Skip to main content
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
. 1975 Jan;72(1):131–135. doi: 10.1073/pnas.72.1.131

Mitogenic activity of blood components. I. Thrombin and prothrombin.

L B Chen, J M Buchanan
PMCID: PMC432255  PMID: 1054489

Abstract

When added to a culture medium of resting confluent chick embryo fibroblasts in the absence of serum, thrombin (EC 3.4.21.5) is able to stimulate DNA synthesis 12 hr later and to cause a substantial increase in cell number over a period of 4 days. As compared to thrombin, prothrombin exhibits low mitogenic activity. However, in the presence of purified Factor Xa (EC 3.4.21.6) and Factor V, prothrombin is converted to thrombin by "thromboplastin activity" supplied by the fibroblasts. Prothrombin, either purified or as a constituent of plasma or serum, may thus be considered to be a reservoir of mitogenic activity in tissue culture unless antithrombin is present in the culture medium in amounts sufficient to neutralize the thrombin formed. By use of a specific inhibitor of proteases, and by separation of prothrombin by absorption on BaSO4, we estimate that the potential mitogenic activity of prothrombin is approximately 30-50% of the total activity that can be obtained by treatment of fibrinogen-free plasma with thromboplastin. In addition to its mitogenic activity, thrombin can also stimulate the migration of cells. These experiments with thrombin illustrate that well-characterized proteases of blood can act as potent mitogens and suggest that they may play a role in the process of wound healing.

Full text

PDF
133

Selected References

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

  1. Balk S. D., Whitfield J. F., Youdale T., Braun A. C. Roles of calcium, serum, plasma, and folic acid in the control of proliferation of normal and Rous sarcoma virus-infected chicken fibroblasts. Proc Natl Acad Sci U S A. 1973 Mar;70(3):675–679. doi: 10.1073/pnas.70.3.675. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Baughman D. J., Waugh D. F. Bovine thrombin. Purification and certain properties. J Biol Chem. 1967 Nov 25;242(22):5252–5259. [PubMed] [Google Scholar]
  3. Burger M. M. Proteolytic enzymes initiating cell division and escape from contact inhibition of growth. Nature. 1970 Jul 11;227(5254):170–171. doi: 10.1038/227170a0. [DOI] [PubMed] [Google Scholar]
  4. Bürk R. R. A factor from a transformed cell line that affects cell migration. Proc Natl Acad Sci U S A. 1973 Feb;70(2):369–372. doi: 10.1073/pnas.70.2.369. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Dulak N. C., Temin H. M. Multiplication-stimulating activity for chicken embryo fibroblasts from rat liver cell conditioned medium: a family of small polypeptides. J Cell Physiol. 1973 Apr;81(2):161–170. doi: 10.1002/jcp.1040810205. [DOI] [PubMed] [Google Scholar]
  6. Esnouf M. P., Jobin F. The isolation of factor V from bovine plasma. Biochem J. 1967 Mar;102(3):660–665. doi: 10.1042/bj1020660. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Esnouf M. P., Macfarlane R. G. Enzymology and the blood clotting mechanism. Adv Enzymol Relat Areas Mol Biol. 1968;30:255–315. doi: 10.1002/9780470122754.ch6. [DOI] [PubMed] [Google Scholar]
  8. Fujikawa K., Legaz M. E., Davie E. W. Bovine factor X 1 (Stuart factor). Mechanism of activation by protein from Russell's viper venom. Biochemistry. 1972 Dec 19;11(26):4892–4899. doi: 10.1021/bi00776a003. [DOI] [PubMed] [Google Scholar]
  9. Fujikawa K., Legaz M. E., Davie E. W. Bovine factors X 1 and X 2 (Stuart factor). Isolation and characterization. Biochemistry. 1972 Dec 19;11(26):4882–4891. doi: 10.1021/bi00776a002. [DOI] [PubMed] [Google Scholar]
  10. 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]
  11. Gospodarowicz D. Localisation of a fibroblast growth factor and its effect alone and with hydrocortisone on 3T3 cell growth. Nature. 1974 May 10;249(453):123–127. doi: 10.1038/249123a0. [DOI] [PubMed] [Google Scholar]
  12. Holley R. W., Kiernan J. A. "Contact inhibition" of cell division in 3T3 cells. Proc Natl Acad Sci U S A. 1968 May;60(1):300–304. doi: 10.1073/pnas.60.1.300. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Hollwy R. W., Kiernan J. A. Control of the initiation of DNA synthesis in 3T3 cells: serum factors. Proc Natl Acad Sci U S A. 1974 Jul;71(7):2908–2911. doi: 10.1073/pnas.71.7.2908. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Marchalonis J. J. An enzymic method for the trace iodination of immunoglobulins and other proteins. Biochem J. 1969 Jun;113(2):299–305. doi: 10.1042/bj1130299. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Rein A., Rubin H. Effects of local cell concentrations upon the growth of chick embryo cells in tissue culture. Exp Cell Res. 1968 Mar;49(3):666–678. doi: 10.1016/0014-4827(68)90213-9. [DOI] [PubMed] [Google Scholar]
  16. Rosenberg R. D., Waugh D. F. Multiple bovine thrombin components. J Biol Chem. 1970 Oct 10;245(19):5049–5056. [PubMed] [Google Scholar]
  17. Sefton B. M., Rubin H. Release from density dependent growth inhibition by proteolytic enzymes. Nature. 1970 Aug 22;227(5260):843–845. doi: 10.1038/227843a0. [DOI] [PubMed] [Google Scholar]
  18. Temin H. M. Studies on carcinogenesis by avian sarcoma viruses. VI. Differential multiplication of uninfected and of converted cells in response to insulin. J Cell Physiol. 1967 Jun;69(3):377–384. doi: 10.1002/jcp.1040690314. [DOI] [PubMed] [Google Scholar]
  19. Todaro G., Matsuya Y., Bloom S., Robbins A., Green H. Stimulation of RNA synthesis and cell division in resting cells by a factor present in serum. Wistar Inst Symp Monogr. 1967;7:87–101. [PubMed] [Google Scholar]
  20. 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]
  21. Zacharski L. R., McIntyre O. R. Membrane-mediated synthesis of tissue factor (thromboplastin) in cultured fibroblasts. Blood. 1973 May;41(5):679–685. [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