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Characterization of the N-glycosylation phenotype of erythrocyte membrane proteins in congenital dyserythropoietic anemia type II (CDA II/HEMPAS)

Glycoconjugate Journal volume 25, pages 375–382 (2008)Cite this article

Abstract

Congenital dyserythropoetic anemia type II (CDA II) is characterized by bi- and multinucleated erythroblasts and an impaired N-glycosylation of erythrocyte membrane proteins. Several enzyme defects have been proposed to cause CDA II based on the investigation of erythrocyte membrane glycans pinpointing to defects of early Golgi processing steps. Hitherto no molecular defect could be elucidated. In the present study, N-glycosylation of erythrocyte membrane proteins of CDA II patients and controls was investigated by SDS-Page, lectin binding studies, and MALDI-TOF/MS mapping in order to allow an embracing view on the glycosylation defect in CDA II. Decreased binding of tomato lectin was a consistent finding in all typical CDA II patients. New insights into tomato lectin binding properties were found indicating that branched polylactosamines are the main target. The binding of Aleuria aurantia, a lectin preferentially binding to α1-6 core-fucose, was reduced in western blots of CDA II erythrocyte membranes. MALDI-TOF analysis of band 3 derived N-glycans revealed a broad spectrum of truncated structures showing the presence of high mannose and hybrid glycans and mainly a strong decrease of large N-glycans suggesting impairment of cis, medial and trans Golgi processing. Conclusion: Truncation of N-glycans is a consistent finding in CDA II erythrocytes indicating the diagnostic value of tomato-lectin studies. However, structural data of erythrocyte N-glycans implicate that CDA II is not a distinct glycosylation disorder but caused by a defect disturbing Golgi processing in erythroblasts.

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Acknowledgements

J.D. was supported by a grant of Innovative Medizinische Forschung (De 110030) and J.D. and T.M. by a grant of the Bundesministerium für Bildung und Forschung (BMBF 01GM0313). G.Kreissel, V.Sablitzky, and K.Wardecki are acknowledged for excellent technical assistance.

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Affiliations

  1. Department of Pediatrics, University Hospital of Münster, Albert-Schweitzer-Str. 33, 48149, Munster, Germany

    Jonas Denecke, Christian Kranz & Thorsten Marquardt

  2. Department of Hematology and Oncology, University Hospital of Ulm, Ulm, Germany

    Hermann Heimpel

  3. German Research Center for Biotechnology (GBF), GlycoThera inc, Braunschweig, Germany

    Manfred Nimtz & Harald S. Conradt

  4. Department of Pediatrics, University Hospital of Magdeburg, Magdeburg, Germany

    Thomas Brune

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  1. Jonas Denecke
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  2. Christian Kranz
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  3. Manfred Nimtz
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  4. Harald S. Conradt
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  5. Thomas Brune
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  6. Hermann Heimpel
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Correspondence to Jonas Denecke.

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Denecke, J., Kranz, C., Nimtz, M. et al. Characterization of the N-glycosylation phenotype of erythrocyte membrane proteins in congenital dyserythropoietic anemia type II (CDA II/HEMPAS). Glycoconj J 25, 375–382 (2008). https://doi.org/10.1007/s10719-007-9089-1

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  • DOI: https://doi.org/10.1007/s10719-007-9089-1

Keywords

  • Hempas
  • CDA II
  • Congenital disorders of glycosylation
  • Dyserythropoesis
  • Erythrocyte membrane glycosylation