Platelet protein S directly inhibits procoagulant activity on platelets and microparticles

Fabian Stavenuiter; Nicole F. Davis; Erning Duan; Andrew J. Gale; Mary J. Heeb

doi:10.1160/TH12-08-0622

Thrombosis and Haemostasis, Table of Contents

Thromb Haemost 2013; 109(02): 229-237
DOI: 10.1160/TH12-08-0622

Blood Coagulation, Fibrinolysis and Cellular Haemostasis

Schattauer GmbH

Platelet protein S directly inhibits procoagulant activity on platelets and microparticles^[*]

Authors

Fabian Stavenuiter

¹Department of Molecular and Experimental Medicine, The Scripps Research Institute, La Jolla, California, USA
Nicole F. Davis

¹Department of Molecular and Experimental Medicine, The Scripps Research Institute, La Jolla, California, USA
Erning Duan

¹Department of Molecular and Experimental Medicine, The Scripps Research Institute, La Jolla, California, USA
Andrew J. Gale

¹Department of Molecular and Experimental Medicine, The Scripps Research Institute, La Jolla, California, USA
Mary J. Heeb

¹Department of Molecular and Experimental Medicine, The Scripps Research Institute, La Jolla, California, USA

Abstract

Summary

Anticoagulant plasma protein S (PS) is essential for maintaining haemostatic balance. About 2.5% of PS is stored in platelets and released upon platelet stimulation. So far, little is known about the functionality and importance of platelet (plt)PS. A platelet-associated protease cleaves plasma-derived (pd)PS and pltPS in the “thrombin-sensitive region”, abolishing activated protein C (APC) cofactor activity. However, we showed that cleaved PS retains APC-independent anticoagulant activities (“PS-direct”). To investigate whether pltPS or pdPS exert PS-direct on platelets or platelet-shed microparticles, thrombin and factor (F)Xa generation on unstimulated or stimulated washed platelets and microparticles were measured. Western blotting revealed that pltPS and pdPS bound to washed, stimulated platelets and microparticles, and that pltPS had slower electrophoretic mobility than pdPS. Platelet stimulation in the presence of inhibitory anti-PS antibodies resulted in 2.6 ± 1.6-fold (p<0.0004, n=20) more thrombin generation upon addition of FXa and prothrombin. PltPS exerted PSdirect that was similar to or greater than that of Zn²⁺-containing pdPS and much greater than that of Zn²⁺-deficient pdPS. Findings were confirmed using purified pltPS. Platelet-bound pltPS and microparticlebound pltPS had similar PS-direct. Finally, platelet stimulation in the presence of inhibitory anti-PS antibodies resulted in 1.5 ± 0.2-fold (p<0.0001, n=11) more FXa generation upon addition of TF/FVIIa and FX. Thus, pltPS inhibits both prothrombinase and extrinsic FXase activities. Neutralising antibodies against APC and TFPI had no effect on the PS-direct of pltPS or pdPS on platelets. This study indicates that pltPS may be an essential pool of PS that counterbalances procoagulant activities on platelets.

Keywords

Activated protein C - protein S - platelets - microparticles - anticoagulant activity

Full Text

References

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Platelet protein S directly inhibits procoagulant activity on platelets and microparticles[*]

Authors

Platelet protein S directly inhibits procoagulant activity on platelets and microparticles^[*]