A novel homozygous mutation (1619delC) in GPIIb gene associated with Glanzmann thrombasthenia, the decay of GPIIb-mRNA and the synthesis of a truncated GPIIb unable to form complex with GPIIIa

Gergely Losonczy; Nurit Rosenberg; Csongor Kiss; János Kappelmayer; György Vereb; Adrienne Kerényi; István Balogh; László Muszbek

doi:10.1160/TH04-12-0848

Thrombosis and Haemostasis, Table of Contents

Thromb Haemost 2005; 93(05): 904-909
DOI: 10.1160/TH04-12-0848

Platelets and Blood Cells

Schattauer GmbH

A novel homozygous mutation (1619delC) in GPIIb gene associated with Glanzmann thrombasthenia, the decay of GPIIb-mRNA and the synthesis of a truncated GPIIb unable to form complex with GPIIIa

Gergely Losonczy

¹Clinical Research Center, University of Debrecen, Medical and Health Science Center, Debrecen, Hungary

,

Nurit Rosenberg

²Amalia Biron Research Institute of Thrombosis and Hemostasis, The Chaim Sheba Medical Center, Tel-Hashomer, Israel

,

Csongor Kiss

³Department of Pediatrics, University of Debrecen, Medical and Health Science Center, Debrecen, Hungary

,

János Kappelmayer

⁴Department of Clinical Biochemistry and Molecular Pathology, University of Debrecen, Medical and Health Science Center, Debrecen, Hungary

,

György Vereb

⁵Department of Biophysics and Cell Biology, University of Debrecen, Medical and Health Science Center, Debrecen, Hungary

,

Adrienne Kerényi

⁴Department of Clinical Biochemistry and Molecular Pathology, University of Debrecen, Medical and Health Science Center, Debrecen, Hungary

,

István Balogh

⁴Department of Clinical Biochemistry and Molecular Pathology, University of Debrecen, Medical and Health Science Center, Debrecen, Hungary

,

László Muszbek

¹Clinical Research Center, University of Debrecen, Medical and Health Science Center, Debrecen, Hungary

⁶Thrombosis and Haemostasis Research Group of the Hungarian Academy of Sciences, University of Debrecen, Medical and Health Science Center, Debrecen, Hungary

› Author Affiliations

Abstract

Summary

The absence of agonist-induced platelet aggregation and the lack of fibrinogen receptor (GPIIb/IIIa) on the platelet surface demonstrated that the severe hemorrhagic complications of a child of Romany descent were caused by Glanzmann thrombasthenia. DNA sequencing revealed a novel homozygous deletion of a cytosine (1619delC) in the GPIIb gene causing a frameshift and predicting a novel stop codon at position 533 following 24 altered amino acids. Both parents possessed the same deletion in heterozygous form. The amount of GPIIb mRNA in the patient’s platelets was 0.06% of the amount measured in control platelets. Neither GPIIb nor its truncated form could be detected in the platelets of the patient by Western blotting, while a small amount of GPIIIa was demonstrated. Quantitative flow cytometric analysis showed an elevated number of vitronectin receptors, a component of which is GPIIIa, on the patient’s platelets. The surface expression of vitronectin receptor on thrombasthenic, but not on normal platelets was further increased by activation with thrombin receptor agonist peptide. BHK cells transfected with wild type GPIIIa and mutated GPIIb failed to express any mature GPIIb or pro-GPIIb. Immunoprecipitation with a polyclonal antibody recognizing both GPIIb and GPIIIa recovered a 60 kDa truncated form of GPIIb. This band was absent when immunoprecipitation was carried out with an antibody recognizing GPIIIa, suggesting that the truncated protein, lacking calf-1, calf-2 domains and major part of the thigh domain, is unable to form complex with GPIIIa.

Keywords

Fibrinogen receptor - Glanzmann thrombasthenia - glycoprotein IIb mutation - truncated glycoprotein IIb - vitronectin receptor

Full Text

References

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