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Published on: June 5, 2019
Platelet activation by dimeric beta2-glycoprotein I requires signaling via both glycoprotein Ibalpha and
R T Urbanus1, M T T Pennings, R H W M Derksen
1Department of Clinical Chemistry and Haematology, University Medical Centre Utrecht, Utrecht, The Netherlands.
Background:
Dimerization of beta(2)-glycoprotein I (beta(2)-GPI) by autoantibodies is thought to trigger the clinical manifestations observed in the antiphospholipid syndrome. Arterial thrombosis, a frequently occurring clinical manifestation of the antiphospholipid syndrome, is a process in which platelets play a crucial role. Previous work has shown that binding of dimeric beta(2)-GPI to the platelet receptors apolipoprotein E receptor 2' (ApoER2') and glycoprotein Ibalpha (GPIbalpha) mediates increased platelet activation in an in vitro thrombosis model.
Objective:
The individual roles of ApoER2' and GPIbalpha in mediating platelet activation by dimeric beta(2)-GPI has hitherto been unclear. In this study, we have determined the roles of either receptor in platelet activation by dimeric beta(2)-GPI.
Methods:
Platelet activation by dimeric beta(2)-GPI was studied under conditions of flow. Intracellular signaling induced by dimeric beta(2)-GPI was subsequently analyzed by means of sodium dodecylsulfate polyacrylamide gel electrophoresis (SDS-PAGE) and western blot analysis.
Results:
The increase in platelet deposition onto a fibronectin surface under conditions of flow by dimeric beta(2)-GPI was completely abolished by inhibition of the interaction of dimeric beta(2)-GPI with either GPIbalpha or ApoER2'. Upon platelet stimulation with dimeric beta(2)-GPI, GPIbalpha translocated to the cytoskeleton via the scaffold protein 14-3-3zeta. Concomitantly, ApoER2' dissociated from the adapter protein Disabled1, presumably through phosphorylation of the cytoplasmic tail. Inhibition of one process could not inhibit the other.
Conclusion:
We show that dimeric beta(2)-GPI signals via two distinct pathways in platelets, both of which are required for platelet activation. Abrogation of either signal results in loss of activation.
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