Skip to main content
PMC home page
The Journal of Clinical Investigation logoLink to The Journal of Clinical Investigation
. 1986 Jun;77(6):1909–1916. doi: 10.1172/JCI112519

Activation of endogenous factor V by a homocysteine-induced vascular endothelial cell activator.

G M Rodgers, W H Kane
PMCID: PMC370551  PMID: 3519683

Abstract

Vascular endothelium possesses multiple procoagulant properties, including synthesis and expression of Factor V. We studied the effects of homocysteine on the regulation of endothelial cell Factor V activity. Elevated levels of homocysteine are associated with the congenital thrombotic disorder homocystinuria. Treatment of cultured endothelial cells with 0.5-10 mM homocysteine had no effect on cell morphology, but did increase Factor V activity and prothrombin activation by Factor Xa. A radioimmunoassay for endothelial cell Factor V demonstrated that homocysteine treatment did not increase Factor V antigen levels. 125I-prothrombin was activated by treated endothelial cells and Factor Xa in the presence of thrombin inhibitors. Exogenous 125I-Factor V was cleaved by homocysteine-treated but not control endothelial cells. 125I-Factor V cleavage products distinct from those generated by thrombin and Factor Xa were identified. These data provide evidence for regulation of endothelial cell Factor V activity, and indicate that increased Factor V activity associated with homocysteine-treated vascular endothelium results primarily from induction of an activator of Factor V.

Full text

PDF

Images in this article

1913Image
on p.1913
1913Image
on p.1913

Selected References

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

  1. BACHMANN F., DUCKERT F., KOLLER F. The Stuart-Prower factor assay and its clinical significance. Thromb Diath Haemorrh. 1958 May 1;2(1-2):24–38. [PubMed] [Google Scholar]
  2. Bevilacqua M. P., Pober J. S., Majeau G. R., Cotran R. S., Gimbrone M. A., Jr Interleukin 1 (IL-1) induces biosynthesis and cell surface expression of procoagulant activity in human vascular endothelial cells. J Exp Med. 1984 Aug 1;160(2):618–623. doi: 10.1084/jem.160.2.618. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Bradford M. M. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem. 1976 May 7;72:248–254. doi: 10.1006/abio.1976.9999. [DOI] [PubMed] [Google Scholar]
  4. Canfield W. M., Kisiel W. Evidence of normal functional levels of activated protein C inhibitor in combined Factor V/VIII deficiency disease. J Clin Invest. 1982 Dec;70(6):1260–1272. doi: 10.1172/JCI110725. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Cerveny T. J., Fass D. N., Mann K. G. Synthesis of coagulation factor V by cultured aortic endothelium. Blood. 1984 Jun;63(6):1467–1474. [PubMed] [Google Scholar]
  6. Comp P. C., Esmon C. T. Activated protein C inhibits platelet prothrombin-converting activity. Blood. 1979 Dec;54(6):1272–1281. [PubMed] [Google Scholar]
  7. Dudman N. P., Wilcken D. E. Homocysteine thiolactone and experimental homocysteinemia. Biochem Med. 1982 Apr;27(2):244–253. doi: 10.1016/0006-2944(82)90027-8. [DOI] [PubMed] [Google Scholar]
  8. Esmon C. T., Owen W. G. Identification of an endothelial cell cofactor for thrombin-catalyzed activation of protein C. Proc Natl Acad Sci U S A. 1981 Apr;78(4):2249–2252. doi: 10.1073/pnas.78.4.2249. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Foster W. B., Nesheim M. E., Mann K. G. The factor Xa-catalyzed activation of factor V. J Biol Chem. 1983 Nov 25;258(22):13970–13977. [PubMed] [Google Scholar]
  10. Harker L. A., Slichter S. J., Scott C. R., Ross R. Homocystinemia. Vascular injury and arterial thrombosis. N Engl J Med. 1974 Sep 12;291(11):537–543. doi: 10.1056/NEJM197409122911101. [DOI] [PubMed] [Google Scholar]
  11. Hill-Zobel R. L., Pyeritz R. E., Scheffel U., Malpica O., Engin S., Camargo E. E., Abbott M., Guilarte T. R., Hill J., McIntyre P. A. Kinetics and distribution of 111Indium-labeled platelets in patients with homocystinuria. N Engl J Med. 1982 Sep 23;307(13):781–786. doi: 10.1056/NEJM198209233071303. [DOI] [PubMed] [Google Scholar]
  12. Isaacs J. D., Savion N., Gospodarowicz D., Fenton J. W., 2nd, Shuman M. A. Covalent binding of thrombin to specific sites on corneal endothelial cells. Biochemistry. 1981 Jan 20;20(2):398–403. doi: 10.1021/bi00505a027. [DOI] [PubMed] [Google Scholar]
  13. Jaffe E. A., Nachman R. L., Becker C. G., Minick C. R. Culture of human endothelial cells derived from umbilical veins. Identification by morphologic and immunologic criteria. J Clin Invest. 1973 Nov;52(11):2745–2756. doi: 10.1172/JCI107470. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Kane W. H., Majerus P. W. Purification and characterization of human coagulation factor V. J Biol Chem. 1981 Jan 25;256(2):1002–1007. [PubMed] [Google Scholar]
  15. Kane W. H., Majerus P. W. The interaction of human coagulation factor Va with platelets. J Biol Chem. 1982 Apr 10;257(7):3963–3969. [PubMed] [Google Scholar]
  16. Kane W. H., Mruk J. S., Majerus P. W. Activation of coagulation factor V by a platelet protease. J Clin Invest. 1982 Nov;70(5):1092–1100. doi: 10.1172/JCI110697. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Kettner C., Shaw E. Inactivation of trypsin-like enzymes with peptides of arginine chloromethyl ketone. Methods Enzymol. 1981;80(Pt 100):826–842. doi: 10.1016/s0076-6879(81)80065-1. [DOI] [PubMed] [Google Scholar]
  18. Kisiel W., Canfield W. M., Ericsson L. H., Davie E. W. Anticoagulant properties of bovine plasma protein C following activation by thrombin. Biochemistry. 1977 Dec 27;16(26):5824–5831. doi: 10.1021/bi00645a029. [DOI] [PubMed] [Google Scholar]
  19. LEWIS M. L., WARE A. G. A one-stage method for the determination of accelerator globulin. Proc Soc Exp Biol Med. 1953 Dec;84(3):640–643. doi: 10.3181/00379727-84-20738. [DOI] [PubMed] [Google Scholar]
  20. Loskutoff D. J., Edgington T. E. Synthesis of a fibrinolytic activator and inhibitor by endothelial cells. Proc Natl Acad Sci U S A. 1977 Sep;74(9):3903–3907. doi: 10.1073/pnas.74.9.3903. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Maynard J. R., Dreyer B. E., Stemerman M. B., Pitlick F. A. Tissue-factor coagulant activity of cultured human endothelial and smooth muscle cells and fibroblasts. Blood. 1977 Sep;50(3):387–396. [PubMed] [Google Scholar]
  22. Miletich J. P., Broze G. J., Jr, Majerus P. W. The synthesis of sulfated dextran beads for isolation of human plasma coagulation factors II, IX, and X. Anal Biochem. 1980 Jul 1;105(2):304–310. doi: 10.1016/0003-2697(80)90462-5. [DOI] [PubMed] [Google Scholar]
  23. Nesheim M. E., Taswell J. B., Mann K. G. The contribution of bovine Factor V and Factor Va to the activity of prothrombinase. J Biol Chem. 1979 Nov 10;254(21):10952–10962. [PubMed] [Google Scholar]
  24. RAPAPORT S. I., SCHIFFMAN S., PATCH M. J., AMES S. B. The importance of activation of antihemophilic globulin and proaccelerin by traces of thrombin in the generation of intrinsic prothrombinase activity. Blood. 1963 Feb;21:221–236. [PubMed] [Google Scholar]
  25. Ratnoff O. D. Activation of Hageman factor by L-homocystine. Science. 1968 Nov 29;162(3857):1007–1009. doi: 10.1126/science.162.3857.1007-a. [DOI] [PubMed] [Google Scholar]
  26. Rodgers G. M., Greenberg C. S., Shuman M. A. Characterization of the effects of cultured vascular cells on the activation of blood coagulation. Blood. 1983 Jun;61(6):1155–1162. [PubMed] [Google Scholar]
  27. Rodgers G. M., Shuman M. A. Characterization of the interaction between factor Xa and bovine aortic endothelial cells. Biochim Biophys Acta. 1985 Mar 21;844(3):320–329. doi: 10.1016/0167-4889(85)90133-8. [DOI] [PubMed] [Google Scholar]
  28. Rodgers G. M., Shuman M. A. Prothrombin is activated on vascular endothelial cells by factor Xa and calcium. Proc Natl Acad Sci U S A. 1983 Nov;80(22):7001–7005. doi: 10.1073/pnas.80.22.7001. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Rosenberg R. D., Rosenberg J. S. Natural anticoagulant mechanisms. J Clin Invest. 1984 Jul;74(1):1–6. doi: 10.1172/JCI111389. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Sinha A. K., Dutta-Roy A. K., Chiu H. C., Stewart G. J., Colman R. W. Coagulant factor Xa inhibits prostacyclin formation in human endothelial cells. Role of factor V. Arteriosclerosis. 1985 May-Jun;5(3):244–249. doi: 10.1161/01.atv.5.3.244. [DOI] [PubMed] [Google Scholar]
  31. Stern D. M., Drillings M., Nossel H. L., Hurlet-Jensen A., LaGamma K. S., Owen J. Binding of factors IX and IXa to cultured vascular endothelial cells. Proc Natl Acad Sci U S A. 1983 Jul;80(13):4119–4123. doi: 10.1073/pnas.80.13.4119. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Stern D. M., Nawroth P. P., Harris K., Esmon C. T. Cultured bovine aortic endothelial cells promote activated protein C-protein S-mediated inactivation of factor Va. J Biol Chem. 1986 Jan 15;261(2):713–718. [PubMed] [Google Scholar]
  33. Stern D. M., Nawroth P. P., Kisiel W., Handley D., Drillings M., Bartos J. A coagulation pathway on bovine aortic segments leading to generation of Factor Xa and thrombin. J Clin Invest. 1984 Dec;74(6):1910–1921. doi: 10.1172/JCI111611. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Tracy P. B., Nesheim M. E., Mann K. G. Proteolytic alterations of factor Va bound to platelets. J Biol Chem. 1983 Jan 10;258(1):662–669. [PubMed] [Google Scholar]
  35. Tracy P. B., Peterson J. M., Nesheim M. E., McDuffie F. C., Mann K. G. Interaction of coagulation factor V and factor Va with platelets. J Biol Chem. 1979 Oct 25;254(20):10354–10361. [PubMed] [Google Scholar]
  36. Uhlemann E. R., TenPas J. H., Lucky A. W., Schulman J. D., Mudd S. H., Shulman N. R. Platelet survival and morphology in homocystinuria due to cystathionine synthase deficiency. N Engl J Med. 1976 Dec 2;295(23):1283–1286. doi: 10.1056/NEJM197612022952303. [DOI] [PubMed] [Google Scholar]
  37. Wall R. T., Harlan J. M., Harker L. A., Striker G. E. Homocysteine-induced endothelial cell injury in vitro: a model for the study of vascular injury. Thromb Res. 1980 Apr 1;18(1-2):113–121. doi: 10.1016/0049-3848(80)90175-9. [DOI] [PubMed] [Google Scholar]
  38. Weksler B. B., Marcus A. J., Jaffe E. A. Synthesis of prostaglandin I2 (prostacyclin) by cultured human and bovine endothelial cells. Proc Natl Acad Sci U S A. 1977 Sep;74(9):3922–3926. doi: 10.1073/pnas.74.9.3922. [DOI] [PMC free article] [PubMed] [Google Scholar]
  39. Wiggins R. C., Loskutoff D. J., Cochrane C. G., Griffin J. H., Edgington T. S. Activation of rabbit Hageman factor by homogenates of cultured rabbit endothelial cells. J Clin Invest. 1980 Jan;65(1):197–206. doi: 10.1172/JCI109651. [DOI] [PMC free article] [PubMed] [Google Scholar]
  40. Wight T. N. Vessel proteoglycans and thrombogenesis. Prog Hemost Thromb. 1980;5:1–39. [PubMed] [Google Scholar]
  41. de Groot P. G., Willems C., Boers G. H., Gonsalves M. D., van Aken W. G., van Mourik J. A. Endothelial cell dysfunction in homocystinuria. Eur J Clin Invest. 1983 Oct;13(5):405–410. doi: 10.1111/j.1365-2362.1983.tb00121.x. [DOI] [PubMed] [Google Scholar]

Articles from Journal of Clinical Investigation are provided here courtesy of American Society for Clinical Investigation

RESOURCES