A novel mutation in the transmembrane region of glycoprotein IX associated with Bernard-Soulier syndrome

Zhaoyue Wang; Xiaojuan Zhao; Weiming Duan; Jianxin Fu; Mingen Lu; Giamin Wang; Xia Bai; Changgeng Ruan

doi:10.1160/TH04-04-0240

Thrombosis and Haemostasis, Inhaltsverzeichnis

Thromb Haemost 2004; 92(03): 606-613
DOI: 10.1160/TH04-04-0240

Platelets and Blood Cells

Schattauer GmbH

A novel mutation in the transmembrane region of glycoprotein IX associated with Bernard-Soulier syndrome

Authors

Zhaoyue Wang

¹Jiangsu Institute of Hematology,The First Affiliated Hospital of Soochow University, China
Xiaojuan Zhao

¹Jiangsu Institute of Hematology,The First Affiliated Hospital of Soochow University, China
Weiming Duan

¹Jiangsu Institute of Hematology,The First Affiliated Hospital of Soochow University, China
Jianxin Fu

¹Jiangsu Institute of Hematology,The First Affiliated Hospital of Soochow University, China
Mingen Lu

¹Jiangsu Institute of Hematology,The First Affiliated Hospital of Soochow University, China
Giamin Wang

¹Jiangsu Institute of Hematology,The First Affiliated Hospital of Soochow University, China
Xia Bai

¹Jiangsu Institute of Hematology,The First Affiliated Hospital of Soochow University, China
Changgeng Ruan

¹Jiangsu Institute of Hematology,The First Affiliated Hospital of Soochow University, China

Abstract

Summary

We describe here a novel mutation in glycoprotein (GP) IX transmembrane region in a patient with Bernard-Soulier syndrome (BSS). Flow cytometric analysis of the patient’s platelets showed that GP Iba and GP IX were expressed at decreased levels. Sequence analysis of the gene coding for GP IX revealed a homozygous (G to A) transition at nucleotide 2113, resulting in a Ala 140 (GCC) to Thr (ACC) replacement in the mature peptide, whereas no defects were found in the coding region of the GP Iba and GP Ib? gene. Allele-specific restriction enzyme analysis using HPYCH4 III revealed that the patient was homozygous and her mother and brother were heterozygous for the defect, and excluded the possibility that the mutation was a polymorphism of GP IX.To clarify the effect of this mutation on the surface expression of the GP Ib/IX complex, we introduced this mutation into the cDNA of GP IX by site-directed mutagenesis and performed in vitro transfection studies with plasmids harboring GP Iba, GP Ib? and wild-type GP IX or mutant GP IX. Mutant GP IX decreased the surface expression of GP Iba and GP IX, whereas both immunostaining and immunoblotting of the transfected Chinese hamster ovary (CHO) cells showed abundant GP Iba and GP IX in the cytoplasm of the CHO cells transfected with plasmids harboring GP Iba, GP Ib? and wild-type GP IX or mutant GP IX These findings indicate that the Ala140→Thr mutation in the transmembrane region of GP IX does not induce intracellular GP Ib/IX complex degradation, but prevents its insertion in the cytoplasmic membrane of platelets and CHO cells.

Keywords

Bernard-Soulier syndrome - platelet - glycoprotein - gene - mutation

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Referenzen

References
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