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Thromb Haemost 2002; 88(01): 104-110
DOI: 10.1055/s-0037-1613161
Review Article
Schattauer GmbH

Two Different β3 Cysteine Substitutions Alter αIIbβ3 Maturation and Result in Glanzmann Thrombasthenia

S. Milet-Marsal
1   UMR 5533 CNRS, Hôpital Cardiologique, Pessac, France
,
C. Breillat
1   UMR 5533 CNRS, Hôpital Cardiologique, Pessac, France
,
O. Peyruchaud
1   UMR 5533 CNRS, Hôpital Cardiologique, Pessac, France
,
P. Nurden
1   UMR 5533 CNRS, Hôpital Cardiologique, Pessac, France
,
R. Combrié
1   UMR 5533 CNRS, Hôpital Cardiologique, Pessac, France
,
A. Nurden
1   UMR 5533 CNRS, Hôpital Cardiologique, Pessac, France
,
F. Bourre
1   UMR 5533 CNRS, Hôpital Cardiologique, Pessac, France
› Author AffiliationsThis work was supported by the Ligue Régionale d’Aquitaine contre le Cancer. S. Milet has a grant from the Ligue Nationale Française contre le Cancer.
Further Information

Publication History

Received 15 October 2001

Accepted after resubmission 25 March 2002

Publication Date:
09 December 2017 (online)

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Summary

We report the defects responsible for Glanzmann thrombasthenia in two patients showing traces of abnormally migrating platelet β3 in immunoblotting. Using PCR-SSCP and direct sequencing, we identified a novel homozygous mutation in exon 10 of the β3 gene of patient 1 which gave a C457 to Y amino acid substitution. A C542 to R substitution in β3 of patient 2 was previously reported by us. These cysteines are present in EGF-domains 1 and 3 respectively of β3. We therefore constructed mutants carrying substitutions on cysteine residues in each of the first three EGF domains of β3, C457, C495 and C542 respectively. Transient expression of these mutants in COS-7 cells, including the C542 and C547 double mutant, proved that disulfide disruption directly affects cell surface expression of the integrin. We then showed by metabolic (35S) labeling and Endo-H glycosidase treatment that these substitutions strongly affected complex maturation within the cell.

Keywords

Glanzmann thrombasthenia - platelet - αIIbβ3 integrin - cysteine mutations - Cos-7 cells
 

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