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Blood, 15 November 2000, Vol. 96, No. 10, pp. 3480-3489
HEMOSTASIS, THROMBOSIS, AND VASCULAR BIOLOGY
Cytoskeletal regulation of the platelet glycoprotein Ib/V/IX-von
Willebrand factor interaction
Nayna Mistry,
Susan L. Cranmer,
Yuping Yuan,
Pierre Mangin,
Sacha M. Dopheide,
Ian Harper,
Simon Giuliano,
Dave E. Dunstan,
Francois Lanza,
Hatem H. Salem, and
Shaun P. Jackson
From the Department of Medicine, Australian Centre for
Blood Diseases, Monash Medical School, Victoria, Australia; Department
of Chemical Engineering, University of Melbourne, Victoria, Australia;
Etablissement de Transfusion Sanguine, Institut National de la Sante et
de la Recherche Medicale Unite 311, Strasbourg Cedex, France.
Shear-induced binding of von Willebrand factor (vWf) to the
platelet glycoprotein (GP) Ib/V/IX complex plays a key role in initiating platelet adhesion and aggregation at sites of vascular injury. This study demonstrated that pretreating human platelets with
inhibitors of actin polymerization, cytochalasin D or latrunculin B,
dramatically enhances platelet aggregation induced by vWf. The effects
of these inhibitors were specific to the vWf-GPIb interaction
because they enhanced vWf-induced aggregation of Glanzmann thrombasthenic platelets and Chinese hamster ovary (CHO) cells transfected with GPIb/V/IX. Moreover, cytochalasin D enhanced the
extent of platelet aggregation induced by high shear stress (5000 s 1) and also lowered the shear threshold required
to induce aggregation from 3000 s1 to as low as 500 s1. Studies of CHO cells expressing GPIb cytoplasmic
tail truncation mutants that failed to bind actin-binding protein-280
(deletion of residues 569-610 or 535-568) demonstrated that the
linkage between GPIb and actin-binding protein-280 was not required for vWf-induced actin polymerization, but was critical for the enhancing effects of cytochalasin D on vWf-induced cell aggregation. Taken together, these studies suggest a fundamentally important role for the
cytoskeleton in regulating the adhesive function of
GPIb/V/IX.
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