Download PDF
Thromb Haemost 1976; 35(03): 576-585
DOI: 10.1055/s-0038-1648000
Original Article
Schattauer GmbH

Purification and Properties of Human Factor IXa

Katalin Váradi
1   Department of Blood Coagulation, National Institute of Haematology and Blood Transfusion, Budapest, Hungary
,
Susan Elödi
1   Department of Blood Coagulation, National Institute of Haematology and Blood Transfusion, Budapest, Hungary
› Author Affiliations
Further Information

Publication History

Received: 23 March 1975

Accepted: 06 September 1975

Publication Date:
02 July 2018 (online)

    Permissions and Reprints

Summary

Human factor IXa was purified 5,000-fold from serum by ion exchange chromatography. The preparation was free from other clotting factors. Both pH sensitivity and heat stability of purified factor IXa appeared to be different from those of factor IX in the plasma. The molecular weight of human factor IXa is 80,000 as estimated from gel-filtration experiments. Modification of seryl or histidyl side chains abolished the activity of factor IXa.

 
  • 1 Aronson D. L. 1974; Purification and properties of normal and abnormal factor IX. Thrombosis et Diathesis Haemorrhagica, Supplementum 59: 11.
    PubMedGoogle Scholar
  • 2 Biggs R. (ed.) 1972. Human Blood Coagulation, Haemostasis and Thrombosis. Blackwell Scientific Publications; Oxford:
    Google Scholar
  • 3 Biggs R, Mac Farlane R. G, Denson K. W. E, and Ash B. J. 1965; The interaction of factors VIII and IX. British Journal of Haematology 11: 276.
    CrossrefPubMedGoogle Scholar
  • 4 Chuang T. F, Sargeant R, and Hougie C. 1972; The intrinsic activation of factor X in blood coagulation. Biochimica, Biophysica Acta 273: 287.
    CrossrefPubMedGoogle Scholar
  • 5 Denson K. W. E. 1967. The Use of Antibodies in the Study of Blood Coagulation. Blackwell Scientific Publications; Oxford:
    Google Scholar
  • 6 Enfield D. L, Ericsson L. H, Fujikawa K, Titani K, Walsh K. A, and Neurath H. 1974; Bovine factor IX (Christmas factor). Further evidence of homology with factor X (Stuart factor) and prothrombin. FEBS Letters 47: 132.
    CrossrefPubMedGoogle Scholar
  • 7 Fujikawa K, Coan M. H, Enfield D. L, Titani K, Ericsson L. H, and Davie E. W. 1974; a A comparison of bovine prothrombin, factor IX (Christmas factor) and factor X (Stuart factor). Proceedings of the National Academy of Sciences, USA 71: A11.
    CrossrefPubMedGoogle Scholar
  • 8 Fujikawa K, Legaz M. E, Kato H, and Davie E. W. 1974; b The mechanism of activation bovine factor IX (Christmas factor) by bovine factor XIa (activated plasma thromboplastin antecedent). Biochemistry 13: 4508.
    CrossrefPubMedGoogle Scholar
  • 9 Hemker H. C, and de Graaf J. S. 1973 Les serine-esterases du systeme de la coagulation. Pathologie-Biologie. 21, Supplement, 5.
    PubMedGoogle Scholar
  • 10 Hemker H. C, and Kahn M. J. P. 1967; Reaction sequence of blood coagulation. Nature 215: 1201.
    CrossrefPubMedGoogle Scholar
  • 11 Hemker H. C, Kahn M. J. P, and Devilee P. P. 1970; The adsorption of coagulation factors onto phospholipids. Its role in the reaction mechanism of blood coagulation. Thrombosis et Diathesis Haemorrhagica 24: 214.
    PubMedGoogle Scholar
  • 12 Hershgold E. J, Davison A. M, and Janszen M. E. 1969; Human factor VIII (AHG), a lipoprotein: activation and inactivation by phospholipases. Federation Proceedings 28: 639.
    PubMedGoogle Scholar
  • 13 Hougie C. 1962; A simple assay method for factor X (Stuart-Prower factor). Proceedings of the Society for Experimental Biology and Medicine 109: 754.
    PubMedGoogle Scholar
  • 14 Hougie C, Denson K. W. E, and Biggs R. 1967; A study of the reaction product of factor VIII and factor IX by gel-filtration. Thrombosis et Diathesis Haemorrhagica 18: 211.
    PubMedGoogle Scholar
  • 15 Lowry O. H, Rosebrough N. J, Farr A. L, and Randall R. J. 1951; Protein measurement with the folin-phenol reagent. Journal of Biological Chemistry 193: 265.
    PubMedGoogle Scholar
  • 16 Melchior W. B, and Fahrney D. 1970; Ethoxyformylation of proteins. Reaction of ethoxyformic anhydride with α-chymotrypsin, pepsin and pancreatic ribonuclease at pH 4. Biochemistry 9: 251.
    CrossrefPubMedGoogle Scholar
  • 17 Mertz E. T, and Owen C. A. 1940; Imidazole buffer: its use in blood clotting studies. Proceedings of the Society for Experimental Biology and Medicine 43: 204.
    PubMedGoogle Scholar
  • 18 Norman P. S. 1957; Studies of the plasmin system. I. Measurement of human and animal plasminogen. Measurement of an activator in human serum. Journal of Experimental Medicine 106: 423.
    CrossrefPubMedGoogle Scholar
  • 19 Ovádi J, Libor S, and Elodi P. 1967; Spectrophotometric determination of histidine in proteins with diethyl-pyrocarbonate. Acta Biochimica et Biophysica Academia Scientiarium, Hungaricae 2: 455.
    PubMedGoogle Scholar
  • 20 Østerud B, and Schiffman S. 1972; Gelfiltration properties of factors II, VII, IX, IX a and X. Thrombosis et Diathesis Haemorrhagica 28: 317.
    PubMedGoogle Scholar
  • 21 Radcliffe R. D, and Barton P. G. 1973; Comparison of the molecular forms of activated bovine factor X. Journal of Biological Chemistry 248: 6788.
    PubMedGoogle Scholar
  • 22 Schmer G, Kirby E. P, Teller D. C, and Davie E. W. 1972; The isolation and characterization of bovine factor VIII (antihaemophilic factor). Journal of Biological Chemistry 247: 2512.
    PubMedGoogle Scholar
  • 23 Somer J. B, and Castaldi P. A. 1970; Coagulation factor IX in normal and haemophilia B plasma. British Journal of Haematology 18: 147.
    CrossrefPubMedGoogle Scholar
  • 24 Thomson J. M. 1970. A Practical Guide to Blood Coagulation and Haemostasis. J. A. Churchill; London:
    Google Scholar
  • 25 Titani K, Hermodson M. A, Fujikawa K, Ericsson L. H, Walsh K. A, Neurath H, and Davie E. W. 1972; Bovine factor Xa (activated Stuart factor). Evidence of homology with mammalian serine proteases. Biochemistry 11: 4899.
    CrossrefPubMedGoogle Scholar