Skip to main content
NCBI home page
Proceedings of the National Academy of Sciences of the United States of America logoLink to Proceedings of the National Academy of Sciences of the United States of America
. 1971 Jul;68(7):1653–1657. doi: 10.1073/pnas.68.7.1653

Mechanism of Action of Vitamin K: Evidence for the Conversion of a Precursor Protein to Prothrombin in the Rat

D V Shah 1, J W Suttie 1
PMCID: PMC389261  PMID: 5283959

Abstract

Previous studies have shown that the vitamin K-induced synthesis of prothrombin in a vitamin K-deficient rat is only slightly inhibited by cycloheximide treatment. Rat prothrombin has now been purified by disc electrophoresis after BaSO4 adsorption and citrate elution. When cycloheximide (5 mg/kg) was given to vitamin K-deficient rats 30 min before vitamin K, about 70% of the amount of prothrombin seen in rats not treated with cycloheximide was produced (two-stage assay), and the prothrombin band could be seen on the electrophoretic gels. However, if radioactive amino acids are administered to the rats after cycloheximide treatment, the newly formed prothrombin contains no radioactivity. The isolated prothrombin does contain radioactivity if the vitamin K-deficient rats are treated with vitamin K but no cycloheximide. When radioactive amino acids were given to deficient rats 1 hr before cycloheximide and vitamin K, radioactivity was found in prothrombin. These data suggest that, in the intact rat, the action of vitamin K is to convert a protein precursor with a short biological half life to prothrombin.

Keywords: phylloquinone, BaSO4 adsorption, polyacrylamide gel electrophoresis

Full text

PDF
1653

Images in this article

1654Image
on p.1654
1655Image
on p.1655
1656Image
on p.1656

Selected References

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

  1. ANDERSON G. F., BARNHART M. I. PROTHROMBIN SYNTHESIS IN THE DOG. Am J Physiol. 1964 May;206:929–938. doi: 10.1152/ajplegacy.1964.206.5.929. [DOI] [PubMed] [Google Scholar]
  2. Babior B. M., Kipnes R. S. Vitamin K dependent formation of factor VII by a cell-free system from rat liver. Biochemistry. 1970 Jun 23;9(13):2564–2569. doi: 10.1021/bi00815a002. [DOI] [PubMed] [Google Scholar]
  3. Babior B. M. The role of vitamin K in clotting factor synthesis. I. Evidence for the participation of vitamin K in the conversion of a polypeptide precursor to factor VII. Biochim Biophys Acta. 1966 Sep;123(3):606–610. [PubMed] [Google Scholar]
  4. DAVIS B. J. DISC ELECTROPHORESIS. II. METHOD AND APPLICATION TO HUMAN SERUM PROTEINS. Ann N Y Acad Sci. 1964 Dec 28;121:404–427. doi: 10.1111/j.1749-6632.1964.tb14213.x. [DOI] [PubMed] [Google Scholar]
  5. Dam H. International symposium on recent advances in research on vitamins K and related quinones (vitamins K, ubiquinones or coenzymes Q, plastoquinones) in honor of Professor Henrik Dam. Historical survey and introduction. Vitam Horm. 1966;24:295–306. [PubMed] [Google Scholar]
  6. Dulock M. A., Kolmen S. N. Influence of vitamin K on restoration of prothrombin complex proteins and fibrinogen in plasma of deplete dogs. Thromb Diath Haemorrh. 1968 Nov 15;20(1):136–143. [PubMed] [Google Scholar]
  7. GOLDSTEIN R., LE BOLLOC'H A., ALEXANDER B., ZONDERMAN E. Preparation and properties of prothrombin. J Biol Chem. 1959 Nov;234:2857–2866. [PubMed] [Google Scholar]
  8. Gorovsky M. A., Carlson K., Rosenbaum J. L. Simple method for quantitive densitometry of polyacrylamide gels using fast green. Anal Biochem. 1970 Jun;35(2):359–370. doi: 10.1016/0003-2697(70)90196-x. [DOI] [PubMed] [Google Scholar]
  9. Hemker H. C., Muller A. D. Kinetic aspects of the interaction of blood-clotting enzymes. VI. Localization of the site of blood-coagulation inhibition by the protein induced by vitamin K absence (PIVKA). Thromb Diath Haemorrh. 1968 Nov 15;20(1):78–87. [PubMed] [Google Scholar]
  10. Hill R. B., Gaetani S., Paolucci A. M., RamaRao P. B., Alden R., Ranhotra G. S., Shah D. V., Shah V. K., Johnson B. C. Vitamin K and biosynthesis of protein and prothrombin. J Biol Chem. 1968 Jul 25;243(14):3930–3939. [PubMed] [Google Scholar]
  11. Kipfer R. K., Olson R. E. Reversal by vitamin K of cycloheximide inhibited biosynthesis of prothrombin in the isolated perfused rat liver. Biochem Biophys Res Commun. 1970 Mar 27;38(6):1041–1048. doi: 10.1016/0006-291x(70)90344-x. [DOI] [PubMed] [Google Scholar]
  12. Li L. F., Kipfer R. K., Olson R. E. Immunochemical measurement of vitamin K-induced biosynthesis of prothrombin in the isolated perfused rat liver. Arch Biochem Biophys. 1970 Apr;137(2):494–499. doi: 10.1016/0003-9861(70)90467-4. [DOI] [PubMed] [Google Scholar]
  13. Li L. F., Olson R. E. Purification and properties of rat prothrombin. J Biol Chem. 1967 Dec 10;242(23):5611–5616. [PubMed] [Google Scholar]
  14. Losito R. Investigations into the presence of a competitive inhibitor (preprothrombin) in the plasma of chicks. Acta Chem Scand. 1965;19(9):2229–2234. doi: 10.3891/acta.chem.scand.19-2229. [DOI] [PubMed] [Google Scholar]
  15. Lowenthal J., Simmons E. L. Failure of actinomycin D to inhibit appearance of clotting activity by vitamin K in vitro. Experientia. 1967 Jun 15;23(6):421–422. doi: 10.1007/BF02142151. [DOI] [PubMed] [Google Scholar]
  16. MAMEESH M. S., JOHNSON B. C. Production of dietary vit. K deficiency in the rat. Proc Soc Exp Biol Med. 1959 Jul;101(3):467–468. doi: 10.3181/00379727-101-24982. [DOI] [PubMed] [Google Scholar]
  17. OLSON R. E. VITAMIN K INDUCED PROTHROMBIN FORMATION: ANTAGONISM BY ACTINOMYCIN D. Science. 1964 Aug 28;145(3635):926–928. doi: 10.1126/science.145.3635.926. [DOI] [PubMed] [Google Scholar]
  18. OYAMA V. I., EAGLE H. Measurement of cell growth in tissue culture with a phenol reagent (folin-ciocalteau). Proc Soc Exp Biol Med. 1956 Feb;91(2):305–307. doi: 10.3181/00379727-91-22245. [DOI] [PubMed] [Google Scholar]
  19. Olson J. P., Miller L. L., Troup S. B. Synthesis of clotting factors by the isolated perfused rat liver. J Clin Invest. 1966 May;45(5):690–701. doi: 10.1172/JCI105384. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Ranhotra G. S., Johnson B. C. Vitamin K and prothrombin formation. Life Sci. 1970 Jan 22;9(2):79–86. doi: 10.1016/0024-3205(70)90247-x. [DOI] [PubMed] [Google Scholar]
  21. Suttie J. W. Control of clotting factor biosynthesis by vitamin K. Fed Proc. 1969 Sep-Oct;28(5):1696–1701. [PubMed] [Google Scholar]
  22. Suttie J. W. The effect of cycloheximide administration on vitamin K-stimulated prothrombin formation. Arch Biochem Biophys. 1970 Dec;141(2):571–578. doi: 10.1016/0003-9861(70)90176-1. [DOI] [PubMed] [Google Scholar]
  23. Tishler P. V., Epstein C. J. A convenient method of preparing polyacrylamide gels for liquid scintillation spectrometry. Anal Biochem. 1968 Jan;22(1):89–98. doi: 10.1016/0003-2697(68)90262-5. [DOI] [PubMed] [Google Scholar]
  24. WETTSTEIN F. O., NOLL H., PENMAN S. EFFECT OF CYCLOHEXIMIDE ON RIBOSOMAL AGGREGATES ENGAGED IN PROTEIN SYNTHESIS IN VITRO. Biochim Biophys Acta. 1964 Jul 22;87:525–528. doi: 10.1016/0926-6550(64)90131-8. [DOI] [PubMed] [Google Scholar]
  25. WILLIAMS D. E., REISFELD R. A. DISC ELECTROPHORESIS IN POLYACRYLAMIDE GELS: EXTENSION TO NEW CONDITIONS OF PH AND BUFFER. Ann N Y Acad Sci. 1964 Dec 28;121:373–381. doi: 10.1111/j.1749-6632.1964.tb14210.x. [DOI] [PubMed] [Google Scholar]

Articles from Proceedings of the National Academy of Sciences of the United States of America are provided here courtesy of National Academy of Sciences

RESOURCES