Thromb Haemost 1976; 36(01): 104-114
DOI: 10.1055/s-0038-1648014
Original Article
Schattauer GmbH

The Activation of Human Factor X in Sodium Citrate: the Role of Factor VII[*]

D. L Aronson
1   Bureau of Biologics, Food and Drug Administration, Bethesda, MD 20014, USA
,
A. J Mustafa
1   Bureau of Biologics, Food and Drug Administration, Bethesda, MD 20014, USA
› Author Affiliations
Further Information

Publication History

Received 26 September 1975

Accepted 04 January 1976

Publication Date:
03 July 2018 (online)

Summary

Human factor X was purified by several different procedures yielding products which had varying amounts of factor VII and factor IX. Treatment with CHC13 during the fractionation of the factor X removed 95% of the factor VII and factor IX activity and the resulting factor X activated more slowly when incubated in 25% sodium citrate. Removal of residual factor VII by DEAE cellulose chromatography yielded a factor X which activated still more slowly and less completely. When the factor VII, removed by chromatography, was added to the chromatographed factor X, the ability to be activated in 25% sodium citrate was restored. Confirmatory evidence for the role of factor VII in this reaction was the inhibition of the conversion of the factor X by both DFP and SBTI.

* This work was presented at the American Hematology Society’s 1971 meeting.


 
  • References

  • 1 Aronson D. L, Dodds W. J, Mustafa A. J. 1972; A Quantitative Two-Stage Assay for Factor IX (Christmas Factor) Using Plasma from Dogs with Christmas Disease. Thrombosis et Diathesis Haemorrhagica 27: 529.
  • 2 Aronson D. L, Mustafa A. J. 1971; Two Forms of the Human Prothrombin Converting Enzyme (Active Factor X). Proceedings of the Society for Experimental Biology and Medicine 137: 1262.
  • 3 Aronson D. L, Mustafa A. J, Mushinski J. F. 1969; Purification of Human Factor X and Comparison of Peptide Maps of Human Factor X and Prothrombin. Biochemica et Biophysica Acta 188: 25.
  • 4 Biggs R, Macfarlane R. G, Denson K. W. E, Ash B. J. 1964; The Interaction of Factors VIII and IX. British Journal of Haematology 10: 530.
  • 5 Davis B. J. 1964; Disc Electrophoresis II Method and Application Human Serum Proteins. Annals of the Academy of Science 121: 404.
  • 6 Denson K. W. E. 1961; The Specific Assay of Prower-Stuart Factor and Factor VIL Acta Haematologica. 25: 105.
  • 7 Denson K. W. E. 1967. The Use of Antibodies in the Study of Blood Coagulation, Chapter 2. Black-well Scientific Publication; London, England.:
  • 8 Esnouf M. P, Williams W. J. 1972; The Isolation and Purification of a bovine-plasma protein which is a substrate for the coagulant fraction of Russell’s-viper venom. Biochemical Journal 84: 62.
  • 9 Hoag M. S, Johnson F. J, Robinson J. A, Aggeler P.M. 1969; Treatment of Hemophilia B with a new clotting-factor concentrate. New England Journal of Medicine 280: 581.
  • 10 Hougie C, Bunting R. 1959; Separation of Factor VII and Stuart Factor by DEAE cellulose. Thrombosis et Diathesis Haemorrhagica 4 (Suppl. 01) 40.
  • 11 Hougie C, Denson K. W. W, Biggs R. 1967; A Study of the Reaction Product of Factor VIII and Factor IX by Gel Filtration. Thrombosis et Diathesis Haemorrhagica 18: 211.
  • 12 Jackson C. M, Hanahan D. M. 1968; Studies on bovine Factor XII. Characterization of Purified Factor X. Observations on some alterations in zone electrophoretic and chromatographic behavior occurring during purification. Biochemistry 7: 4506.
  • 13 Jesty J, Nemerson Y. 1974; Purification of Factor VII from Bovine Plasma. Journal of Biological Chemistry 249: 509.
  • 14 Jesty J, Spencer A. K, Nemerson Y. 1964; The Mechanism of Activation of Factor X. Journal of Biological Chemistry 249: 5614.
  • 15 Lanchantin G. F, Friedman J. A, Hart D. W, Aronson D. L. 1969; Studies on the mechanism of human prothrombin activation by Factor X. Federation Proceedings 28: 639.
  • 16 Leveson J. E, Esnouf M. P. 1969; The inhibition of activated Factor X with diisopropryl-fluorophosphate. British Journal of Haematology 17: 173.
  • 17 Lundblad R. L, Davie E. W. 1965; The Activation of Stuart Factor (Factor X) by Activated Antihemophilic Factor (Activated Factor VIII). Biochemistry 4: 113.
  • 18 Macfarlane R. G, Ash B. J. 1964; The Activation and Consumption of Factor X in Recalcified Plasma. British Journal of Haematology 10: 217.
  • 19 Miller K. D, Van Vunakis H. 1956; The effect of Diisopropylfluorophosphate on the Proteinase and Esterase Activities of Thrombin and on Prothrombin and Its Activators. Journal of Biological Chemistry 223: 277.
  • 20 Milstone J. H. 1948; Three-stage analysis of blood coagulation. Journal of General Physiology 32: 301.
  • 21 Milstone J. H. 1951; Fractionation of Plasma Globulin for Prothrombin, Thrombokinase and Accessory Thromboplastin. Journal of General Physiology 35: 67.
  • 22 Milstone J. H. 1955; Effect of Blood Thrombokinase, as Influenced by Soy Bean Trypsin Inhibitor, Ultracentrifugation and Accessory Factors. Journal of General Physiology 38: 757.
  • 23 Milstone J. H, Oulianoff N, Saxton T. R, Milstone V. K. 1971; Partial and Selective Inactivation of Thrombokinase by Chymotrypsin. Yale Journal of Biology and Medicine 43: 223.
  • 24 Nemerson Y. 1966; The Reaction between Bovine Brain Tissue and Factors VII and X. Biochemistry 5: 601.
  • 25 Nemerson Y, Spaet T. H. 1964; The Activation of Factor X by Extracts of Rabbit Brain. Blood 23: 657.
  • 26 Osterud B, Berre A, Otnaess A. B, Bjorklid E, Prydz H. 1972; Activation of the coagulation Factor VII by tissue thromboplastin and Calcium. Biochemistry 11: 2853.
  • 27 Osterud B, Rapaport S. I. 1970; Synthesis of intrinsic Factor X activator. Inhibition of the function of formed activator by antibodies to Factor VII and to Factor IX. Biochemistry 9: 1854.
  • 28 Papahadjopoulos D, Yin E. T, Hanahan D. J. 1964; Purification and Properties of Bovine Factor X: Molecular Changes during Activation. Biochemistry 3: 1931.
  • 29 Pechet L, Smith J. A. 1970; The separation of clotting factors II (Prothrombin) and IX Please Thromboplastin component by isoelectric focusing. Biochemica et Biophysica Acta 200: 475.
  • 30 Radcliffe R. D, Barton P. G. 1973; Comparisons of the Molecular Forms of Activated Bovine Factor X. Journal of Biological Chemistry 248: 6788.
  • 31 Radcliffe and Nemerson. 1974; Activation and Control of Factor VII by Activated Factor X and Thrombin. Journal of Biological Chemistry 250: 388.
  • 32 Seegers W. H, Heene D. L, Marciniak E. 1966; Activation of Purified Prothrombin in Ammonium Sulfate Solutions: Purification of Authoprothrombin C. Thrombosis et Diathesis Haemor-rhagica 15: 1.
  • 33 Somer J. B, Castaldi P. A. 1970; Coagulation Factor IX in Normal and Hemophilia B plasma. British Journal of Haematology 18: 147.
  • 34 Spaet T. H. 1964; Nature and action of intermediate product I. Federation Proceedings 23: 757.
  • 35 Spaet T. H, Cintron J. 1963; Pathways to blood coagulation Product I formation. Blood 21: 745.
  • 36 Tagnon H. J, Soulier J. P. 1946; Anticoagulant Activity of the Trypsin Inhibitor from Soya Bean Flour. Proceedings of Society for Experimental Biology and Medicine 61: 440.