Skip to main content
PMC home page
Biochemical Journal logoLink to Biochemical Journal
. 1975 Mar;145(3):469–474. doi: 10.1042/bj1450469

Purification and some characteristics of the coagulation factor IX from human plasma.

B Osterud, R Flengsrud
PMCID: PMC1165246  PMID: 1171684

Abstract

Non-activated coagulation factor IX was purified approx. 10,000-fold from human plasma. The final product was electrophoretically homogeneous and comprised a tingle polypeptide chain with a molecular weight of about 70,000 and a pI of 4.3-4.45. The N-terminal amino acid was glycine. The amino acid and the carbohydrate contents were analysed and a monospecific antiserum to the factor was raised in rabbits.

Full text

PDF
473

Images in this article

472-1PLATE 1
on p.472-1
472-2PLATE 2
on p.472-2

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Barton P. G. Sequence theories of blood coagulation re-evaluated with reference to lipid-protein interactions. Nature. 1967 Sep 30;215(5109):1508–1509. doi: 10.1038/2151508a0. [DOI] [PubMed] [Google Scholar]
  2. Bjorklid E., Storm E., Prydz H. The protein component of human brain thromboplastin. Biochem Biophys Res Commun. 1973 Dec 10;55(3):969–976. doi: 10.1016/0006-291x(73)91237-0. [DOI] [PubMed] [Google Scholar]
  3. Böhlen P., Stein S., Dairman W., Udenfriend S. Fluorometric assay of proteins in the nanogram range. Arch Biochem Biophys. 1973 Mar;155(1):213–220. doi: 10.1016/s0003-9861(73)80023-2. [DOI] [PubMed] [Google Scholar]
  4. Catsimpoolas N. Micro isoelectric focusing in polyacrylamide gel columns. Anal Biochem. 1968 Dec;26(3):480–482. doi: 10.1016/0003-2697(68)90219-4. [DOI] [PubMed] [Google Scholar]
  5. Chandra S., Pechet L. The separation of clotting factors. II. Purification of human factor IX by isoelectric focusing. Biochim Biophys Acta. 1973 Dec 6;328(2):456–463. [PubMed] [Google Scholar]
  6. Cuatrecasas P. Protein purification by affinity chromatography. Derivatizations of agarose and polyacrylamide beads. J Biol Chem. 1970 Jun;245(12):3059–3065. [PubMed] [Google Scholar]
  7. Deyl Z., Rosmus J. Thin layer chromatography of Dansyl amino acid derivatives. J Chromatogr. 1965 Dec;20(3):514–520. doi: 10.1016/s0021-9673(01)97453-9. [DOI] [PubMed] [Google Scholar]
  8. Edelhoch H. Spectroscopic determination of tryptophan and tyrosine in proteins. Biochemistry. 1967 Jul;6(7):1948–1954. doi: 10.1021/bi00859a010. [DOI] [PubMed] [Google Scholar]
  9. Fujikawa K., Coan M. H., Enfield D. L., Titani K., Ericsson L. H., Davie E. W. A comparison of bovine prothrombin, factor IX (Christmas factor), and factor X (Stuart factor). Proc Natl Acad Sci U S A. 1974 Feb;71(2):427–430. doi: 10.1073/pnas.71.2.427. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Fujikawa K., Thompson A. R., Legaz M. E., Meyer R. G., Davie E. W. Isolation and characterization of bovine factor IX (Christmas factor). Biochemistry. 1973 Nov 20;12(24):4938–4945. doi: 10.1021/bi00748a019. [DOI] [PubMed] [Google Scholar]
  11. Gros C., Labouesse B. Study of the dansylation reaction of amino acids, peptides and proteins. Eur J Biochem. 1969 Feb;7(4):463–470. doi: 10.1111/j.1432-1033.1969.tb19632.x. [DOI] [PubMed] [Google Scholar]
  12. Howell R. M., Dupe R. J. Observations on factor X activity after adsorption to barium sulphate. Thromb Diath Haemorrh. 1972 Oct 31;28(2):306–316. [PubMed] [Google Scholar]
  13. LOWRY O. H., ROSEBROUGH N. J., FARR A. L., RANDALL R. J. Protein measurement with the Folin phenol reagent. J Biol Chem. 1951 Nov;193(1):265–275. [PubMed] [Google Scholar]
  14. Laemmli U. K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. doi: 10.1038/227680a0. [DOI] [PubMed] [Google Scholar]
  15. Neville D. M., Jr Molecular weight determination of protein-dodecyl sulfate complexes by gel electrophoresis in a discontinuous buffer system. J Biol Chem. 1971 Oct 25;246(20):6328–6334. [PubMed] [Google Scholar]
  16. ORNSTEIN L. DISC ELECTROPHORESIS. I. BACKGROUND AND THEORY. Ann N Y Acad Sci. 1964 Dec 28;121:321–349. doi: 10.1111/j.1749-6632.1964.tb14207.x. [DOI] [PubMed] [Google Scholar]
  17. Osterud B., Rapaport S. I., Schiffman S., Chong M. M. Formation of intrinsic factor-X activator activity, with special reference to the role of thrombin. Br J Haematol. 1971 Dec;21(6):643–660. doi: 10.1111/j.1365-2141.1971.tb02727.x. [DOI] [PubMed] [Google Scholar]
  18. Osterud B., Rapaport S. I. Synthesis of intrinsic factor X activator. Inhibition of the function of formed activator by antibodies to factor VIII and to factor IX. Biochemistry. 1970 Apr 14;9(8):1854–1861. doi: 10.1021/bi00810a028. [DOI] [PubMed] [Google Scholar]
  19. Osterud B., Schiffman S. Gel filtration properties of factors II, VII, IX, IX a , and X. Thromb Diath Haemorrh. 1972 Oct 31;28(2):317–324. [PubMed] [Google Scholar]
  20. Prydz H., Gladhaug A. A specific system for he estimation of prothrombin activity. Thromb Diath Haemorrh. 1970 Dec 31;24(3):601–602. [PubMed] [Google Scholar]
  21. SCHEIDEGGER J. J. Une micro-méthode de l'immuno-electrophorèse. Int Arch Allergy Appl Immunol. 1955;7(2):103–110. [PubMed] [Google Scholar]
  22. Skotland T., Holm T., Osterud B., Flengsrud R., Prydz H. The localization of a vitamin K-induced modification in an N-terminal fragment of human prothrombin. Biochem J. 1974 Oct;143(1):29–37. doi: 10.1042/bj1430029. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. WARREN L. The thiobarbituric acid assay of sialic acids. J Biol Chem. 1959 Aug;234(8):1971–1975. [PubMed] [Google Scholar]
  24. WINZLER R. J. Determination of serum glycoproteins. Methods Biochem Anal. 1955;2:279–311. doi: 10.1002/9780470110188.ch10. [DOI] [PubMed] [Google Scholar]
  25. Weber K., Osborn M. The reliability of molecular weight determinations by dodecyl sulfate-polyacrylamide gel electrophoresis. J Biol Chem. 1969 Aug 25;244(16):4406–4412. [PubMed] [Google Scholar]
  26. Woods K. R., Wang K. T. Separation of dansyl-amino acids by polyamide layer chromatography. Biochim Biophys Acta. 1967 Feb 21;133(2):369–370. doi: 10.1016/0005-2795(67)90078-5. [DOI] [PubMed] [Google Scholar]

Articles from Biochemical Journal are provided here courtesy of The Biochemical Society

RESOURCES