Thromb Haemost 1972; 28(02): 317-324
DOI: 10.1055/s-0038-1649013
Original Article
Schattauer GmbH

Gel Filtration Properties of Factors II, VII, IX, IXa, and X

Bjarne Østerud*
1   Departments of Medicine and Biochemistry, University of Southern California School of Medicine, Los Angeles, California, 90033
,
Sandra Schiffman
1   Departments of Medicine and Biochemistry, University of Southern California School of Medicine, Los Angeles, California, 90033
› Author Affiliations
Further Information

Publication History

Publication Date:
24 July 2018 (online)

Summary

The following gel filtration properties of the vitamin ढ dependent clotting factors have been established :

1. The elution patterns of factors II, VII, IX, and X in 0.5 M NaCl are the same in factor XII deficient native plasma, anticoagulated plasma, and BaSO4 eluates.

2. In 0.01 M Tris buffer pH 7.3 or 0.5 M NaCl, plasma factors II, IX, and X elute together; factor VII élûtes later as a smaller molecule. All 4 plasma activities elute as larger molecules in 0.01 M Tris buffer and smaller ones in 0.5 M NaCl.

3. In 0.01 M Tris - 0.005 M EDTA peak factor II activity elutes first, followed by factor IX, factor X, and then factor VII.

4. Activation decreases the apparent size of factor IX.

* Present address: Department of Microbiology Dental Faculty, University of Oslo, Oslo, Norway.


 
  • References

  • 1 Andrews P. 1970; Estimation of molecular size and molecular weights of biological compounds by gel filtration. Methods of Biochemical Analysis 18: 1.
  • 2 Derleth Th. R, and Penner J. A. 1970; Human prothrombin. Chromatographic separation from normal and deficient plasmas. Thrombosis et Diathesis Haemorrhagica 24: 311.
  • 3 Deutsch E, Lechner K, and Schmer G. 1966; On the purification of Factor VII from bovine plasma. Thrombosis et Diathesis Haemorrhagica Suppl 20: 275.
  • 4 Hjort P, Rapaport S. I, and Owren P. A. 1955; A simple, specific one-stage prothrombin assay using Russell’s viper venom in cephalin suspension. Journal of Laboratory and Clinical Medicine 46: 89.
  • 5 Hjort P. F. 1957; Intermediate reactions in the coagulation of blood with tissue thromboplastin. Scandinavian Journal of Clinical and Laboratory Investigation 9 (27) 44.
  • 6 Hougie C. 1962; A simple assay method for Factor X (Stuart-Prower Factor). Proceedings of the Society for Experimental Biology and Medicine 109: 75a.
  • 7 Jackson C. M, and Hanahan D. J. 1968; Studies on bovine factor X. II. Characterization of purified factor X. Observations on some alternations in zone electrophoretic and chromatographic behavior occurring during purification. Biochemistry 7: 4506.
  • 8 Kingdon H. 1969; Evolution of enzyme activities and fibrin formation after contact of plasma with glass. Journal of Biomedical Materials Research 3: 25.
  • 9 Lanchantin G. F, Friedmann J. A, and Hart D. W. 1965; The conversion of human prothrombin to thrombin by sodium citrate. Analysis of the activation mixture. Journal of Biological Chemistry 240: 3276.
  • 10 Lanchantin G. F, Friedmann J. A, and Hart D. W. 1967; Esterase and clotting activities derived from citrate activation of human prothrombin. Journal of Biological Chemistry 242: 2491.
  • 11 Lewis J. H. 1964; Separation and molecular weight estimation of coagulation and fibrinolytic proteins by Sephadex gel filtration. Proceedings of the Society for Experimental Biology and Medicine 116: 120.
  • 12 Østerud B, Rapaport S. I, Schiffman S, and Chong M. M. Y. 1971; Formation of intrinsic factor-X-activator activity, with special reference to the role of thrombin. British Journal of Haematology 21: 643.
  • 13 Prydz H. 1963; Studies on proconvertin. I. Gel Filtration. Scandinavian Journal of Clinical and Laboratory Investigation 15: 450.
  • 14 Prydz H. 1965; Some characteristics of purified factor VII preparations. Scandinavian Journal of Clinical and Laboratory Investigation 17 (84) 78.
  • 15 Prydz H. 1969. Factor VII and tissue thromboplastin. In: Hemker H. C, Loeliger E. A, and Veltkamp J. J. (eds.) Human Blood Coagulation. Leiden University Press; 83.
  • 16 Rasche H, Hiemeyer V, and Melee A. 1970; Das Verhalten des Blutgerinnungsfaktors X (Stuart-Prower) aus Plasma und Serum bei der Gelfiltration an Sephadex G-200. Thrombosis et Diathesis Haemorrhagica 23: 405.
  • 17 Schiffman S, Rapaport S. I, and Patch M. J. 1963; The identification and synthesis of activated plasma thromboplastin component (PTC). Blood 22: 733.
  • 18 Schiffman S, Rapaport S. I, and Patch M. J. 1965; Starch block electrophoresis of plasma and serum clotting factors. Separation of activated PTC (PTC). Blood 25: 724.
  • 19 Shapiro S. S, and Waugh D. F. 1966; The purification of human prothrombin. Thrombosis et Diathesis Haemorrhagica 16: 469.
  • 20 Swart A. C. W, and Hemker H. C. 1970; Separation of blood coagulation factors II, VII, IX, and X by gel filtration in the presence of Dextran blue. Biochimica et Biophysica Acta 222: 692.
  • 21 Tishkoff G, IL Williams L. C, and Brown D. M. 1968; Preparation of highly purified prothrombin complex. I. Crystalization, biological activity, and molecular properties. Journal of Biological Chemistry 243: 4151.