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Thromb Haemost 1992; 67(06): 654-659
DOI: 10.1055/s-0038-1648518
Original Articles
Schattauer GmbH Stuttgart

Factor Vlla/Tissue Factor-Catalyzed Activation of Factors IX and X on a Cell Surface and in Suspension: A Kinetic Study

L Vijaya Mohan Rao
The Department of Medicine, University of California, San Diego, CA, USA
,
Thomas Robinson
The Department of Medicine, University of California, San Diego, CA, USA
,
An D Hoang
The Department of Medicine, University of California, San Diego, CA, USA
› Author Affiliations
Further Information

Publication History

Received 28 August 1991

Accepted after revision 20 January 1992

Publication Date:
03 July 2018 (online)

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Summary

The kinetics of activation of factor IX and factor X by factor Vila was studied in the presence of various sources of tissue factor: (1) a surface membrane of human ovarian carcinoma cell line, OC-2008 (2) the cell lysate (of 002008) and (3) reconstituted purified human tissue factor. The rates of activation of factors IX and X were monitored in activation peptide release assays using tritiated substrates. The results indicate that the apparent K m values for factor IX and factor X were similar for a given tissue factor, but varied with tissue factor source. The source of tissue factor greatly influenced the apparent differences in V max for factors IX and X. When a surface of monolayer provided tissue factor, the Vmax of factor IX was only 2-3 fold lower than factor X, but when either a cell lysate or purified tissue factor was the source of cofactor activity, the difference in V max rose to about 8-10 fold. Although, the tissue factor apoprotein in the cells was expressed entirely on the outer surface membrane, the activity of tissue factor on the intact cell surface was 50 to 100fold lower than in the lysed cell preparation.

 

  • References

  • 1 Nemerson Y. Tissue factor and hemostasis. Blood 1988; 71: 1-8
    PubMedGoogle Scholar
  • 2 Silverberg SA, Nemerson Y, Zur M. Kinetics of the activation of bovine coagulation factor X by components of the extrinsic pathway. Kinetic behavior of two-chain factor VII in the presence and absence of tissue factor. J Biol Chem 1977; 252: 8481-8488
    PubMedGoogle Scholar
  • 3 Jesty J, Silverberg SA. Kinetics of the tissue factor-dependent activation of coagulation factors IX and X in a bovine plasma system. J Biol Chem 1979; 254: 1233-1245
    PubMedGoogle Scholar
  • 4 Zur M, Nemerson Y. Kinetics of factor IX activation via the extrinsic pathway. Dependence of Km on tissue factor. J Biol Chem 1980; 255: 5703-5707
    PubMedGoogle Scholar
  • 5 Jesty J, Morrison SA. The activation of factor IX by tissue factor-factor VII in a bovine plasma lacking factor X. Thromb Res 1983; 32: 171-181
    CrossrefPubMedGoogle Scholar
  • 6 Morrison SA, Jesty J. Tissue factor-dependent activation of tritium labeled factor IX and factor X in human plasma. Blood 1983; 63: 1338-1347
    PubMedGoogle Scholar
  • 7 Warn-Cramer BJ, Bajaj SP. Intrinsic versus extrinsic coagulations. Kinetic consideration. Biochem J 1986; 239: 757-762
    CrossrefPubMedGoogle Scholar
  • 8 Bom VJJ, Reinalda-Poot JH, Cupers R, Bertina RM. Extrinsic activation of human blood coagulation factors IX and X. Thromb Haemostas 1990; 63: 224-230
    PubMedGoogle Scholar
  • 9 Komiyama Y, Pedersen A, Kisiel W. Proteolytic activation of human factors IX and X by recombinant human factor Vila. Effects of calcium, phospholipids, and tissue factor. Biochemistry 1990; 29: 9418-9425
    CrossrefPubMedGoogle Scholar
  • 10 Bom VJJ, van Hinsbergh VWM, Reinalda-Poot HH, Mohanlal RW, Bertina RM. Extrinsic activation of human coagulation factors IX and X on the endothelial surface. Thromb Haemostas 1991; 66: 283-291
    PubMedGoogle Scholar
  • 11 Almus FE, Rao LVM, Rapaport SI. Functional properties of factor Vlla/tissue factor formed with purified tissue factor and with tissue factor expressed on cultured endothelial cells. Thromb Haemostas 1989; 62: 1067-1072
    PubMedGoogle Scholar
  • 12 Almus FE, Rao LVM, Rapaport SI. Properties of factor Vlla/tissue factor in an umbilical vein model. Blood 1990; 76: 354-360
    PubMedGoogle Scholar
  • 13 Rao LVM, Hoang AD. Expression of tissue factor in human ovarian carcinoma cell line. Blood 1989; 74: 96 (abstr)
    PubMedGoogle Scholar
  • 14 Rickies FR, Edwards RL. Activation of blood coagulation in cancer: Trousseau’s syndrome revisited. Blood 1983; 62: 14-31
    PubMedGoogle Scholar
  • 15 Rao LVM, Bajaj SP. Purification of human factor VII utilizing O-(Diethylaminoethyl)-Sephadex and Sulfopropyl Sephadex chromatography. Anal Biochem 1984; 136: 357-361
    CrossrefPubMedGoogle Scholar
  • 16 Bajaj SP, Rapaport SI, Prodanos C. A simplified procedure for purification of human prothrombin, factor IX and factor X. Prep Biochem 1981; 11: 397-412
    PubMedGoogle Scholar
  • 17 Rao LVM. Characterization of anti-human tissue factor antibody and its use in immunoaffinity purification of human tissue factor. Thromb Res 1988; 51: 373-384
    CrossrefPubMedGoogle Scholar
  • 18 Rao LVM, Rapaport SI. Activation of factor VII bound to tissue factor: A key early step in the tissue factor pathway of blood coagulation. Proc Natl Acad Sci USA 1988; 85: 6687-6691
    CrossrefPubMedGoogle Scholar
  • 19 Mimms LT, Zamphighi G, Nozaki Y, Tanford C, Reynolds J. Phospholipid vesicle formation and transmembrane protein incorporation. Biochemistry 1981; 20: 840-849
    CrossrefPubMedGoogle Scholar
  • 20 Van Lenten L, Ashwell G. Studies on the chemical and enzymatic modification of glycoproteins. J Biol Chem 1979; 246: 1889-1894
    PubMedGoogle Scholar
  • 21 Sanders NL, Bajaj SP, Zivelin A, Rapaport SI. Inhibition of tissue factor/factor Vila activity in plasma requires factor X and an additional plasma component. Blood 1985; 66: 204-212
    PubMedGoogle Scholar
  • 22 Laemmli UK. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 1970; 227: 680-685
    CrossrefPubMedGoogle Scholar
  • 23 McGee MP, Wallin R, Wheeler FB, Rothberger H. Initiation of the extrinsic pathway of coagulation by human and rabbit alveolar macrophages: A kinetic studv. Blood 1989; 74: 1583-1590
    PubMedGoogle Scholar
  • 24 Bach R, Rifkin DB. Expression of tissue factor procoagulant activity: Regulation by cytosolic calcium. Proc Natl Acad Sci USA 1990; 87: 6995-6999
    CrossrefPubMedGoogle Scholar
  • 25 Le DT, Rapaport SI, Rao LVM. Relations between F Vila binding and tissue factor activity of an ovarian carcinoma cell line (OC-2008). Blood 1991; 78: 238 (abstr)
    PubMedGoogle Scholar
  • 26 Drake TA, Ruf W, Morrissey JH, Edgington TS. Functional tissue factor is entirely cell surface expressed on lipopolysaccharide-stimu-lated human blood monocytes and a constitutively tissue factor producing neoplastic cell line. J Cell Biol 1989; 109: 389-394
    CrossrefPubMedGoogle Scholar
  • 27 Op den Kamp JAF. Lipid asymmetry in membranes. Annu Rev Biochem 1979; 48: 47-71
    CrossrefPubMedGoogle Scholar
  • 28 Ryan J, Brett J, Kisiel W, Stern D. Tumor necrosis (TNF)-induced endothelial cell tissue factor is principally associated with subendothelial matrix. Blood 1990; 76: 2068 (abstr)
    PubMedGoogle Scholar