Thromb Haemost 1979; 42(03): 838-847
DOI: 10.1055/s-0038-1666932
Original Article
Schattauer GmbH Stuttgart

Inactivation of Factor VIII by a Mechanism Independent of the Generation of Thrombin[*]

D C Triantaphyllopoulos
The American Red Cross Blood Research Laboratory, Bethesda, Md. and Dept. of Physiology and Biophysics, Georgetown University School of Medicine, Washington, D. C., USA
› Author Affiliations
Further Information

Publication History

Received 23 September 1978

Accepted 14 November 1978

Publication Date:
23 August 2018 (online)

Summary

Thrombin first activates and then inactivates factor VIII and for this reason thrombin has been considered responsible for the inactivation of factor VIII which occurs during clotting. Experiments described in this paper indicated that the activity of factor VIII is not reduced in factor IX or factor X deficient sera, while on the other hand this factor becomes inactivated in blood anticoagulated with high concentrations of hirudin which inhibit thrombin activity completely. This suggests that some other factor, besides thrombin, which is generated only in trace amounts in factor IX or factor X deficient plasmas, is also able to inactivate factor VIII. Purified factor X activated with insolubilized trypsin was added to purified preparations of factor VIII, which were free of both fibrinogen and prothrombin. Factor Xa was allowed to act for 5-60 minutes and then inactivated with phenylmethanesul- fonyl fluoride. Depending on the duration of the action of factor X a partial or complete inactivation of factor VIII was observed. This inactivation was also observed in the presence of hirudin, thus excluding the possibility that the effect was due to contamination with trace amounts of thrombin.

* Presented in part at the 61st Annual Meeting of the Federation of the American Societies for Experimental Biology and at the 20th Annual Meeting of the American Society of Hematology.


 
  • References

  • 1 Biggs R. 1972. Intrinsic prothrombin activation. In Biggs R. (ed.) Human Blood Coagulation, Haemostasis and Thrombosis. Blackwell Scientific Publications; Oxford: p 64
  • 2 Biggs R, Douglas AS. 1953; The thromboplastin generation test. Journal of Clinical Pathology 6: 23
  • 3 Biggs R, MacFarlane RG, Denson KW E, Ash BJ. 1965; Thrombin and the interaction of factors VIII and IX. British Journal of Haematolgy 11: 276
  • 4 Davie EW, Fujikawa K. 1975; Basic mechanisms in blood coagulation. Annual Revue of Biochemistry 44: 799
  • 5 Davie EW, Fujikawa K, Kato H, Legaz ME. 1975; Properties of bovine factor IX (Christmas factor). Annals of the New York Academy of Science 240: 24
  • 6 Esnouf MP, Lloyd PH, Jesty J. 1973; A method for the simultaneous isolation of factor X and prothrombin from bovine plasma. Biochemical Journal 131: 781
  • 7 Irwin JR, Seegers WH, Andary TJ, Fekete LF, Nova E. 1975; Blood coagulation as a cybernetic system: Control of autoprothrombin C (factor Xa) formation. Thrombosis Research 6: 431
  • 8 James HL, Wickerhauser M. 1972; Development of large-scale fractionation methods. III Preparation of factor VIII concentrate of intermediate purity. Vox Sanguinis 23: 402
  • 9 MacFarlane RG, Ash BJ. 1968; The activation and consumption of factor X in recalcified plasma. British Journal of Haematology 10: 217
  • 10 Markwardt F, Lindmann H. 1971; Blutgerinnunsghemmende Proteine, Peptide und Aminosaurederivative. Handbuch der Experimentellen Pharmakologie 27: 76
  • 11 Osterud B, Rapaport SI, Schiffman S, Chong MM Y. 1971; Formation of intrinsic factor X-activator activity with special reference to the role of thrombin. British Journal of Haematology 21: 643
  • 12 Penick GD. 1957; Some factors that influence utilization of antihemophilic activity during clotting. Proceedings of the Society for Experimental Biology and Medicine 96: 277
  • 13 Reno RS, Seegers WH. 1967; The activation and consumption of factor X in recalcified plasma. Thrombosis et Diathesis flaemorrhagica 18: 198
  • 14 Seegers WH, Marciniak E, Mccoy L. 1969; Note on the inactivation of autoprothrombin C with diisopropylfluorophosphate or phenylmethanesulfonyl fluoride. Thrombosis et Diathesis Haemorrhagica 25: 32
  • 15 Triantaphyllopoulos DC. 1958; Anticoagulant effect of incubated fibrinogen. Canadian Journal of Biochemistry and Physiology 36: 249
  • 16 Triantaphyllopoulos DC. 1960; Effects of intravenous injections of the anticoagulant fraction of incubated fibrinogen on blood coagulation. Candian Journal of Biochemistry and Physiology 38: 909
  • 17 Triantaphyllopoulos DC. 1974; Factor XIII consumption as an indicator of thrombin generation. Journal of Laboratory and Clinical Medicine 16: 589
  • 18 Triantaphyllopoulos DC. 1976. Physiological effects of the plasminolytic derivatives of fibrinogen. In Jamieson GA, Greenwalt TJ. eds Trace Components of Plasma: Isolation and Clinical Significance. Alan R. Liss, Inc; New York: p 121
  • 19 Triantaphyllopoulos DC, Chen C, Triantaphyllopoulos E. 1969; Nature of the inhibition of prothrombin consumption by lysed fibrinogen. British Journal of Haematology 16: 589
  • 20 Triantaphyllopoulos DC, Ryan LT. 1976; Fibrinogen and its derivatives: Cofactors in the intrinsic generation of thrombin. Thrombosis and Haemostasis 35: 305
  • 21 Ware AG, Seegers WH. 1949; Two stage procedure for the quantitative determination of prothrombin concentration. American Journal of Clinical Pathology 19: 471