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Thromb Haemost 2005; 93(04): 716-723
DOI: 10.1160/TH04-09-0584
Platelets and Blood Cells
Schattauer GmbH

Glycoprotein VI is associated with GPIb-IX-V on the membrane of resting and activated platelets

Jane F. Arthur
1   Department of Biochemistry and Molecular Biology, Monash University, Clayton, Victoria, Australia
,
Elizabeth E. Gardiner
1   Department of Biochemistry and Molecular Biology, Monash University, Clayton, Victoria, Australia
,
Maria Matzaris
1   Department of Biochemistry and Molecular Biology, Monash University, Clayton, Victoria, Australia
,
Simon G. Taylor
1   Department of Biochemistry and Molecular Biology, Monash University, Clayton, Victoria, Australia
,
Lakshmi Wijeyewickrema
1   Department of Biochemistry and Molecular Biology, Monash University, Clayton, Victoria, Australia
,
Yukio Ozaki
2   Department of Clinical and Laboratory Medicine, Faculty of Medicine, University of Yamanashi, Tamaho, Nakakoma, Yamanashi, Japan
,
Mark L. Kahn
3   Department of Medicine, University of Pennsylvania, Philadelphia, Pennsylvania, USA
,
Robert K. Andrews
1   Department of Biochemistry and Molecular Biology, Monash University, Clayton, Victoria, Australia
,
Michael C. Berndt
1   Department of Biochemistry and Molecular Biology, Monash University, Clayton, Victoria, Australia
› Author AffiliationsFinancial support: This study was partly supported by the National Health and Medical Research Council of Australia, and the National Heart Foundation of Australia.
Further Information

Publication History

Received 10 September 2004

Accepted after revision 27 January 2005

Publication Date:
14 December 2017 (online)

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Summary

The platelet collagen receptor, glycoprotein (GP)VI, initiates platelet aggregation at low shear stress while GPIb-IX-V, which binds von Willebrand factor, elicits platelet aggregation under high shear conditions. To investigate the possibility that GPIb-IX-V and GPVI are associated on the platelet surface, we first ascertained that aggregation induced by a GPVI-specific agonist, collagen-related peptide, like collagen, is markedly cross-blocked by a GPIbα-specific monoclonal antibody, SZ2. Immunoprecipitation of GPIb-IX with anti-GPIbα from the 1% (v/v) Triton-soluble fraction of unstimulated platelets and immunoblot-ting with anti-GPVI demonstrated association between GPIb-IX and GPVI. This association was maintained when platelets were activated by thrombin. Pre-treatment of platelets with methyl-β-cyclodextrin to disrupt lipid rafts did not affect association in resting platelets under these conditions of detergent lysis. The association is also independent of cytoskeletal attachment, since it was unaffected by treatment with N-ethylmaleimide or DNaseI, which dissociate GPIb-IX from filamin and the actin-containing cytoskeleton, respectively. Finally, the association involves an interaction between the ectodomains of GPIbα and GPVI, since soluble fragments of GPIbα (glycocalicin) and GPVI are co-precipitated from the platelet supernatant under conditions where GPVI is shed. A contribution of GPIb-IX-V to GPVI-induced platelet responses, and vice versa, therefore warrants further investigation.

Keywords

Platelet physiology - adhesion molecules - protein function/activity
 

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