Advertisement
Paper| Volume 40, ISSUE 6, P745-756, December 15, 1985

Purification of glycosylated apoprotein of tissue factor from human brain and inhibition of its procoagulant activity by a specific antibody

  • H. Tanaka
    Affiliations
    Clinical Research Unit for Blood Coagulation and Thrombosis of the Max-Planck-Gesellschaft at the Justus-Liebig-Universitat Gaffkystr. 11, D-6300 Giessen, West-Germany
    Search for articles by this author
  • B. Janssen
    Affiliations
    Clinical Research Unit for Blood Coagulation and Thrombosis of the Max-Planck-Gesellschaft at the Justus-Liebig-Universitat Gaffkystr. 11, D-6300 Giessen, West-Germany
    Search for articles by this author
  • K.T. Preissner
    Affiliations
    Clinical Research Unit for Blood Coagulation and Thrombosis of the Max-Planck-Gesellschaft at the Justus-Liebig-Universitat Gaffkystr. 11, D-6300 Giessen, West-Germany
    Search for articles by this author
  • G. Müller-Berghaus
    Correspondence
    To whom correspondence should be addressed
    Affiliations
    Clinical Research Unit for Blood Coagulation and Thrombosis of the Max-Planck-Gesellschaft at the Justus-Liebig-Universitat Gaffkystr. 11, D-6300 Giessen, West-Germany
    Search for articles by this author
DOI:https://doi.org/10.1016/0049-3848(85)90312-3
Purification of glycosylated apoprotein of tissue factor from human brain and inhibition of its procoagulant activity by a specific antibody
Previous ArticleThrombus formation by the application of thrombin to the outer surface of mouse mesenteric vein: Comparison with the application of ADP
Next ArticleCharacterization of human platelet beta-adrenoceptors
      This paper is only available as a PDF. To read, Please Download here.

      Abstract

      The glycosylated component of tissue factor from human brain which totally retained on Concanavalin A-Sepharose was isolated by a four step procedure. The final product (6–7 S) exhibited an apparent molecular weight of approximately 45, 000 under reducing conditions on SDS-gels corresponding with procoagulant activity of tissue factor after detergent removal. The optimal ratio of lipid to protein in reconstituting tissue factor activity was 500:1 (w/w) amounting to 240, 000 U/mg. Peptidase activity was not detectable in the purified apoprotein or in the relipidated protein. The procoagulant activity of tissue factor was dependent on Factor VII but not on Factors VIII or IX, since in the absence of Factor VII clotting times were significantly prolonged. The activity of the reconstituted tissue factor could be reduced by a factor of 15 upon incubation with an antibody against the purified glycosylated apoprotein that was raised in the rabbit. The IgG-fraction of the antiserum neutralized the procoagulant activity of tissue factor in saline extracts from human brain and placenta in a time- and concentration-dependent manner indicating common antigenic determinants on the cofactor protein from both tissues.

      Keywords

      To read this article in full you will need to make a payment

      Purchase one-time access:

      Academic & Personal: 24 hour online accessCorporate R&D Professionals: 24 hour online access
      One-time access price info
      • For academic or personal research use, select 'Academic and Personal'
      • For corporate R&D use, select 'Corporate R&D Professionals'

      Subscribe:

      Subscribe to Thrombosis Research
      Already a print subscriber? Claim online access
      Already an online subscriber? Sign in
      Register: Create an account
      Institutional Access: Sign in to ScienceDirect

      References

        • Nemerson Y.
        • Bach R.
        Tissue factor revisited.
        in: Spaet T.H. Progress in Hemostasis and Thrombosis. Grune & Stratton, New York1982: 237-261 (Vol. 6)
        • Pitlick F.A.
        • Nemerson Y.
        Purification and characterization of tissue factor apoprotein.
        Methods Enzymol. 1976; 45: 37-48
        • Nemerson Y.
        The reaction between bovine brain tissue factor and factor VII and X.
        Biochemistry. 1966; 5: 601-608
        • Zur M.
        • Nemerson Y.
        Kinetics of factor IX activation via the extrinsic pathway.
        in: Dependence of Km on tissue factor. 5th edition. J. Biol. Chem.255. 1980: 5703-5707
        • Van Den Besselaar A.M.H.P.
        • Ram I.E.
        • Alderkamp G.H.J.
        • Bertina R.M.
        The role of factor IX in tissue thromboplastin induced coagulation.
        Thromb. Haemostas. 1982; 48: 54-58
        • Marlar R.A.
        • Kleiss A.J.
        • Griffin J.H.
        An alternative extrinsic pathway of human blood coagulation.
        Blood. 1982; 60: 1353-1358
        • Morrison S.A.
        • Jesty J.
        Tissue factor-dependent activation of tritiumlabeled factor IX and X in human plasma.
        Blood. 1984; 63: 1338-1347
        • Glas P.
        • Astrup T.
        Thromboplastin and plasminogen activator in tissues of the rabbit.
        Am. J. Physiol. 1970; 219: 1140-1146
        • Rivers R.P.A.
        • Hathaway W.E.
        • Weston W.L.
        The endotoxin induced coagulant activity of human monocytes.
        Br. J. Haematol. 1975; 30: 311-316
        • Prydz H.
        • Allison A.C.
        Tissue thromboplastin activity of isolated human monocytes.
        Thromb. Haemostas. 1978; 39: 582-591
        • Prydz H.
        • Lyberg T.
        • Deteix P.
        • Allison A.C.
        In vitro stimulation of tissue thromboplastin (Factor III) activity in human monocytes by immune complexes and lectins.
        Thromb. Res. 1979; 15: 465-474
        • Edwards R.L.
        • Rickles F.R.
        The role of human T cells (and T cell products) for monocyte tissue factor generation.
        J. Immunol. 1980; 125: 606-609
        • Prydz H.
        • Allison A.C.
        • Schorlemmer H.U.
        Further link between complement activation and blood coagulation.
        Nature. 1977; 270: 173-174
        • Lyberg T.
        • Galdal K.S.
        • Evensen S.A.
        • Prydz H.
        Cellular cooperation in endothelial cell thromboplastin synthesis.
        Br. J. Haematol. 1983; 53: 85-95
        • Colucci M.
        • Balconi G.
        • Lorenzet R.
        • Pietra A.
        • Locati D.
        • Donati M.B.
        • Semeraro N.
        Cultured human endothelial cells generate tissue factor in response to endotoxin.
        J. Clin. Invest. 1983; 71: 1893-1896
        • Hvatum M.
        • Prydz H.
        Studies on tissue thromboplastin - its splitting into two separable parts.
        Thromb. Diath. Haemorrh. 1969; 21: 217-222
        • Bjorklid E.
        • Storm E.
        • Prydz H.
        The protein component of human brain thromboplastin.
        Biochem. Biophys. Res. Commun. 1973; 55: 969-976
        • Bjorklid E.
        • Storm E.
        Purification and some properties of the protein component of tissue thromboplastin from human brain.
        Biochem. J. 1977; 165: 89-96
        • Gonmori H.
        • Takeda Y.
        Properties of human tissue thromboplastins from brain, lung, arteries and placenta.
        Thromb. Haemostas. 1976; 36: 90-103
        • Carson S.D.
        • Konigsberg W.H.
        Lipid activation of coagulation factor III apoprotein (tissue factor) - reconstitution of the protein-membrane complex.
        Thromb. Haemostas. 1980; 44: 12-15
        • Kurosawa S.
        • Matsuda M.
        • Aoki N.
        Urinary procoagulant behaves as tissue factor by promoting factor VIIa-catalyzed activation of factor X.
        Thromb. Res. 1984; 33: 595-606
        • Nemerson Y.
        • Pitlick F.A.
        Purification and characterization of the protein component of tissue factor.
        Biochemistry. 1970; 9: 5100-5105
        • Lui D.T.H.
        • McCoy L.E.
        Tissue extract thromboplastin: Quantitation, fractionation and characterization of protein components.
        Thromb. Res. 1975; 7: 199-211
        • Bach R.
        • Nemerson Y.
        • Konigsberg W.
        Purification and characterization of bovine tissue factor.
        J. Biol. Chem. 1981; 256: 8324-8331
        • Gonmori H.
        • Takeda Y.
        Properties of canine tissue thromboplastins of brain, lung, arteries, and veins.
        Am. J. Physiol. 1975; 229: 618-626
        • Howell R.M.
        • Rezvan H.
        Circular dichroic, infrared and other studies on the protein component of pig brain thromboplastin.
        Biochem. J. 1980; 189: 209-218
        • Gardiner J.E.
        • Howell R.M.
        Partial purification of a native form of thromboplastin from procine brain.
        Biochem. Soc. Trans. 1980; 8: 133
        • Tanaka H.
        • Pogatzki G.
        • Preissner K.T.
        • Muller-Berghaus G.
        Purification of rabbit tissue thromboplastin.
        Blut. 1984; 49: 144-145
        • Pitlick F.A.
        Concanavalin A inhibits tissue factor coagulation activity.
        J. Clin. Invest. 1975; 55: 175-179
        • Pitlick F.A.
        Binding of bovine brain tissue factor to concanavalin A Sepharose.
        in: Partial purification of coagulant, arylamidase, and alkaline phosphatase activities. 5th edition. Biochim. Biophys. Acta. 428. 1976: 27-34
        • Laemmli U.K.
        Cleavage of structural proteins during the assembly of the head of bacteriophage T4.
        Nature. 1970; 227: 680-685
        • 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; 72: 248-254
        • Rouser G.
        • Siakotos A.N.
        • Fleischer S.
        Quantitative analysis of phospholipids by thin-layer chromatography and phosphorus analysis of spots.
        Lipids. 1966; 1: 85-86
        • Fiske C.H.
        • Subbarow Y.
        The colorimetric determination of phosphorus.
        J. Biol. Chem. 1925; 66: 375-400
        • Brunner J.
        • Skrabal P.
        • Hauser H.
        Single bilayer vesicles prepared without sonication.
        in: Physico-chemical properties. 5th edition. Biochim. Biophys. Acta. 455. 1976: 322-331
        • Nemerson Y.
        The phospholipid requirement of tissue factor in blood coagulation.
        J. Clin. Invest. 1968; 47: 72-79
        • Nemerson Y.
        Characteristics and lipid requirements of coagulant proteins extracted from lung and brain: The specificity of the protein component of tissue factor.
        J. Clin. Invest. 1969; 48: 322-331
        • O'Brien R.D.
        • Timpone C.A.
        • Gibson R.E.
        The measurement of partial specific volumes and sedimentation coefficients by sucrose density centrifugation.
        Anal. Biochem. 1978; 86: 602-607
        • Ganrot P.O.
        Crossed immunoelectrophoresis.
        Scand. J. Clin. Lab. Invest. 1972; 29: 39-47
        • Van den Besselaar A.M.H.P.
        • Bertina R.M.
        Interaction of thromboplastin apoprotein of different tissues with concanavalin A.
        in: Evidence for heterogeneous glycosylation of the human apoprotein. 5th edition. Thromb. Haemostas.52. 1984: 192-195

      Article info

      Publication history

      Accepted: September 5, 1985
      Received: May 29, 1985

      Identification

      DOI: https://doi.org/10.1016/0049-3848(85)90312-3

      Copyright

      © 1985 Published by Elsevier Inc.

      ScienceDirect

      Access this article on ScienceDirect

      The content on this site is intended for healthcare professionals.


      We use cookies to help provide and enhance our service and tailor content. To update your cookie settings, please visit the Cookie Preference Center for this site.
      Copyright © 2023 Elsevier Inc. except certain content provided by third parties.


      RELX