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Thromb Haemost 1996; 76(05): 768-773
DOI: 10.1055/s-0038-1650658
Original Article
Schattauer GmbH Stuttgart

Functional Expression of Single Chain Glycoprotein lb Alpha on the Surface of COS Cells and BHK Cells

Eefke J Petersen
The Department of Haematology, University Hospital Utrecht, The Netherlands
,
Eelke Posthumus
The Department of Haematology, University Hospital Utrecht, The Netherlands
,
Jan J Sixma
The Department of Haematology, University Hospital Utrecht, The Netherlands
› Author Affiliations
Further Information

Publication History

Received 06 January 1995

Accepted after resubmission 21 June 1996

Publication Date:
11 July 2018 (online)

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Summary

The GpIb-IX complex constitutes the major receptor for von Wille-brand factor (vWF) on the surface of blood platelets. The vWF-binding site has been mapped to one of the three constituent chains, Gplba. Surface expression of Gplba depends on the. correct intracellular assembly with the Gplb(3 and GpIX chains. We have now grafted a portion of the extracellular domain of Gplba containing the vWF binding site onto the transmembrane/intracellular domain of the single chain surface molecule ICAM-1. Transient transfection of this chimeric protein in COS cells resulted in surface expression as assessed by immunostain-ing of live cells. Similar results were obtained after stable transfection into BHK cells. Purified vWF bound to the surface of transfected cells in the presence of ristocetin and botrocetin with a Kd of 52 ng/ml, comparable to the Kd for fixed platelets (65.5 ng/ml). This study indicates that functional expression of the vWF-binding domain of GpIbα on the surface of mammalian cells can be obtained in the absence of GpIbβ and GpIX. Furthermore, this model system simplifies existing methods for the assessment of the functional consequences of mutations in Gplba as found in pseudo-von Willebrand disease and Bemard-Soulier syndrome.

 

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