Download PDF
Thromb Haemost 1998; 80(06): 930-935
DOI: 10.1055/s-0037-1615391
Letters to the Editor
Schattauer GmbH

A Plasma Coagulation Assay for an Activated Protein C-independent Anticoagulant Activity of Protein S

Merel van Wijnen   *
1   Department of Haematology, University Hospital Utrecht
2   Institute of Biomembranes, Utrecht University, The Netherlands
,
Cornelis van ’t Veer
1   Department of Haematology, University Hospital Utrecht
4   Present address: Dr. C. van ’t Veer, Department of Surgery, Maastricht University, The Netherlands
,
Joost C.M. Meijers
1   Department of Haematology, University Hospital Utrecht
2   Institute of Biomembranes, Utrecht University, The Netherlands
,
Rogier M. Bertina
3   Thrombosis and Haemostasis Research Center, Department of Haematology, Leiden University Medical Center, The Netherlands
,
Bonno N. Bouma
1   Department of Haematology, University Hospital Utrecht
2   Institute of Biomembranes, Utrecht University, The Netherlands
› Author Affiliations
Further Information

Publication History

Received 29 June 1998

Accepted after revision 01 September 1998

Publication Date:
07 December 2017 (online)

Also available at
    Permissions and Reprints

Summary

To study the physiological importance of the activated protein C (APC)-independent anticoagulant activity of protein S, we developed an assay specific for this activity. The ability of protein S to prolong the clotting time in an APC-independent way was expressed as the ratio of the clotting time in a plasma sample divided by the clotting time in the presence of a polyclonal antibody against human protein S (both after 1:1 dilution in protein S-C4BP deficient plasma). The mean protein S-dependent anticoagulant ratio (PSdAR) was 1.53 ± 0.18 in 34 healthy controls, and was significantly lower in 16 heterozygous protein S deficient patients (PSdAR = 1.15 ± 0.09). In plasmas from patients under oral anticoagulant therapy the mean PSdAR was within the range of the control population (1.50 ± 0.18). The mean total protein S antigen level in these plasmas was 58%, suggesting a higher specific APC-independent anticoagulant activity of protein S in these patients than in normals.

This functional protein S assay can be used for the laboratory diagnosis of protein S deficiency, and to study the mechanism of the APC-independent anticoagulant activity in plasma.

* M. van Wijnen is supported by a grant from the Netherlands Heart Foundation (93.108). J.C.M. Meijers is an Established Investigator of the Netherlands Heart Foundation.


 

  • References

  • 1 Schwarz HP, Fischer M, Hopmeier P, Batard MA, Griffin JH. Plasma protein S deficiency in familial thrombotic disease. Blood 1984; 64: 1297-300.
    PubMedSearch in Google Scholar
  • 2 Bertina RM. Hereditary protein S deficiency. Haemostasis 1985; 15: 241-6.
    PubMedSearch in Google Scholar
  • 3 Comp PC, Esmon CT. Recurrent venous thromboembolism in patients with a partial deficiency of protein S. N Engl J Med 1984; 311: 1525-8.
    CrossrefPubMedSearch in Google Scholar
  • 4 Comp PC, Nixon RR, Cooper MR, Esmon CT. Familial protein S deficiency is associated with recur-rent thrombosis. J Clin Invest 1984; 74: 2082-8.
    CrossrefPubMedSearch in Google Scholar
  • 5 Rosing J, Hoekema L, Nicolaes GAF, Christella M, Thomassen LGD, Hemker HC, Varadi K, Schwarz HP, Tans G. Effects of protein S and factor Xa on peptide bond cleavages during inactivation of factor Va and factor VaR506Q by activated protein C. J Biol Chem 1995; 270: 27852-8.
    CrossrefPubMedSearch in Google Scholar
  • 6 Regan LM, Lamphear BJ, Huggins CF, Walker FJ, Fay PJ. Factor IXa protects factor VIIIa from activated protein C. Factor IXa inhibits activated protein C-catalyzed cleavage of factor VIIIa at Arg562 . J Biol Chem 1994; 269: 9445-52.
    PubMedSearch in Google Scholar
  • 7 Shen L, Dahlbäck B. Factor V and protein S as synergistic cofactors to activated protein C in degrada-tion of factor VIIIa. J Biol Chem 1994; 269: 18735-8.
    PubMedSearch in Google Scholar
  • 8 Lu D, Kalafatis M, Mann KG, Long GL. Comparison of activated protein C/protein S-mediated inactivation of human factor VIII and factor V. Blood 1996; 87: 4708-17.
    PubMedSearch in Google Scholar
  • 9 Varadi K, Rosing J, Tans G, Pabinger I, Keil B, Schwarz HP. Factor V enhances the cofactor function of protein S in the APC-mediated inactivation of factor VIII: influence of the factor VR506Q mutation. Thromb Haemost 1996; 76: 208-14.
    Thieme ConnectPubMedSearch in Google Scholar
  • 10 Heeb MJ, Rosing J, Bakker HM, Fernandez JA, Tans G, Griffin JH. Protein S binds to and inhibits factor Xa. Proc Natl Acad Sci U S A 1994; 91: 2728-32.
    CrossrefPubMedSearch in Google Scholar
  • 11 Heeb MJ, Mesters RM, Tans G, Rosing J, Griffin JH. Binding of protein S to factor Va associated with inhibition of prothrombinase that is independent of activated protein C. J Biol Chem 1993; 268: 2872-7.
    PubMedSearch in Google Scholar
  • 12 Koppelman SJ, Hackeng TM, Sixma JJ, Bouma BN. Inhibition of the intrinsic factor X activating complex by protein S: evidence for a specific binding of protein S to factor VIII. Blood 1995; 86: 1062-71.
    CrossrefPubMedSearch in Google Scholar
  • 13 Hackeng TM, van’t Veer C, Meijers JC, Bouma BN. 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: 21051-8.
    PubMedSearch in Google Scholar
  • 14 van’t Veer C, Hackeng TM, Biesbroeck D, Sixma JJ, Bouma BN. Increased prothrombin activation in protein S-deficient plasma under flow conditions on endothelial cell matrix: an independent anticoagulant function of protein S in plasma. Blood 1995; 85: 1815-21.
    PubMedSearch in Google Scholar
  • 15 van Wijnen M, Stam JG, van’t Veer C, Meijers JC, Reitsma PH, Bertina RM, Bouma BN. The interaction of protein S with the phospholipid surface is essential for the activated protein C-independent activity of protein S. Thromb Haemost 1996; 76: 397-403.
    Thieme ConnectPubMedSearch in Google Scholar
  • 16 van’t Veer C, Golden NJ, Mann KG. Regulation of prothrombinase activity by protein S. Thromb Haemost 1997; supplement: 186-7 (abstract).
    PubMedSearch in Google Scholar
  • 17 Walker FJ. Regulation of vitamin K-dependent protein S. Inactivation by thrombin. J Biol Chem 1984; 259: 10335-9.
    PubMedSearch in Google Scholar
  • 18 Bertina RM, van Wijngaarden A, Reinalda Poot J, Poort SR, Bom VJ. Determination of plasma protein S--the protein cofactor of activated protein C. Thromb Haemost 1985; 53: 268-72.
    Thieme ConnectPubMedSearch in Google Scholar
  • 19 Dahlbäck B. Inhibition of protein Ca cofactor function of human and bovine protein S by C4b-binding protein. J Biol Chem 1986; 261: 12022-7.
    PubMedSearch in Google Scholar
  • 20 Preda L, Simioni P, Legnani C, Palareti G, Poggio M, Rossi E, Girolami A. A new global test for the evaluation of the protein C-protein S system. Blood Coagul Fibrinolysis 1996; 7: 465-9.
    CrossrefPubMedSearch in Google Scholar
  • 21 Lefkowitz JB, Clarke SH, Barbour LA. Comparison of protein S functional and antigenic assays in normal pregnancy. Am J Obstet Gynecol 1996; 175: 657-60.
    CrossrefPubMedSearch in Google Scholar
  • 22 Boyer Neumann C, Bertina RM, Tripodi A, D’Angelo A, Wolf M, Vigano D, Mannucci PM, Meyer D, Larrieu MJ. Comparison of functional assays for protein S: European collaborative study of patients with congenital and acquired deficiency. Thromb Haemost 1993; 70: 946-50.
    Thieme ConnectPubMedSearch in Google Scholar
  • 23 Malm J, Laurell M, Dahlbäck B. Changes in the plasma levels of vitamin K-dependent proteins C and S and of C4b-binding protein during pregnancy and oral contraception. Br J Haematol 1988; 68: 437-43.
    CrossrefPubMedSearch in Google Scholar
  • 24 Deutz-Terlouw PP, Ballering L, van Wijngaarden A, Bertina RM. Two ELISA’s for measurement of protein S, and their use in the laboratory diagnosis of protein S deficiency. Clin Chim Acta 1989; 186: 321-34.
    PubMedSearch in Google Scholar
  • 25 Hessing M, Kanters D, Hackeng TM, Bouma BN. Identification of different forms of human C4b-binding protein lacking beta-chain and protein S binding ability. Thromb Haemost 1990; 64: 245-50.
    Thieme ConnectPubMedSearch in Google Scholar
  • 26 Perkins SJ, Chung LP, Reid KB. Unusual ultrastructure of complement-component-C4b-binding protein of human complement by synchrotron X-ray scattering and hydrodynamic analysis. Biochem J 1986; 233: 799-807.
    CrossrefPubMedSearch in Google Scholar
  • 27 DiScipio RG, Davie EW. Characterization of protein S, a gamma-carboxyglutamic acid containing protein from bovine and human plasma. Biochemistry 1979; 18: 899-904.
    CrossrefPubMedSearch in Google Scholar
  • 28 Hessing M, Kanters D, Heijnen HF, Hackeng TM, Sixma JJ, Bouma BN. Structure-function studies on human C4b-binding protein using monoclonal antibodies. Eur J Immunol 1991; 21: 2077-85.
    CrossrefPubMedSearch in Google Scholar
  • 29 Reitsma PH, Ploos van Amstel HK, Bertina RM. Three novel mutations in five unrelated subjects with hereditary protein S deficiency type I. J Clin Invest 1994; 93: 486-92.
    CrossrefPubMedSearch in Google Scholar
  • 30 Chang GT, Ploos van Amstel HK, Hessing M, Reitsma PH, Bertina RM, Bouma BN. Expression and characterization of recombinant human protein S in heterologous cells. Studies of the interaction of amino acid residues leu-608 to glu-612 with human C4b-binding protein. Thromb Haemost 1992; 67: 526-32.
    Thieme ConnectPubMedSearch in Google Scholar
  • 31 D’Angelo A, Vigano D, Esmon CT, Comp PC. Acquired deficiencies of protein S. Protein S activity during oral anticoagulation, in liver disease, and in disseminated intravascular coagulation. J Clin Invest 1988; 81: 1445-54.
    CrossrefPubMedSearch in Google Scholar
  • 32 Zwaginga JJ, Sixma JJ, de Groot PG. Activation of endothelial cells induces platelet thrombus formation on their matrix. Studies of new in vitro thrombosis model with low molecular weight heparin as anticoagulant. Artherioscl 1990; 10: 49-61.
    PubMedSearch in Google Scholar
  • 33 Marciniak E. Factor Xa inactivation by antithrombin III: Evidence for biological stabilization of factor Xa by factor V phospholipid complex. Br J Haematol 1973; 24: 391.
    CrossrefPubMedSearch in Google Scholar
  • 34 Walker FJ, Esmon CT. The effects of phospholipid and factor Va on the inhibition of factor Xa by antithrombin III. Biochem Biophys Res Commun 1979; 90: 641-7.
    CrossrefPubMedSearch in Google Scholar
  • 35 Hackeng TM, Hessing M, van ’t Veer C, Meijer Huizinga F, Meijers JC, de Groot PG, van Mourik JA, Bouma BN. Protein S binding to human endothelial cells is required for expression of cofactor activity for activated protein C. J Biol Chem 1993; 268: 3993-4000.
    PubMedSearch in Google Scholar