The Role of Factor VIII in the Activation of Human Blood Coagulation Factor X by Activated Factor IX

Summary

The role of factor VIII in the activation of human factor X by factor IXa, Ca²⁺ and phospholipid has been investigated. Factor VIII stimulated the factor Xa formation after activation by factor Xa or thrombin; the activity of thrombin-activated factor VIII was about 4-fold that of factor Xa-activated factor VIII. The isolated procoagulant moiety of the factor VIII complex behaved identically to the complete complex, whereas the von Willebrand factor moiety did not participate in the factor Xa formation. Thrombin-activated factor VIII complex (factor Villa) was used to study the effect of factor Villa in kinetic experiments. The results revealed a complex kinetic behaviour, including substrate inhibition and non-linearity of the reaction rate with the enzyme concentration. Using previously obtained insight into the kinetics of factor X activation in the absence of factor VIII, the results were found to support the hypothesis that factor Villa participates in the factor Xa formation in a complex with phospholipid-bound factor IXa; the formation of the factor VUIa-factor IXa complex then increases the catalytic efficiency of the factor IXa by 500-fold.

Keywords

Factor VIII - Factor IX - Factor X - Intrinsic pathway Summary

• References

• 1 Jackson CM, Nemerson Y. Blood coagulation. Annu Rev Biochem 1980; 49: 765-811
• 2 Hoyer LW. The factor VIII complex: structure and function. Blood 1981; 58: 1-13
• 3 Zimmerman TS, Meyer D. Structure and function of factor VUI/von Willebrand factor. In: Haemostasis and Thrombosis Bloom AL, Thomas DP. (eds.). pp. 111-123 Churchill Livingstone; Edinburgh: 1981
• 4 Tuddenham E GD, Trabold NC, Collins JA, Hoyer LW. The properties of factor VIII coagulant activity prepared by immuno-adsorbent chromatography. J Lab Clin Med 1979; 93: 40-53
• 5 Harris RB, Johnson AJ, Hodgins LT. Partial purification of biologically active, low molecular weight, human antihemophilic factor free of von Willebrand factor. Biochim Biophys Acta 1981; 668: 471-480
• 6 Fay PJ, Chavin SI, Schroeder D, Young FE, Marder VJ. Purification and characterization of a highly purified human factor VIII consisting of a single type of polypeptide chain. Proc Natl Acad Sci U.S.A 1982; 79: 7200-7204
• 7 Fulcher CA, Zimmerman TS. Characterization of the human factor VIII procoagulant protein with a heterologous precipitating antibody. Proc Natl Acad Sci U.S.A 1982; 79: 1648-1652
• 8 Vehar GA, Davie EW. Preparation and properties of bovine factor VIII (antihemophilic factor). Biochemistry 1980; 19: 401-410
• 9 Fass DN, Knutson GJ, Katzmann JA. Monoclonal antibodies to porcine factor VIII coagulant and their use in the isolation of active coagulant protein. Blood 1982; 59: 594-600
• 10 Knutson GJ, Fass DN. Porcine factor VIII:C prepared by affinity interaction with von Willebrand factor and heterologous antibodies: sodium dodecyl sulfate polyacrylamide gel analysis. Blood 1982; 59: 615-624
• 11 Wood WI, Capon DJ, Simonsen CC, Eaton DL, Gitschier J, Keyt B, Seeburg PH, Smith DH, Hollingshead P, Wion KL, Delwart E, Tuddenham E GD, Vehar GA, Lawn RM. Expression of active human factor VIII from recombinant DNA clones. Nature 1984; 312: 330-337
• 12 Vehar GA, Keyt B, Eaton D, Rodriguez H, O’Brien DP, Rotblatt F, Oppermann H, Keck R, Wood WI, Harkins RN, Tuddenham E GD, Lawn RM, Capon DJ. Structure of human factor VIII. Nature 1984; 312: 337-342
• 13 Toole JJ, Knopf JL, Wozney JM, Sultzman LA, Buecker JL, Pittman DD, Kaufman RJ, Brown E, Shoemaker C, Orr EC, Amphlett GW, Foster WB, Coe ML, Knutson GJ, Fass DN, Hewick RM. Molecular cloning of a cDNA encoding human antihaemophilic factor. Nature 1984; 312: 342-347
• 14 Hemker HC, Kahn MJ P. Reaction sequence of blood coagulation. Nature (London) 1967; 215: 1201-1202
• 15 Österud B, Rapaport SI. Synthesis of the intrinsic factor X activator. Inhibition of the function of formed activator by antibodies to factor VIII and to factor IX. Biochemistry 1970; 9: 1854-1861
• 16 Chuang TF, Sargeant RB, Hougie C. The intrinsic activation of factor X in blood coagulation. Biochim Biophys Acta 1972; 273: 287-291
• 17 Rapaport SI, Schiffman S, Patch MJ, Ames SB. The importance of activation of antihaemophilic globulin and proaccelerin by traces of thrombin in the generation of intrinsic prothrombinase activity. Blood 1963; 21: 221-236
• 18 Macfarlane RG, Biggs R, Ash BJ, Denson K WE. The interaction of factors VIII and IX. Br J Haematol 1964; 10: 530-541
• 19 Davie EW, Fujikawa K, Legaz ME, Kato H. Role of proteases in blood coagulation. In: Proteases and Biological Control Reich H, Rifkin DB, Shaw E. (eds.). Vol. 2 65-77 Cold Spring Harbor Laboratory; Cold Spring Harbor: 1975
• 20 Mertens K, Bertina RM. Activation of human coagulation factor VIII by activated factor X, the common product of the intrinsic and the extrinsic pathway of blood coagulation. Thromb Haemostas 1982; 47: 96-100
• 21 van Dieijen G, Tans G, Rosing J, Hemker HC. The role of phospholipid and factor Villa in the activation of bovine factor X. J Biol Chem 1981; 256: 3433-3442
• 22 Hultin MB. Role of human factor VIII in factor X activation. J Clin Invest 1982; 69: 950-958
• 23 Link RP, Castellino FJ. Kinetic comparison of bovine blood coagulation factors IXaα and IXaβ toward bovine factor X. Biochemistry 1983; 22: 4033-4041
• 24 Neal GG, Esnouf MP. Kinetic studies of the activation of factor X by factors IXa and VIII:C in the absence of thrombin. Br J Haematol 1984; 57: 123-131
• 25 Griffith MJ, Reisner HM, Lundblad RL, Roberts HR. Measurement of human factor IXa activity in an isolated factor X activation system. Thromb Res 1982; 27: 289-301
• 26 Mertens K, Cupers R, van Wijngaarden A, Bertina RM. Binding of human blood-coagulation factors IXa and X to phospholipid membranes. Biochem J 1984; 223: 599-605
• 27 Mertens K, Bertina RM. The contribution of Ca²⁺ and phospholipids to the activation of human blood-coagulation factor X by activated factor IX. Biochem J 1984; 223: 607-615
• 28 Hendrix H, Lindhout T, Mertens K, Engels W, Hemker HC. Activation of human prothrombin by stoichiometric levels of staphy-locoagulase. J Biol Chem 1983; 258: 3637-3644
• 29 Lundblad RL, Uhteg LC, Vogel CN, Kingdon HS, Mann KG. Preparation and partial characterization of two forms of bovine thrombin. Biochem Biophys Res Commun 1975; 66: 482-489
• 30 Muller HP, van Tilburg NH, Bertina RM, Veltkamp JJ. Immunologic studies on the relationship between FVIII related antigen and FVIII procoagulant activity. Thromb Res 1980; 20: 85-96
• 31 Veltkamp JJ, Drion EF, Loeliger EA. Detection of the carrier state in hereditary coagulation disorders. I. Thrombos Diathes Haemorrh 1968; 19: 279-303
• 32 Muller HP, van Tilburg NH, Bertina RM, Terwiel JPh, Veltkamp JJ. Immunoradiometric assay of procoagulant factor VIII antigen (VIIICAg). Clin Chim Acta 1980; 107: 11-19
• 33 Weiss HJ, Hoyer LW, Rickless FR, Varma A, Rogers J. Quantitative assay of a plasma factor deficient in von Willebrand’s disease that is necessary for platelet aggregation. Relationship to factor VIII procoagulant activity and antigen content. J Clin Invest 1973; 52: 2708-2716
• 34 Veltkamp JJ, van Tilburg NH. Detection of heterozygotes for recessive von Willebrand’s disease by the assay of antihemophilic-factor-like antigen. N Engl J Med 1973; 289: 882-885
• 35 Bradford MM. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem 1976; 72: 248-254
• 36 Bottcher C JF, van Gent CM, Pries C. A rapid and sensitive submicro phosphorus determination. Anal Chim Acta 1961; 24: 203-204
• 37 Fenton JW, Landis BH, Walz DA, Bing DH, Feinman RD, Zabinski MP, Sonder SA, Berliner LJ, Finlayson JS. Human thrombin: preparative evaluation, structural properties and enzymic specificity. In: The Chemistry and Physiology of the Human Plasma Proteins Bing DH. (ed.) pp 151-183 Pergamon Press; New York: 1979
• 38 Lollar P, Knutson GJ, Fass DN. Stabilization of thrombin-activated porcine factor VIII:C by factor IXa and phospholipid. Blood 1984; 63: 1303-1308
• 39 Stern DM, Drillings M, Kisiel W, Nawroth P, Nossel HL, LaGamma KS. Activation of factor IX bound to cultured bovine aortic endothelial cells. Proc Natl Acad Sci USA 1984; 81: 913-917
• 40 van Rijn J, Rosing J, van Dieijen G. Activity of human blood platelets in prothrombin and in factor X activation induced by ionophore A23187. Eur J Biochem 1983; 133: 1-10
• 41 Kao KJ, Pizzo SV, McKee PA. Demonstration and characterization of specific binding sites for factor VUI/von Willebrand factor on human platelets. J Clin Invest 1979; 63: 656-664
• 42 Andersson DO, Brown JE. Interaction of factor VUI-von Willebrand factor with phospholipid vesicles. Biochem J 1981; 200: 161-167
• 43 Brown JE, Baugh RF, Hougie C. Substrate inhibition of the intrinsic generation of activated factor X (Stuart factor). Thromb Res 1978; 13: 893-900
• 44 Jobin F, Esnouf MP. Studies on the formation of the prothrombin-converting complex. Biochem J 1967; 102: 666-674
• 45 Nesheim ME, Eid S, Mann KG. Assembly of the prothrombinase complex in the absence of prothrombin. J Biol Chem 1981; 256: 9874-9882
• 46 Lindhout T, Govers-Riemslag J WP, van de Waart P, Hemker HC, Rosing J. Factor Va-factor Xa interaction. Effects of phospholipid vesicles of varying composition. Biochemistry 1982; 21: 5494-5502
• 47 Nesheim ME, Tracy RP, Mann KG. “Clotspeed”, a mathematical simulation of the functional properties of prothrombinase. J Biol Chem 1984; 259: 1447-1453
• 48 Mertens K, Cupers R, van der Linden IK, Bertina RM. The functional defect of factor IX Eindhoven, a genetic variant of factor IX. Thromb Haemostas 1983; 50: 249 (Abstr.)
• 49 Mertens K, van Wijngaarden A, Bertina RM, Veltkamp JJ. The functional defect of factor VIII Leiden, a genetic variant of coagulation factor VIII. Thromb Haemostas 1985; 54: 650-653