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

Merel van Wijnen; Cornelis  van ’t Veer; Joost C.M. Meijers; Rogier M. Bertina; Bonno N. Bouma

doi:10.1055/s-0037-1615391

Thrombosis and Haemostasis, Table of Contents

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

Authors

Merel van Wijnen^*

¹Department of Haematology, University Hospital Utrecht

²Institute of Biomembranes, Utrecht University, The Netherlands
Cornelis van ’t Veer

¹Department of Haematology, University Hospital Utrecht

⁴Present address: Dr. C. van ’t Veer, Department of Surgery, Maastricht University, The Netherlands
Joost C.M. Meijers

¹Department of Haematology, University Hospital Utrecht

²Institute of Biomembranes, Utrecht University, The Netherlands
Rogier M. Bertina

³Thrombosis and Haemostasis Research Center, Department of Haematology, Leiden University Medical Center, The Netherlands
Bonno N. Bouma

¹Department of Haematology, University Hospital Utrecht

²Institute of Biomembranes, Utrecht University, The Netherlands

Abstract

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.

Full Text

References

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.
2 Bertina RM. Hereditary protein S deficiency. Haemostasis 1985; 15: 241-6.
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.
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.
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.
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.
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.
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.
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 V^R506Q mutation. Thromb Haemost 1996; 76: 208-14.
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.
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.
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.
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.
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.
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.
16 van’t Veer C, Golden NJ, Mann KG. Regulation of prothrombinase activity by protein S. Thromb Haemost 1997; supplement: 186-7 (abstract).
17 Walker FJ. Regulation of vitamin K-dependent protein S. Inactivation by thrombin. J Biol Chem 1984; 259: 10335-9.
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.
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.
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.
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.
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.
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.
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.
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.
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.
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.
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.
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.
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.
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.
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.
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.
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.
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.