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[*]

Fabian Stavenuiter
1   Department of Molecular and Experimental Medicine, The Scripps Research Institute, La Jolla, California, USA
,
Nicole F. Davis
1   Department of Molecular and Experimental Medicine, The Scripps Research Institute, La Jolla, California, USA
,
Erning Duan
1   Department of Molecular and Experimental Medicine, The Scripps Research Institute, La Jolla, California, USA
,
Andrew J. Gale
1   Department of Molecular and Experimental Medicine, The Scripps Research Institute, La Jolla, California, USA
,
Mary J. Heeb
1   Department of Molecular and Experimental Medicine, The Scripps Research Institute, La Jolla, California, USA
› Author Affiliations
Financial support:This work was partly supported by National Institutes of Health grant HL088375.
Further Information

Publication History

Received: 28 August 2012

Accepted after minor revision: 09 November 2012

Publication Date:
15 December 2017 (online)

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.

* A preliminary report of this study was presented in abstract form at the American Society of Hematology annual meeting, December, 2010.


 
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