin A in these patients is partly unglycosylated with respect to O-linked glycans. In addition, glycophorin A from erythrocytes of a patient with CDA II but not CDA I exhibited a significant deficit of mannose and N-acetylglucosamine suggesting that its N-glycosylation site was also partly unglycosylated.

Keywords

EN

congenital dyserythropoietic anemia type I and II; unglycosylation; glycophorin A

• Publisher: Polish Biochemical Society
• Journal: Acta Biochimica Polonica
• Year: 2000
• Volume: 47
• Issue: 3
• Pages: 773-779

Dates

published

2000

received

2000-07-18

accepted

2000-07-27

Contributors

author

Ewa Zdebska

• Department of Biochemistry, Institute of Hematology and Blood Transfusion, Warszawa, Poland

author

Monika Adamczyk-Popławska

• Department of Biochemistry, Institute of Hematology and Blood Transfusion, Warszawa, Poland

author

Jerzy Kościelak

• Department of Biochemistry, Institute of Hematology and Blood Transfusion, Warszawa, Poland

References

• Aebi, M., Barone, R., Berger, E.G., Bjursell, C., Briones, P., Cardo, E., Clayton, P., Cormier-Daire, V., Cuer, M., de Koning, T., de Lonay, P., de Loos, F., Dorland, L., Dupre, T., Fabritz, L., Fiumara, A., Freeze, H., Grunewald, S., Hasilik, M., Helenius, A., Hennet, T., Heykants, L., Imbach, T., Jaeken, J., Keir, G., Kjaergaard, S., Klein, A., Kościelak, J., Körner, C., Krasnewich, D., Kupers, L., Lehle, L., Lubke, T., Marquardt, T., Matthijs, G., Niehues, R., Peters, V., Raab, Roussel, P.M., Saether, O., Seta, N., Schachter, H., Schenk, B., Schollen, E., Schwartz, M., Skovby, F., Stutz, A., Tamashita, K., Thiel, C., Uller, A., van Schaftingen, E., Verbert, A., von Figura, K., Vilaseca, A., Wahlstrom, J.G., Wevers, R. & Winchester, B. (2000) Carbohydrate-deficient glycoprotein syndromes become congenital disorders of glycosylation: An updated nomenclature for CDG. Glycobiology 10, Glycoconjugate J., 16 (in press).
• Bruce, L.J., Groves, J.D., Okubo, Y., Thilaganathan, B. & Tanner, M.J.A. (1994) Altered band 3 structure and function in glycophorin A- and B-deficient (MkMk) red blood cells. Blood 84, 916-922.
• Carchon, H., Van Schaftingen, E., Matthijs, G. & Jaeken, J. (1999) Carbohydrate-deficient glycoprotein syndrome type IA (phosphomannomutase-deficiency). Biochim. Biophys. Acta 1455, 155-165.
• Crookston, J.H., Crookston, M.C., Brunie, K.L., Francombe, W.H., Dacie, J.V., Davis, J.A. & Lewis, S.M. (1969) Hereditary erythroblastic multinuclearity associated with a positive acidified-serum test: A type of congenital dyserythropoietic anaemia. Br. J. Haematol. 17, 11-26.
• Dahr, W., Moulds, J., Unger, P. & Kordowicz, M. (1987) The Dantu erythrocyte phenotype of the NE variety. I. Dodecylsulfate polyacrylamide gel electrophoresis studies. Blut 55, 19- 31.
• Daniels, G. (1999) Fuctional aspects of red cell antigens. Blood Rev. 13, 14-35.
• Dodge, J.T., Mitchell, C. & Hanahan, D.J. (1963) The preparation and chemical characteristics of hemoglobin-free ghosts of human erythrocytes. Arch. Biochem. Biophys. 100, 119- 130.
• Fukuda, M.N. (1990) HEMPAS disease: Genetic defect of glycosylation. Glycobiology 1, 9-15.
• Fukuda, M.N. (1999) HEMPAS. Biochim. Biophys. Acta 1455, 231-239.
• Gahmberg, C.G., Myllyla, G., Leikola, J., Pirkola, A. & Nordling, S. (1976) Absence of the major sialoglycoprotein in the membrane of human En(a-) erythrocytes and increased glycosylation of band 3. J. Biol. Chem. 251, 6108- 6116.
• Gasparini, P., del Giudice, E.M., Delaunay, J., Totaro, A., Granatiero, M., Merchionda, S., Zalante, L. & Iolascon, A. (1997) Localization of the congenital dyserythropoietic anemia II locus to chromosome 20q11.2 by genomewide search. Am. J. Hum. Genet. 61, 1112-1116.
• Hassoun, H., Hanada, T. & Lutchman, M (1998) Complete deficiency of glycophorin A in red blood cells from mice with targeted inactivation of the band 3 (AE1) gene. Blood 91, 2146-2151.
• Heimpel, H. & Wendt, F. (1968) Congenital dyserythropoietic anaemia with karyorrhexis and multinuclearity of erythroblasts. Helv. Med. Acta 34, 103-115.
• Higgins, T.J. (1984) Simplified fluorometric assay for sphingosine bases. J. Lipid Res. 25, 1007-1009.
• Iolascon, A., del Giodice, E.M., Perrotta, S., Granatiero, M., Zelante, L. & Gasparini, P. (1997) Exclusion of three candidate genes as determinants of congenital dyserythropoietic anemia type II (CDA-II). Blood 90, 4197-4200.
• Krotkiewska, B., Zwierz, K. & Krotkiewski, H. (1999) The carbohydrate moiety of human glycophorin in CDG syndrome. Acta Biochim. Polon. 46, 371-376.
• Krotkiewski, H., Duk, M., Syper, D., Lis, H., Sharon, N. & Lisowska, E. (1997) Blood group MN-dependent difference in degree of galactosylation of O-glycans of glycophorin A is restricted to the GalNAc residues located on amino acid residues 2-4 of the glycophorin polypeptide chain. FEBS Lett. 406, 296-300.
• Lisowska, E., Messeter, L., Duk, M., Czerwiński, M. & Lundblad, A. (1987) A monoclonal anti-glycophorin A antibody recognizing the blood M determinant: Studies on the subspecificity. Mol. Immunol. 24, 605-614.
• Mawby, W.J., Tanner, M.J.A., Anstee, D.J. & Clamp, J.R (1983) Incomplete glycosylation of erythrocyte membrane proteins in congenital dyserythropoietic anaemia type II (CDA II). Br. J. Haematol. 55, 357-368.
• Pisano, A., Redmond, J.W., Williams, K.R. & Gooley, A.A (1993) Glycosylation sites identified by solid-phase Edman degradation: O-linked glycosylation motifs on human glycophorin A. Glycobiology 3, 429-435.
• Poole, J. (2000) Red cell antigens on band 3 and glycophorin A. Blood Rev. 14, 31-43.
• Tanner, M.J.A., Jenkins, R.E., Anstee, D.J. & Clamp, J.R. (1976) Abnormal carbohydrate composition of the major membrane penetrating protein of En(a-) human erythrocytes. Biochem. J. 155, 701-703.
• Tomita, A. & Parker, Ch.J. (1994) Aberrant regulation of complement by the erythrocytes of hereditary erythroblastic multinuclearity with a positive acidified serum lysis test (HEMPAS). Blood 83, 250-259.
• Undefriend, S., Stein, S., Bohlen, P., Dairman, W., Leimbgruber, W. & Weigele, M. (1972) Fluorescamine: A reagent for assay of amino acids, peptides, proteins, and primary amines in the picomole range. Science 178, 871-872.
• Wickramasinghe, S.N. (1998) Congenital dyserythropoietic anaemias: Clinical features, haematological morphology an new biochemical data. Blood Rev. 12, 178-200.
• Wickramasinghe, S.N. (2000) Congenital dyserythropoietic anemias. Curr. Opin. Hematol. 7, 71-78.
• Zdebska, E. & Kościelak, J. (1999) A single-sample method for determination of carbohydrate and protein contents in glycoprotein bands separated by sodium dodecyl sulfate- polyacrylamide gel electrophoresis. Anal. Biochem. 275, 171-179.
• Zdebska, E., Jaeken, J. & Kościelak, J. (2000a) Band 3 glycoprotein and glycophorin A from erythrocytes of children with CDG Ia are underglycosylated. Electrophoresis (in press).
• Zdebska, E., WoŻniewicz, B., Adamowicz-Salach, A. & Kościelak, J. (2000b) Erythrocyte membranes from a patient with congenital dyserythropoietic anaemia type I (CDA-I) show identical, though less pronounced, glycoconjugate abnormalities to those from patients with CDA-II (HEMPAS). Br. J. Haematol. (in press).