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Thromb Haemost 1999; 81(05): 782-792
DOI: 10.1055/s-0037-1614571
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Function of Glycoprotein VI and Integrin α2β1 in the Procoagulant Response of Single, Collagen-Adherent Platelets

Johan W. M. Heemskerk
1   From the Departments of Biochemistry and Human Biology, University of Maastricht, Maastricht, The Netherlands; the
,
Pia Siljander
2   Wihuri Research Institute, Helsinki, Finland; University of Cambridge, Cambridge, United Kingdom
,
Wim M. J. Vuist
1   From the Departments of Biochemistry and Human Biology, University of Maastricht, Maastricht, The Netherlands; the
,
Githa Breikers
1   From the Departments of Biochemistry and Human Biology, University of Maastricht, Maastricht, The Netherlands; the
,
Chris P. M. Reutelingsperger
1   From the Departments of Biochemistry and Human Biology, University of Maastricht, Maastricht, The Netherlands; the
,
Michael J. Barnes
3   Department of Biochemistry, University of Cambridge, Cambridge, United Kingdom
,
C. Graham Knight
3   Department of Biochemistry, University of Cambridge, Cambridge, United Kingdom
,
Riitta Lassila
2   Wihuri Research Institute, Helsinki, Finland; University of Cambridge, Cambridge, United Kingdom
,
Richard W. Farndale
3   Department of Biochemistry, University of Cambridge, Cambridge, United Kingdom
› Author Affiliations
Further Information

Publication History

Received 22 September 1998

Accepted after revision 22 January 1999

Publication Date:
09 December 2017 (online)

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Summary

Various collagen-based materials were used to assess the structural requirements of collagen for inducing the procoagulant response of adhering platelets, as well as the collagen receptors involved. Cross-linked or monomeric collagen-related peptide (CRP), Gly-Cys-Hyp-(Gly-Pro-Hyp)10-Gly-Cys-Hyp-Gly was highly adhesive for platelets in a glycoprotein VI- (GpVI-)dependent manner. Adhesion was followed by a prolonged increase in cytosolic [Ca2+]i, formation of membrane blebs, exposure of phosphatidylserine (PS) and generation of prothrombinase-stimulating activity. Fibrils of type-I collagen were less adhesive but, once adhered, many of the platelets presented a full procoagulant response. Monomeric type-I collagen was unable to support adhesion, unless Mg2+-dependent integrin α2β1 interactions were facilitated by omission of Ca2+ ions. With all surfaces, however, post-addition of CaCl2 to adhering platelets resulted in a potent Ca2+-influx signal, followed by PS exposure and bleb formation. The procoagulant response elicited by binding to CRP was inhibited by anti-GpVI Fab fragments, but not by impeding integrin α2β1-mediated events. With fibrillar collagen, it was inhibited by blocking either the GpVI- or integrin α2β1- mediated interactions. This suggests that the triple-helical Gly-Pro-Hyp repeat in CRP and analogous sequences in fibrillar collagen stimulate the procoagulant response of adherent platelets by acting as ligands for GpVI. Influx of Ca2+ is required for this response, and adhesion via integrin α2β1 serves to potentiate the signaling effects of GpVI.

 

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