Thromb Haemost 2003; 89(01): 132-141
DOI: 10.1055/s-0037-1613552
Platelets and Blood Cells
Schattauer GmbH

Platelet phosphatidylserine exposure and procoagulant activity in clotting whole blood – different effects of collagen, TRAP and calcium ionophore A23187

Sofia Ramström
1   Department of Biomedicine and Surgery, Division of Clinical Chemistry, University Hospital, Linköping, Sweden
2   Forum Scientum Graduate School, Linköping University, Sweden
,
Mats Rånby
1   Department of Biomedicine and Surgery, Division of Clinical Chemistry, University Hospital, Linköping, Sweden
3   Global Hemostasis Institute MGR AB, Linköping, Sweden
,
Tomas L. Lindahl
4   Department of Clinical Chemistry, Laboratory Medicine, University Hospital, Linköping, Sweden
› Author Affiliations
Further Information

Publication History

Received 26 June 2002

Accepted after revision 11 November 2002

Publication Date:
09 December 2017 (online)

Summary

We have studied the effects of different platelet agonists on phosphatidylserine (PS) exposure and clotting times in blood without anticoagulants. Similar reductions in clotting time were obtained for collagen, TRAP-6 or calcium ionophore A23187 (50 μmol/L), in spite of huge differences in PS expression [6.7 ± 2.4%, 2.3 ± 0.5% and 99.9 ± 0.1%, respectively (mean ± SD, n = 5)]. Furthermore, the clotting times were much longer for samples with A23187 exposing the same amounts of PS as samples with collagen or TRAP-6. Annexin V reversed the clotting time reduction, but could not prevent coagulation. Addition of phospholipid vesicles containing 20% PS neither affected the clotting times nor induced clotting in recalcified, platelet-free plasma.

We conclude that platelet PS exposure is necessary, but not sufficient, for the coagulation amplification observed when platelets are stimulated via physiological receptors in a whole blood environment.

 
  • References

  • 1 Bevers EM, Comfurius P, Zwaal RF. Regulatory mechanisms in maintenance and modulation of transmembrane lipid asymmetry: pathophysiological implications. Lupus 1996; 5: 480-7.
  • 2 Stenflo J. Structure-function relationships of epidermal growth factor modules in vitamin K-dependent clotting factors. Blood 1991; 78: 1637-51.
  • 3 Sunnerhagen M, Drakenberg T, Forsen S, Stenflo J. Effect of Ca2+ on the structure of vitamin K-dependent coagulation factors. Haemostasis 1996; 26: 45-53.
  • 4 Blockmans D, Deckmyn H, Vermylen J. Platelet activation. Blood Rev 1995; 9: 143-56.
  • 5 Carr Jr. ME, Carr SL, Hantgan RR, Braaten J. Glycoprotein IIb/IIIa blockade inhibits platelet-mediated force development and reduces gel elastic modulus. Thromb Haemost 1995; 73: 499-505.
  • 6 Furman MI, Krueger LA, Frelinger AL, Barnard MR, Mascelli MA, Nakada MT, Michelson AD. GPIIb-IIIa antagonist-induced reduction in platelet surface factor V/Va binding and phosphatidylserine expression in whole blood. Thromb Haemost 2000; 84: 492-8.
  • 7 Ramström S, Rånby M, Lindahl T. The role of platelets in blood coagulation – effects of platelet agonists and of GPIIb/IIIa inhibitors studied by free oscillation rheometry. Thromb Res 2002; 105: 165-72.
  • 8 Bohlin L. Method of measuring rheological properties and rheometer for carrying out the method. In: PCT (Patent Cooperation Treaty) publication number WO94/08222 1994
  • 9 Bohlin L. Bearing device. In: PCT (Patent Cooperation Treaty) publication number WO98/54475. 1998
  • 10 Laudano AP, Cottrell BA, Doolittle RF. Synthetic peptides modeled on fibrin polymerization sites. Ann NY Acad Sci 1983; 408: 315-29.
  • 11 Kawasaki K, Hirase K, Miyano M, Tsuji T, Iwamoto M. Amino acids and peptides. XVI. Synthesis of N-terminal tetrapeptide analogs of fibrin α-chain and their inhibitory effects on fibrinogen/thrombin clotting. Chem Pharm Bull 1992; 40: 3253-60.
  • 12 Coughlin SR. How the protease thrombin talks to cells. Proc Natl Acad Sci USA 1999; 96: 11023-7.
  • 13 Kehrel B, Wierwille S, Clemetson KJ, Anders O, Steiner M, Knight CG, Farndale RW, Okuma M, Barnes MJ. Glycoprotein VI is a major collagen receptor for platelet activation: it recognizes the platelet-activating quaternary structure of collagen, whereas CD36, glyco-protein IIb/IIIa, and von Willebrand factor do not. Blood 1998; 91: 491-9.
  • 14 Lindahl TL, Festin R, Larsson A. Studies of fibrinogen binding to platelets by flow cytometry: an improved method for studies of platelet activation. Thromb Haemost 1992; 68: 221-5.
  • 15 Morrissey J. Morrissey Lab Protocol for Preparing Phospholipid Vesicles by Extrusion. At. http://tf7.org/Extrude.html Accessed 2001 09/06.
  • 16 Pedicord DL, Thomas BE, Mousa SA, Dicker IB. Glycoprotein IIb/IIIa receptor antagonists inhibit the development of platelet procoagulant activity. Thromb Res 1998; 90: 247-58.
  • 17 Pasquet JM, Dachary-Prigent J, Nurden AT. Microvesicle release is associated with extensive protein tyrosine dephosphorylation in platelets stimulated by A23187 or a mixture of thrombin and collagen. Biochem J 1998; 333: 591-9.
  • 18 Dachary-Prigent J, Pasquet J-M, Nurden A. Simultaneous detection of changes in cytoplasmic Ca2+, aminophospholipid exposure and microvesiculation in activated platelets. Platelets 1997; 8: 405-12.
  • 19 Dormann D, Clemetson KJ, Kehrel BE. The GPIb thrombin-binding site is essential for thrombin-induced platelet procoagulant activity. Blood 2000; 96: 2469-78.
  • 20 Storey RF, Sanderson HM, White AE, May JA, Cameron KE, Heptinstall S. The central role of the P(2T) receptor in amplification of human platelet activation, aggregation, secretion and procoagulant activity. Br J Haematol 2000; 110: 925-34.
  • 21 Beguin S, Kumar R, Keularts I, Seligsohn U, Coller BS, Hemker HC. Fibrin-dependent platelet procoagulant activity requires GPIb receptors and von Willebrand factor. Blood 1999; 93: 564-70.
  • 22 Alberio L, Safa O, Clemetson KJ, Esmon CT, Dale GL. Surface expression and functional characterization of alpha-granule factor V in human platelets: effects of ionophore A23187, thrombin, collagen, and convulxin. Blood 2000; 95: 1694-702.
  • 23 Siljander P, Farndale RW, Feijge MA, Comfurius P, Kos S, Bevers EM, Heemskerk JW. Platelet adhesion enhances the glycoprotein VI-dependent procoagulant response: Involvement of p38 MAP kinase and calpain. Arterioscler Thromb Vasc Biol 2001; 21: 618-27.
  • 24 Byzova TV, Plow EF. Networking in the hemostatic system. Integrin alphaiibbeta3 binds prothrombin and influences its activation. J Biol Chem 1997; 272: 27183-8.
  • 25 Lages B, Weiss HJ. Greater inhibition of platelet procoagulant activity by antibody-derived glycoprotein IIb-IIIa inhibitors than by peptide and peptidomimetic inhibitors. Br J Haematol 2001; 113: 65-71.
  • 26 Rand M, Lock J, van’t Veer C, Gaffney D, Mann K. Blood clotting in minimally altered whole blood. Blood 1996; 88: 3432-45.
  • 27 Dachary-Prigent J, Freyssinet JM, Pasquet JM, Carron JC, Nurden AT. Annexin V as a probe of aminophospholipid exposure and platelet membrane vesiculation: a flow cytometry study showing a role for free sulfhydryl groups. Blood 1993; 81: 2554-65.
  • 28 Sumner WT, Monroe DM, Hoffman M. Variability in platelet procoagulant activity in healthy volunteers. Thromb Res 1996; 81: 533-43.
  • 29 Niewiarowski S. Secreted platelet proteins. In: Haemostasis and Thrombosis. Bloom A, Forbes C, Thomas D, Tuddenham E. eds Churchill Livingstone 1994; 167-81.
  • 30 Holme PA, Brosstad F, Solum NO. Platelet-derived microvesicles and activated platelets express factor Xa activity. Blood Coagul Fibrinolysis 1995; 6: 302-10.
  • 31 Nesheim ME, Furmaniak-Kazmierczak E, Henin C, Cote G. On the existence of platelet receptors for factor V(a) and factor VIII(a). Thromb Haemost 1993; 70: 80-6.
  • 32 Bouchard BA, Catcher CS, Thrash BR, Adida C, Tracy PB. Effector cell protease receptor-1, a platelet activation-dependent membrane protein, regulates prothrombinase-catalyzed thrombin generation. J Biol Chem 1997; 272: 9244-51.
  • 33 London F, Ahmad SS, Walsh PN. Annexin V inhibition of factor IXa-catalyzed factor X activation on human platelets and on negatively-charged phospholipid vesicles. Biochemistry 1996; 35: 16886-97.
  • 34 Solum NO. Procoagulant expression in platelets and defects leading to clinical disorders. Arterioscler Thromb Vasc Biol 1999; 19: 2841-6.
  • 35 Stormorken H, Holmsen H, Sund R, Sakariassen KS, Hovig T, Jellum E, Solum O. Studies on the haemostatic defect in a complicated syndrome. An inverse Scott syndrome platelet membrane abnormality?. Thromb Haemost 1995; 74: 1244-51.