Download PDF
Thromb Haemost 2012; 107(03): 468-476
DOI: 10.1160/TH11-07-0457
Blood Coagulation, Fibrinolysis and Cellular Haemostasis
Schattauer GmbH

Platelet-mediated proteolytic down regulation of the anticoagulant activity of protein S in individuals with haematological malignancies

Ilze Dienava-Verdoold
1   Department of Plasma Proteins, Sanquin Research, Amsterdam, The Netherlands
,
Marina R. Marchetti
2   Immunohematology and Transfusion Medicine Department, Ospedali Riuniti di Bergamo, Bergamo, Italy
,
Liane C. J. te Boome
3   Department of Hematology, Academical Medical Center, University of Amsterdam, Amsterdam, The Netherlands
4   Present address: Department of Hematology, Academic Medical Center, University of Utrecht, Utrecht, The Netherlands
,
Laura Russo
2   Immunohematology and Transfusion Medicine Department, Ospedali Riuniti di Bergamo, Bergamo, Italy
,
Anna Falanga
2   Immunohematology and Transfusion Medicine Department, Ospedali Riuniti di Bergamo, Bergamo, Italy
,
Harry R Koene
3   Department of Hematology, Academical Medical Center, University of Amsterdam, Amsterdam, The Netherlands
5   Pressent address: Department of Internal Medicine, St. Antonius Hospital, Nieuwegein, The Netherlands
,
Koen Mertens
1   Department of Plasma Proteins, Sanquin Research, Amsterdam, The Netherlands
6   Department of Pharmaceutics, Utrecht Institute for Pharmaceutical Sciences, Utrecht University, Utrecht, The Netherlands
,
Herm Jan M. Brinkman
1   Department of Plasma Proteins, Sanquin Research, Amsterdam, The Netherlands
› Author Affiliations
Further Information

Publication History

Received: 06 July 2011

Accepted after major revision: 08 February 2011

Publication Date:
22 November 2017 (online)

    Permissions and Reprints

Summary

The natural anticoagulant protein S contains a so-called thrombin-sensitive region (TSR), which is susceptible to proteolytic cleavage. We have previously shown that a platelet-associated protease is able to cleave protein S under physiological plasma conditions in vitro. The aim of the present study was to investigate the relation between platelet-associated protein S cleaving activity and in vivo protein S cleavage, and to evaluate the impact of in vivo protein S cleavage on its anticoagulant activity. Protein S cleavage in healthy subjects and in thrombocytopenic and thrombocythaemic patients was evaluated by immunological techniques. Concentration of cleaved and intact protein S was correlated to levels of activated protein C (APC)-dependent and APC-independent protein S anticoagulant activity. In plasma from healthy volunteers 25% of protein S is cleaved in the TSR. While in plasma there was a clear positive correlation between levels of intact protein S and both APC-dependent and APC-independent protein S anticoagulant activities, these correlations were absent for cleaved protein S. Protein S cleavage was significantly increased in patients with essential thrombocythaemia (ET) and significantly reduced in patients with chemotherapy-induced thrombocytopenia. In ET patients on cytoreductive therapy, both platelet count and protein S cleavage returned to normal values. Accordingly, platelet transfusion restored cleavage of protein S to normal values in patients with chemotherapy-induced thrombocytopenia. In conclusion, proteases from platelets seem to contribute to the presence of cleaved protein S in the circulation and may enhance the coagulation response in vivo by down regulating the anticoagulant activity of protein S.

Keywords

Coagulation - thrombin generation - protein S - cellular proteases - platelets
 

  • References

  • 1 Saller F, Brisset AC, Tchaikovski SN. et al. Generation and phenotypic analysis of protein S-deficient mice. Blood 2009; 114: 2307-2314.
    CrossrefPubMedGoogle Scholar
  • 2 Burstyn-Cohen T, Heeb MJ, Lemke G. Lack of Protein S in mice causes embryonic lethal coagulopathy and vascular dysgenesis. J Clin Invest 2009; 119: 2942-2953.
    CrossrefPubMedGoogle Scholar
  • 3 Bombeli T, Basic A, Fehr J. Prevalence of hereditary thrombophilia in patients with thrombosis in different venous systems. Am J Hematol 2002; 70: 126-132.
    CrossrefPubMedGoogle Scholar
  • 4 Mahmoodi BK, Brouwer JL, Veeger NJ. et al. Hereditary deficiency of protein C or protein S confers increased risk of arterial thromboembolic events at a young age: results from a large family cohort study. Circulation 2008; 118: 1659-1667.
    CrossrefPubMedGoogle Scholar
  • 5 Dahlbäck B, Villoutreix BO. Regulation of blood coagulation by the protein C anticoagulant pathway: novel insights into structure-function relationships and molecular recognition. Arterioscler Thromb Vasc Biol 2005; 25: 1311-1320.
    CrossrefPubMedGoogle Scholar
  • 6 Hackeng TM, Rosing J. Protein S as cofactor for TFPI. Arterioscler Thromb Vasc Biol 2009; 29: 2015-2020.
    CrossrefPubMedGoogle Scholar
  • 7 Heeb MJ, Prashun D, Griffin JH. et al. Plasma protein S contains zinc essential for efficient activated protein C-independent anticoagulant activity and binding to factor Xa, but not for efficient binding to tissue factor pathway inhibitor. FASEB J 2009; 23: 2244-2253.
    CrossrefPubMedGoogle Scholar
  • 8 Takeyama M, Nogami K, Saenko EL. et al. Protein S down-regulates factor Xase activity independent of activated protein C: specific binding of factor VIII(a) to protein S inhibits interactions with factor IXa. Br J Haematol 2008; 143: 409-420.
    CrossrefPubMedGoogle Scholar
  • 9 Long GL, Lu D, Xie RL, Kalafatis M. Human protein S cleavage and inactivation by coagulation factor Xa. J Biol Chem 1998; 273: 11521-11526.
    CrossrefPubMedGoogle Scholar
  • 10 Borgel D, Gaussem P, Garbay C. et al. Implication of protein S thrombin-sensitive region with membrane binding via conformational changes in the gamma-carbo-xyglutamic acid-rich domain. Biochem J 2001; 360: 499-506.
    CrossrefPubMedGoogle Scholar
  • 11 Heeb MJ, Griffin JH. Activated protein C-dependent and -independent anticoagulant activities of protein S have different structural requirements. Blood Cells Mol Dis 2002; 29: 190-199.
    CrossrefPubMedGoogle Scholar
  • 12 Brinkman HJ, Mertens K, van Mourik JA. Proteolytic cleavage of protein S during the hemostatic response. J Thromb Haemost 2005; 03: 2712-2720.
    CrossrefPubMedGoogle Scholar
  • 13 Morboeuf O, Borgel D, Gaussem P. et al. Characterization of cleaved plasma protein S with a monoclonal antibody-based assay. Thromb Haemost 2000; 84: 604-610.
    Article in Thieme ConnectPubMedGoogle Scholar
  • 14 Oates AM, Salem HH. The binding and regulation of protein S by neutrophils. Blood Coagul Fibrinolysis 1991; 02: 601-607.
    CrossrefPubMedGoogle Scholar
  • 15 Dahlbäck B. Purification of human vitamin K-dependent protein S and its limited proteolysis by thrombin. Biochem J 1983; 209: 837-846.
    CrossrefPubMedGoogle Scholar
  • 16 Eckle I, Seitz R, Egbring R. et al. Protein S degradation in vitro by neutrophil elas-tase. Scand J Clin Lab Invest 1993; 53: 281-288.
    CrossrefPubMedGoogle Scholar
  • 17 JMitchell CA, Salem HH. Cleavage of protein S by a platelet membrane protease. J Clin Invest 1987; 79: 374-379.
    CrossrefPubMedGoogle Scholar
  • 18 Marchetti M, Castoldi E, Spronk HM. et al. Thrombin generation and activated protein C resistance in patients with essential thrombocythemia and polycythe-mia vera. Blood 2008; 112: 4061-4068.
    CrossrefPubMedGoogle Scholar
  • 19 Meijer-Huizinga F, Mertens K, van Mourik JA. Isolation and characterization of single-chain protein S. Thromb Haemost 1994; 72: 408-414.
    Article in Thieme ConnectPubMedGoogle Scholar
  • 20 Griffin JH, Gruber A, Fernández JA. Reevaluation of total, free, and bound protein S and C4b-binding protein levels in plasma anticoagulated with citrate or hiru-din. Blood 1992; 79: 3203-3211.
    PubMedGoogle Scholar
  • 21 Hemker HC, Giesen P, Al Dieri R. et al. Calibrated automated thrombin generation measurement in clotting plasma. Pathophysiol Haemost Thromb 2003; 33: 4-15.
    CrossrefPubMedGoogle Scholar
  • 22 Seré KM, Rosing J, Hackeng TM. Inhibition of thrombin generation by protein S at low procoagulant stimuli: implications for maintenance of the hemostatic balance. Blood 2004; 104: 3624-3630.
    CrossrefPubMedGoogle Scholar
  • 23 Hackeng TM, Hessing M, van 't Veer C. et al. Protein S binding to human endothe-lial cells is required for expression of cofactor activity for activated protein C. J Biol Chem 1993; 268: 3993-4000.
    PubMedGoogle Scholar
  • 24 Hackeng TM, van 't Veer C, Meijers JC. et al. Human protein S inhibits prothrombinase complex activity on endothelial cells and platelets via direct interactions with factors Va and Xa. J Biol Chem 1994; 269: 21051-21058.
    PubMedGoogle Scholar
  • 25 Schwarz HP, Heeb MJ, Wencel-Drake JD, Griffin JH. Identification and quanti-tation of protein S in human platelets. Blood 1985; 66: 1452-1455.
    PubMedGoogle Scholar
  • 26 Borgel D, Reny JL, Fischelis D. et al. Cleaved protein S (PS), total PS, free PS, and activated protein C cofactor activity as risk factors for venous thromboembolism. Clin Chem 2003; 49: 575-580.
    CrossrefPubMedGoogle Scholar
  • 27 Damon AL, Scudder LE, Gnatenko D V. et al. Altered bioavailability of platelet-derived factor VIII during thrombocytosis reverses phenotypic efficacy in haemo-philic mice. Thromb Haemost 2008; 100: 1111-1122.
    Article in Thieme ConnectPubMedGoogle Scholar
  • 28 Castoldi E, Maurissen LF, Tormene D. et al. Similar hypercoagulable state and thrombosis risk in type I and type III protein S-deficient individuals from families with mixed type I/III protein S deficiency. Haematologica 2010; 95: 1563-1571.
    CrossrefPubMedGoogle Scholar
  • 29 Bertina RM, van Wijngaarden A, Reinalda-Poot J. et al. Determination of plasma protein S, the protein cofactor of activated protein C. Thromb Haemost 1985; 53: 268-272.
    Article in Thieme ConnectPubMedGoogle Scholar
  • 30 Malm K, Dahlbäck B, Arnljots B. Prevention of thrombosis following deep arterial injury in rats by bovine activated protein C requiring co-administration of bovine protein S. Thromb Haemost 2003; 90: 227-234.
    Article in Thieme ConnectPubMedGoogle Scholar
  • 31 Camire RM, Kalafatis M, Tracy PB. Proteolysis of factor V by cathepsin G and elas-tase indicates that cleavage at Arg1545 optimizes cofactor function by facilitating factor Xa binding. Biochemistry 1998; 37: 11896-11906.
    CrossrefPubMedGoogle Scholar
  • 32 Gale AJ, Rozenshteyn D. Cathepsin G, a leukocyte protease, activates coagulation factor VIII. Thromb Haemost 2008; 99: 44-51.
    Article in Thieme ConnectPubMedGoogle Scholar
  • 33 Plescia J, Altieri DC. Activation of Mac-1 (CD11b/CD18)-bound factor X by released cathepsin G defines an alternative pathway of leucocyte initiation of coagulation. Biochem J 1996; 319: 873-879.
    CrossrefPubMedGoogle Scholar
  • 34 Reinhardt C, von Brühl ML, Manukyan D. et al. Protein disulfide isomerase acts as an injury response signal that enhances fibrin generation via tissue factor activation. J Clin Invest 2008; 118: 1110-1122.
    PubMedGoogle Scholar
  • 35 Nogami K, Ogiwara K, Matsumoto T. et al. Mechanisms of human neutrophil elastase-catalysed inactivation of factor VIII(a). Thromb Haemost 2011; 105: 968-980.
    Article in Thieme ConnectPubMedGoogle Scholar