Thromb Haemost 2018; 118(07): 1194-1202
DOI: 10.1055/s-0038-1656549
Coagulation and Fibrinolysis
Georg Thieme Verlag KG Stuttgart · New York

Factor V Has Anticoagulant Activity in Plasma in the Presence of TFPIα: Difference between FV1 and FV2

Peter van Doorn
1   Department of Biochemistry, Cardiovascular Research Institute Maastricht, Maastricht University, The Netherlands
,
Jan Rosing
1   Department of Biochemistry, Cardiovascular Research Institute Maastricht, Maastricht University, The Netherlands
,
Connie Duckers
1   Department of Biochemistry, Cardiovascular Research Institute Maastricht, Maastricht University, The Netherlands
,
Tilman M. Hackeng
1   Department of Biochemistry, Cardiovascular Research Institute Maastricht, Maastricht University, The Netherlands
,
Paolo Simioni
2   Thrombotic and Hemorrhagic Diseases Unit, Department of Medicine, University of Padua Medical School, Padua, Italy
,
Elisabetta Castoldi
1   Department of Biochemistry, Cardiovascular Research Institute Maastricht, Maastricht University, The Netherlands
› Author Affiliations
Funding This work was supported by grant 2014–1 from the Dutch Thrombosis Foundation. Prof. Jan Rosing is Van de Laar Professor of Coagulation Enzymology.
Further Information

Publication History

09 January 2018

19 April 2018

Publication Date:
04 June 2018 (online)

Abstract

Background Activated factor V (FVa) is a potent procoagulant cofactor in the prothrombinase complex, whereas its precursor factor V (FV) stimulates the inhibition of factor Xa (FXa) by tissue factor pathway inhibitor-α (TFPIα), presumably by promoting TFPIα binding to phospholipids. Plasma FV comprises two glycosylation isoforms (FV1 and FV2) with low and high phospholipid-binding affinity, respectively. The FV1/FV2 ratio is increased in carriers of the FV R2 haplotype.

Objective This article demonstrates the TFPIα-cofactor function of FV in plasma and compares FV1 and FV2.

Materials and Methods Thrombin generation at low TF concentration was measured in FV-depleted plasma reconstituted with 0 to 100% FV, FV1 or FV2, and in 122 individuals genotyped for the R2 haplotype. The TFPIα-cofactor activities of FV1 and FV2 were also investigated in a model system of TFPIα-mediated FXa inhibition.

Results In the FV titration, thrombin generation first increased (up to 5% FV) and then progressively decreased at higher FV concentrations. This anticoagulant effect of FV, which was also observed with FV2 but not with FV1, was largely abolished by anti-TFPIα antibodies, suggesting that it reflects TFPIα-cofactor activity of FV. In the model system of TFPIα-mediated FXa inhibition, FV2 was a more potent TFPIα-cofactor than FV1, in line with their respective phospholipid affinities. Accordingly, FV R2 carriers had higher thrombin generation than non-carriers, even after correction for demographics and plasma levels of coagulation factors and inhibitors.

Conclusion FV (and particularly its FV2 isoform) contributes to the TFPIα-dependent down-regulation of thrombin generation in plasma triggered with low TF.

Supplementary Material

 
  • References

  • 1 Dahlbäck B. Novel insights into the regulation of coagulation by factor V isoforms, tissue factor pathway inhibitorα, and protein S. J Thromb Haemost 2017; 15 (07) 1241-1250
  • 2 Monkovic DD, Tracy PB. Activation of human factor V by factor Xa and thrombin. Biochemistry 1990; 29 (05) 1118-1128
  • 3 Camire RM, Bos MH. The molecular basis of factor V and VIII procofactor activation. J Thromb Haemost 2009; 7 (12) 1951-1961
  • 4 Rosing J, Tans G, Govers-Riemslag JW, Zwaal RF, Hemker HC. The role of phospholipids and factor Va in the prothrombinase complex. J Biol Chem 1980; 255 (01) 274-283
  • 5 Dahlbäck B, Villoutreix BO. The anticoagulant protein C pathway. FEBS Lett 2005; 579 (15) 3310-3316
  • 6 Camire RM. Rethinking events in the haemostatic process: role of factor V and TFPI. Haemophilia 2016; 22 (Suppl. 05) 3-8
  • 7 Shen L, Dahlbäck B. Factor V and protein S as synergistic cofactors to activated protein C in degradation of factor VIIIa. J Biol Chem 1994; 269 (29) 18735-18738
  • 8 Thorelli E, Kaufman RJ, Dahlbäck B. Cleavage requirements for activation of factor V by factor Xa. Eur J Biochem 1997; 247 (01) 12-20
  • 9 Vincent LM, Tran S, Livaja R, Bensend TA, Milewicz DM, Dahlbäck B. Coagulation factor V(A2440G) causes east Texas bleeding disorder via TFPIα. J Clin Invest 2013; 123 (09) 3777-3787
  • 10 Duckers C, Simioni P, Spiezia L. , et al. Low plasma levels of tissue factor pathway inhibitor in patients with congenital factor V deficiency. Blood 2008; 112 (09) 3615-3623
  • 11 Mast AE, Broze Jr GJ. Physiological concentrations of tissue factor pathway inhibitor do not inhibit prothrombinase. Blood 1996; 87 (05) 1845-1850
  • 12 Peraramelli S, Thomassen S, Heinzmann A. , et al. Role of exosite binding modulators in the inhibition of Fxa by TFPI. Thromb Haemost 2016; 115 (03) 580-590
  • 13 Santamaria S, Reglińska-Matveyev N, Gierula M. , et al. Factor V has an anticoagulant cofactor activity that targets the early phase of coagulation. J Biol Chem 2017; 292 (22) 9335-9344
  • 14 Ndonwi M, Girard TJ, Broze Jr GJ. The C-terminus of tissue factor pathway inhibitor α is required for its interaction with factors V and Va. J Thromb Haemost 2012; 10 (09) 1944-1946
  • 15 Wood JP, Bunce MW, Maroney SA, Tracy PB, Camire RM, Mast AE. Tissue factor pathway inhibitor-alpha inhibits prothrombinase during the initiation of blood coagulation. Proc Natl Acad Sci U S A 2013; 110 (44) 17838-17843
  • 16 Mast AE. Tissue factor pathway inhibitor: multiple anticoagulant activities for a single protein. Arterioscler Thromb Vasc Biol 2016; 36 (01) 9-14
  • 17 Ndonwi M, Tuley EA, Broze Jr GJ. The Kunitz-3 domain of TFPI-alpha is required for protein S-dependent enhancement of factor Xa inhibition. Blood 2010; 116 (08) 1344-1351
  • 18 Hackeng TM, Seré KM, Tans G, Rosing J. Protein S stimulates inhibition of the tissue factor pathway by tissue factor pathway inhibitor. Proc Natl Acad Sci U S A 2006; 103 (09) 3106-3111
  • 19 Ndonwi M, Broze Jr G. Protein S enhances the tissue factor pathway inhibitor inhibition of factor Xa but not its inhibition of factor VIIa-tissue factor. J Thromb Haemost 2008; 6 (06) 1044-1046
  • 20 Ahnström J, Andersson HM, Hockey V. , et al. Identification of functionally important residues in TFPI Kunitz domain 3 required for the enhancement of its activity by protein S. Blood 2012; 120 (25) 5059-5062
  • 21 Maurissen LF, Castoldi E, Simioni P, Rosing J, Hackeng TM. Thrombin generation-based assays to measure the activity of the TFPI-protein S pathway in plasma from normal and protein S-deficient individuals. J Thromb Haemost 2010; 8 (04) 750-758
  • 22 Rosing J, Bakker HM, Thomassen MC, Hemker HC, Tans G. Characterization of two forms of human factor Va with different cofactor activities. J Biol Chem 1993; 268 (28) 21130-21136
  • 23 Nicolaes GA, Villoutreix BO, Dahlbäck B. Partial glycosylation of Asn2181 in human factor V as a cause of molecular and functional heterogeneity. Modulation of glycosylation efficiency by mutagenesis of the consensus sequence for N-linked glycosylation. Biochemistry 1999; 38 (41) 13584-13591
  • 24 Hoekema L, Nicolaes GA, Hemker HC, Tans G, Rosing J. Human factor Va1 and factor Va2: properties in the procoagulant and anticoagulant pathways. Biochemistry 1997; 36 (11) 3331-3335
  • 25 Váradi 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 (02) 208-214
  • 26 Lunghi B, Iacoviello L, Gemmati D. , et al. Detection of new polymorphic markers in the factor V gene: association with factor V levels in plasma. Thromb Haemost 1996; 75 (01) 45-48
  • 27 Bernardi F, Faioni EM, Castoldi E. , et al. A factor V genetic component differing from factor V R506Q contributes to the activated protein C resistance phenotype. Blood 1997; 90 (04) 1552-1557
  • 28 Castoldi E, Rosing J, Girelli D. , et al. Mutations in the R2 FV gene affect the ratio between the two FV isoforms in plasma. Thromb Haemost 2000; 83 (03) 362-365
  • 29 Hoekema L, Castoldi E, Tans G. , et al. Functional properties of factor V and factor Va encoded by the R2-gene. Thromb Haemost 2001; 85 (01) 75-81
  • 30 Alhenc-Gelas M, Nicaud V, Gandrille S. , et al. The factor V gene A4070G mutation and the risk of venous thrombosis. Thromb Haemost 1999; 81 (02) 193-197
  • 31 de Visser MC, Guasch JF, Kamphuisen PW, Vos HL, Rosendaal FR, Bertina RM. The HR2 haplotype of factor V: effects on factor V levels, normalized activated protein C sensitivity ratios and the risk of venous thrombosis. Thromb Haemost 2000; 83 (04) 577-582
  • 32 Nicolaes GAF, Tans G, Thomassen MCLGD. , et al. Peptide bond cleavages and loss of functional activity during inactivation of factor Va and factor VaR506Q by activated protein C. J Biol Chem 1995; 270 (36) 21158-21166
  • 33 Hemker HC, Giesen P, AlDieri R. , et al. The calibrated automated thrombogram (CAT): a universal routine test for hyper- and hypocoagulability. Pathophysiol Haemost Thromb 2002; 32 (5-6): 249-253
  • 34 van Doorn P, Rosing J, Wielders SJ, Hackeng TM, Castoldi E. The C-terminus of tissue factor pathway inhibitor-α inhibits factor V activation by protecting the Arg1545 cleavage site. J Thromb Haemost 2017; 15 (01) 140-149
  • 35 Hackeng TM, Suijlen DPL, Hemker HC. , et al. Thermostable inhibitors of activation of the blood clotting system through contact with foreign surfaces. Google Patents; 2013
  • 36 Segers O, Simioni P, Tormene D. , et al. Genetic modulation of the FV(Leiden)/normal FV ratio and risk of venous thrombosis in factor V Leiden heterozygotes. J Thromb Haemost 2012; 10 (01) 73-80
  • 37 Segers O, Simioni P, Tormene D, Castoldi E. Influence of single nucleotide polymorphisms on thrombin generation in factor V Leiden heterozygotes. Thromb Haemost 2014; 111 (03) 438-446
  • 38 Hackeng TM, Rosing J. Protein S as cofactor for TFPI. Arterioscler Thromb Vasc Biol 2009; 29 (12) 2015-2020
  • 39 Dielis AW, Castoldi E, Spronk HM. , et al. Coagulation factors and the protein C system as determinants of thrombin generation in a normal population. J Thromb Haemost 2008; 6 (01) 125-131
  • 40 Castoldi E, Brugge JM, Nicolaes GA, Girelli D, Tans G, Rosing J. Impaired APC cofactor activity of factor V plays a major role in the APC resistance associated with the factor V Leiden (R506Q) and R2 (H1299R) mutations. Blood 2004; 103 (11) 4173-4179
  • 41 Al Dieri R, Bloemen S, Kelchtermans H, Wagenvoord R, Hemker HC. A new regulatory function of activated factor V: inhibition of the activation by tissue factor/factor VII(a) of factor X. J Thromb Haemost 2013; 11 (03) 503-511
  • 42 Suehisa E, Kawasaki T, Toku M, Hidaka Y. Low level of factor V is associated with development of deep-vein thrombosis in Japanese patients. Thromb Res 2010; 125 (02) 128-133