Structural and Functional Properties of Protein C

  • Frederick J. Walker


The discovery of protein C and its effects upon blood coagulation, fibrinolysis, and inflammation has changed the way we now think about the pathophysiology of thromboembolic disease and related disorders. Interestingly, this discovery was not the result of a planned approach to understanding a disease, but rather from a basic approach of attempting to understand the biochemistry of the vitamin K-dependent family of proteins. In fact all of the clinical information about protein C stems from information obtained in basic research laboratories. The first clue of the existence of protein C can be found in a 1947 paper that reports that factor Va is unstable in serum (1). It was subsequently observed that the instability could be removed if the plasma was adsorbed with aluminum hydroxide, an insoluble support that was known to adsorb vitamin K-dependent proteins. Seegers group observed that treatment of prothrombin with thrombin could elicit an inhibitor of coagulation. This led to the inhibitor’s characterization and designation as autoprothrombin IIa (2). Marciniak (3) confirmed the observation and noted that the inhibitor was species specific. Some time later, Stenflo characterized a new vitamin K-dependent protein (4), named protein C (5) which turned out to be the zymogen of autoprothrombin IIa (6). Characterization of protein C and the discovery that activated protein C was a potent inhibitor of blood coagulation lead to the realization that protein C may be one of the keys to understanding the molecular basis of thrombotic disease. This paper describes structural properties that give rise both to its unique functional — properties as well as the complex scheme by which it is regulated.


Factor VIII Cofactor Activity Epidermal Growth Factor Domain Euglobulin Clot Lysis Time Blood Clot Lysis 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.


Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.


  1. 1.
    Owren PA: The coagulation of blood. Acta Med. Scand. Suppl. 194: 6–56, 1947Google Scholar
  2. 2.
    Mammen EF, Thomas WR, Seegers WH: Activation of purified prothrombin to autoprothrombin I or autoprothrombin II (platelet cofactor II) or autoprothrombin IIa. Thrombos. Diath. Haemorrh. 5: 218–249, 1960Google Scholar
  3. 3.
    Marcinak E.: Coagulation inhibitor elicited by thrombin. Science 170: 452–453, 1970CrossRefGoogle Scholar
  4. 4.
    Stenflo J: A new vitamin K-dependent protein. J. Biol. Chem. 251: 255–263, 1976Google Scholar
  5. 5.
    Esmon CT, Stenflo J, Suttie JW, Jackson CM: A new vitamin K-dependent protein A phospholipid-binding zymogen of a serine protease. J. Biol. Chem. 251: 3052–3056, 1976PubMedGoogle Scholar
  6. 6.
    Seegers WH, Novoa E, Henry RL, Hassouna HI: Relationship of “new” vitamin K-dependent protein C and “old” autoprothrombin IIA. Thromb. Res 8: 543–546, 1976PubMedCrossRefGoogle Scholar
  7. 7.
    Kisiel W, Ericsson LH, Davie EW: Proteolytic activation of protein C from bovine plasma. Biochemistry 15: 4893–4900, 1976PubMedCrossRefGoogle Scholar
  8. 8.
    Fair DS, Marlar RA: Biosynthesis and secretion of factor VII, protein C, protein S, and the protein C inhibitor from a human hepatoma cell line. Blood 67: 64–70, 1986PubMedGoogle Scholar
  9. 9.
    Fernlund P, Stenflo J: Amino acid sequence of the light chain of bovine protein C. J. Biol. Chem. 257: 12170–12179, 1982PubMedGoogle Scholar
  10. 10.
    Fernlund P, Stenflo, J: Amino acid sequence of the heavy chain of bovine protein C. J. Biol. Chem.. 257: 12180–12190, 1982PubMedGoogle Scholar
  11. 11.
    Ohlin A, Landes G, Bourdon P, Oppenheimer C, Wydro R, Stenflo J: beta hydroxyaspartic acid in the first epidermal growth factor-like domain of protein C. Its role in calcium binding and biological activity. J. Biol. Chem. 263: 19240–19248, 1988PubMedGoogle Scholar
  12. 12.
    Amphlett GF, Kisiel W, Castellino, FJ: Interaction of calcium with bovine plasma protein C. Biochemistry 20: 2156–2162, 1981PubMedCrossRefGoogle Scholar
  13. 13.
    Esmon N, DeBault L, Esmon C: Proteolytic formation and properties of gamma-carboxyglutamic acid domainless protein C. J. Biol. Chem. 258: 5548–5553, 1983PubMedGoogle Scholar
  14. 14.
    Johnson AE, Esmon N, Laue T, Esmon CT: Structural changes required for activation of protein C are induced by calcium binding to a high affinity site that does not contain gamma carboxyglutamic acid. J. Biol. Chem. 258: 5554–5560, 1983PubMedGoogle Scholar
  15. 15.
    Ohlin A, Linse S, Stenflo J: Calcium binding to the epidermal growth factor homology region of bovine protein C. J. Biol. Chem. 263: 7411–7417, 1988PubMedGoogle Scholar
  16. 16.
    Church WR, Bhushan FH, Mann KG, Bovill EG: Discrimination of normal and abnormal prothrombin and protein C in plasma using calcium ion inhibited monoclonal antibody to a common epitope on several vitamin K-dependent proteins. Blood 74: 2418–2425, 1989PubMedGoogle Scholar
  17. 17.
    Church WR, Messier T, Horare PJ, Amiral, D. Meyer, Mann KG: A conserved epitope on several human vitamin k-dependent proteins. J. Biol. Chem. 263: 6259–6267, 1988PubMedGoogle Scholar
  18. 18.
    Orthner CL, Madurawe RD, Velander WH, Drohan WN, Battey FD, Strickland DK: Conformational changes in an epitope localized in the NH2-terminal region of protein C. Evidence for interaction of protein C domains. J. Biol. Chem. 264: 18781–18788, 1989PubMedGoogle Scholar
  19. 19.
    Hill KA, Castellino FJ: The binding of manganese to bovine plasma protein C des(1-41)-light chain protein C and activated des (1-41)-light chain activated protein C. Arch. Biochem. Biophys. 254: 196–202, 1987PubMedCrossRefGoogle Scholar
  20. 20.
    Steiner SA, Castellino FJ: Kinetic Mechanism for stimulation by monovalent cations of the amidase activity of the plasma protein bovine activated protein C. Biochemistry 24: 609–617, 1985PubMedCrossRefGoogle Scholar
  21. 21.
    Steiner SA, Castellino FJ: Kinetic studies of the role of monovalent cations in the amidolytic activity of activated bovine plasma protein C. Biochemistry 21: 4609–4614, 1982PubMedCrossRefGoogle Scholar
  22. 22.
    Hill KAW, Castellino FJ: The stimulation by monovalent cations of the amidase activity of bovine des 1-41 light chain activated protein C. J. Biol. Chem. 261: 14991–14996, 1986PubMedGoogle Scholar
  23. 23.
    Hill KAW, Castellino FJ: The effect of monovalent cations on the presteady state reaction kinetics of bovine activated protein C and des 1-41 light chain activated protein C. J. Biol. Chem. 262: 140–146, 1987PubMedGoogle Scholar
  24. 24.
    Hill KAW, Castellino FJ, Tl3+ as a spectroscopic probe of the monovalent cation binding site of bovine plasma activated protein C and des 1-41 light chain activated protein C. J. Biol. Chem. 262: 7098–7104, 1987PubMedGoogle Scholar
  25. 25.
    Hill KAW, Castellino FJ: Topographical relationships amoung the monovalent cation binding sites of bovine plasma activated protein C and des(1-41) light chain activated protein C and a nitroxide spin label bound to their active site serine residues. J. Protein Chemistry 6: 489–496, 1987Google Scholar
  26. 26.
    Haley PE, Doyle MF, Mann KG: The activation of bovine protein C by factor Xa. J. Biol. Chem. 264: 16303–16310, 1989PubMedGoogle Scholar
  27. 27.
    Walker FJ, Sexton PW, Esmon CT: The inhibition of blood coagulation by activated protein C through the selective inactivation of activated factor V. Biochim. Biophys. Acta 571: 333–342, 1979PubMedCrossRefGoogle Scholar
  28. 28.
    Klein JD, Walker FJ: Purification of a protein C activator from the venom of the southern coopperhead snake (Agkistrodon Contortrix Contortrix). Biochemistry 25: 4175–4179, 1986PubMedCrossRefGoogle Scholar
  29. 29.
    Orthner CL, Bhattacharya P, Strickland DK: Characterization of a protein C activator from the venom of Agkistrodon contortrix contortrix. Biochemistry 27: 2558–2564, 1988PubMedCrossRefGoogle Scholar
  30. 30.
    Francis RB, Seyfert U: Rapid amidolytic assay of protein C in whole plasma using an activator from the venom of Agkistrodon Contortrix. A.J.C.P. 87: 619–625, 1987Google Scholar
  31. 31.
    Esmon CT, Owen WG: Identification of an endothelial cell cofactor for thrombin-catalyzed activation of protein C. Proc. Natl. Acad. Sci. U.S.A. 78: 2249–2252, 1981PubMedCrossRefGoogle Scholar
  32. 32.
    Esmon NL, Owen WG, Esmon CT: Isolation of membrane-bound cofactor for thrombin-catalyzed activation of protein C. J. Biol. Chem. 257: 859–864, 1982PubMedGoogle Scholar
  33. 33.
    Owen WG, Esmon CT: Functional properties of an endothelial cell cofactor for thrombin-catalyzed activation of protein C. J. Biol. Chem. 256: 5532–5537, 1981PubMedGoogle Scholar
  34. 34.
    Esmon CT, Esmon NL, Harris KW: Complex formation between thrombin and thrombomodulin inhibits both thrombin-catalyzed fibrin formation and factor V activation. J. Biol. Chem. 257: 7944–7950, 1982PubMedGoogle Scholar
  35. 35.
    Esmon NL, Carrol RC, Esmon CT: Thrombomodulin blocks the ability of thrombin to activate platelets. J. Biol. Chem. 258: 12238–12242, 1983PubMedGoogle Scholar
  36. 36.
    Bourin MC, Ohlin AK, Lane DA, Stenflo J, Lindahl U: Relationship between anticoagulant activities and polyanionic properties of rabbit thrombomodulin. J. Biol. Chem. 263: 8044–8052, 1988PubMedGoogle Scholar
  37. 37.
    Hofsteenge J, Taguchi H, Stone SR: Effect of thrombomodulin on the kinetics of the interaction of thrombin with substrates and inhibitors. Biochem J. 237: 243–251, 1986PubMedGoogle Scholar
  38. 38.
    Preissner KT, Delvos U, Muller-Berghaus G: Binding of thrombin to thrombomodulin accelerates inhibition of the enzyme by antithrombin III. Evidence for a heparin-independent mechanism. Biochemistry 26: 2521–2528, 1987PubMedCrossRefGoogle Scholar
  39. 39.
    Dittman WA, Kumada T, Sadler JE, Majerus PW: The structure and function of mouse thrombomodulin. Phorbol myristate acetate stimulates degradation and synthesis of thrombomodulin without affecting mRNA levels in hemangioma cells. J. Biol. Chem. 263: 15815–15822, 1988PubMedGoogle Scholar
  40. 40.
    Suzuki K, Kusumoto H, Hashimoto S: Isolation and characterization of thrombomodulin from bovine lung. Biochim Biophys Acta 882: 343–352, 1986PubMedCrossRefGoogle Scholar
  41. 41.
    Salem HH, Maruyama I, Ishii H, Majerus PW: Isolation and characterization of thrombomodulin from human placenta. J. Biol. Chem. 259: 12246–12251, 1984PubMedGoogle Scholar
  42. 42.
    Kukrosawa S, Aoki N: Preparation of thrombomodulin from human placenta. Thromb. Res. 37: 353–364, 1985CrossRefGoogle Scholar
  43. 43.
    Winnard PT, Esmon CT, Laue TM: The molecular weight and oligomerization of rabbit thrombomodulin as assessed by sedimentation equilibrium. Arch. Biochem. Biophys. 269: 339–344, 1989PubMedCrossRefGoogle Scholar
  44. 44.
    Jackman RW, Beeler DL, VanDeWater L, Rosenberg RD: Characterization of thrombomodulin cDNA reveals structural similarity to the low density lipoprotein receptor. Proc. Natl. Acad. Sci U.S.A. 83: 8834–8838, 1986PubMedCrossRefGoogle Scholar
  45. 45.
    Jackman RW, Beeler DL, Fritze L, Soff G, Rosenberg RD: Human thrombomodulin gene is intron depleted: Nucleic acid sequences of the cDNA and gene predict protein structure and suggest sites of regulatory control. Proc. Natl. Acad. Sci. U.S.A. 84: 6425–6429, 1987PubMedCrossRefGoogle Scholar
  46. 46.
    Stenflo J, Ohlin AK, Owen WG, Schneider WJ: beta-hydroxyaspartic acid or beta-hydroxyasparagine in bovine low density lipoprotein receptor and in bovine thrombomodulin. J. Biol. Chem. 263: 21–24, 1988PubMedGoogle Scholar
  47. 47.
    Petersen TE: The amino terminal domain of thrombomodulin and pancreatic stone protein are homologous with lectins. FEBS Lett. 231: 51–53, 1988PubMedCrossRefGoogle Scholar
  48. 48.
    Jakubowski HV, Kline MD, Owen WG: The effect of bovine thrombomodulin on the specificity of bovine thrombin. J. Biol. Chem. 261: 3876–3882, 1986PubMedGoogle Scholar
  49. 49.
    Bourin, MC: Effect of rabbit thrombomodulin on thrombin inhibition by antithrombin in the presence of heparin. Thromb. Res. 54: 27–39, 1989PubMedCrossRefGoogle Scholar
  50. 50.
    Hirahara K, Koyama M, Matsuishi T, Kukrata M: The effect of human thrombomodulin on the inactivation of thrombin by human antithrombin III. Thromb. Res. 57: 117–126, 1990PubMedCrossRefGoogle Scholar
  51. 51.
    Maruyama I, Salem HH, Ishii H, Majerus PW: Human thrombomodulin is not an efficient inhibitor of the procoagulant activity of thrombin. J. Clin. Invest. 75: 987–991, 1984CrossRefGoogle Scholar
  52. 52.
    Preissner KT, Koyama T, Muller D, Tschopp J, Muller-Berghaus G: Domain structure of the endothelial cell receptor thrombomodulin as deduced from modulation of its anticoagulant functions. Evidence for a glycosaminoglycan-dependent secondary binding site for thrombin. J. Biol. Chem. 265: 4915–4922, 1990PubMedGoogle Scholar
  53. 53.
    Bourin MC, Ohlin AK, Lane DA, Stenflo J, Lindahl U: Relationship between anticoagulant activities and polyanionic properties of rabbit thrombomodulin. J. Biol. Chem. 263: 8044–8052, 1988PubMedGoogle Scholar
  54. 54.
    Bourin MC, Boffa MC, Bjork I, Lindahl U: Functional Domains of rabbit thrombomodulin. Proc. Natl. Acad. Sci.U.S.A. 83: 5924–5928, 1986.PubMedCrossRefGoogle Scholar
  55. 55.
    Preissner KT, Delvos U, Muller-Berghaus G: Binding of thrombin to thrombomodulin accelerates inhibition of the enzyme by antithrombin III. Evidence for a heparin-independent mechanism. Biochemistry 26: 2521–2528, 1987PubMedCrossRefGoogle Scholar
  56. 56.
    Bourin, MC: Effect of rabbit thrombomodulin on thrombin inhibition by antithrombin in the presence of heparin. Thromb. Res. 54: 27–39, 1989PubMedCrossRefGoogle Scholar
  57. 57.
    Jakubowski HV, Owen WG: Macromolecular specificity determinants on thrombin for fibrinogen and thrombomodulin. J. Biol. Chem. 264: 11117–11121, 1989PubMedGoogle Scholar
  58. 58.
    Misci G, Berliner LJ, Esmon CT: Evidence for multiple conformational changes in the active center of thrombin induced by complex formation with thrombomodulin analysis employing nitroxide spin-labels. Biochemistry 27: 769–773, 1988CrossRefGoogle Scholar
  59. 59.
    Musci G, Berliner LJ: Ligands which effect humam protein C activation by thrombin. J. Biol. Chem. 262: 13889–13891, 1987PubMedGoogle Scholar
  60. 60.
    Hofsteenge J., Stone SR: The effect of thrombomodulin on the cleavage of fibrinogen and fibrinogen fragments by thrombin. Eur. J. Biochem. 168: 49–56, 1987PubMedCrossRefGoogle Scholar
  61. 61.
    Lu RL, Esmon NL, Esmon CT, Johnson AE: The active site of the thrombin-thrombomodulin complex. A fluorescence energy transfer measurement of its distance above the membrane surface. J. Biol. Chem. 264: 12956–12962, 1989PubMedGoogle Scholar
  62. 62.
    Kurosawa S, Galvin JB, Esmon NL Esmon CT: Proteolytic formation and properties of functional domains of thrombomodulin. J. Biol. Chem. 262: 2206–2212, 1987PubMedGoogle Scholar
  63. 63.
    Kurosawa S, Stearns D, Jackson K, Esmon CT: A 10-kDa cyanogen bromide fragment from the epidermal growth factor homology domain of rabbit thrombomodulin contains the primary thrombin binding site. J. Biol. Chem. 263: 5993–5996, 1988PubMedGoogle Scholar
  64. 64.
    Stearns D, Kurosawa S, Esmon CT: Microthrombomodulin, Residues 310-486 from the epidermal growth factor precursor homology domain of thrombomodulin will accelerate protein C activation. J. Biol. Chem. 264: 3352–3356, 1989PubMedGoogle Scholar
  65. 65.
    Suzuki K, Hayashi T, Nishioka J, Kosaka Y, Zushi M, Honda G, Yamanoto S: A domain composed of epidermal growth factor-like structures of human thrombomodulin is essential for thrombin binding and for protein C activation. J. Biol. Chem. 264: 4872–4876, 1989PubMedGoogle Scholar
  66. 66.
    Zushi M, Gomi K, Yamamoto S, Maruyama I, Hayashi T, Suzuki K: The last three consecutive epidermal growth factor-like structures of human thrombomodulin comprise the minimum functional domain for protein C-activating cofactor activity and anticoagulant activity. J. Biol. Chem.. 264: 10351–10353, 1989PubMedGoogle Scholar
  67. 67.
    Galvin JB, Kurosawa S, Moore K, Esmon CT, Esmon NL: Reconstitution of rabbit thrombomodulin into phospholipid vesicles. J. Biol. Chem. 262: 2199–2205, 1987PubMedGoogle Scholar
  68. 68.
    Freyssinet JM, Gauchy J, Cazenave JP: The effect of phospholipids on the activation of protein C by the human thrombin-thrombomodulin complex. Biochem J. 238: 151–157, 1986PubMedGoogle Scholar
  69. 69.
    Salem H, Broze G, Miletich J, Majerus P: Human coagulation factor Va is a cofactor for the activation of protein C. Proc. Natl. Acad Sci USA 80: 1584–1588, 1983PubMedCrossRefGoogle Scholar
  70. 70.
    Maruykama I, Salem HH, Majerus PW: Coagulation factor Va binds to human umbilical vein endothelial cells and accelerates protein C activation. J. Clin. Invest. 74: 224–230, 1984CrossRefGoogle Scholar
  71. 71.
    Salem HH, Broze GJ, Miletich JP, Majerus PW: The light chain of factor Va contains the activity of factor Va that accelerates protein C activation by thrombin. J. Biol. Chem. 258: 8531–8534, 1983PubMedGoogle Scholar
  72. 72.
    Salem HH, Esmon NL, Esmon CT, Majerus PW: Effects of thrombomodulin and coagulation factor Va-light chain on protein C activation in vitro. J. Clin Invest. 73: 968–972, 1984PubMedCrossRefGoogle Scholar
  73. 73.
    DeBault LE, Esmon NL, Smith GP, Esmon CT: Localization of thrombomodulin antigen in rabbit endothelial cells in culture. An immunofluorescence and immunoelectron microscope study. Lab. Invest. 54: 179–187, 1986PubMedGoogle Scholar
  74. 74.
    DeBault LE, Esmon NL, Olson JR, Esmon CT: Distribution of the thrombomodulin antigen in the rabbit vasculature. Lab Invest. 54: 172–178, 1986PubMedGoogle Scholar
  75. 75.
    Maruyama I, Bell CE, Majerus PW: Thrombomodulin is found on endothelium of arteries, veins capillaries and lymphatics, and on syncytiotrophoblast of human placenta. J. Cell. Biol. 101: 363–371, 1985PubMedCrossRefGoogle Scholar
  76. 76.
    Ishii H, Salem HH, Bell CE, Laposata EA, Majerus PW: Thrombomodulin, an endothelial anticoagulant protein, is absent from the human brain. Blood 67: 362–365, 1986PubMedGoogle Scholar
  77. 77.
    Freyssinet JM, Gauchy J, Spaethe R, Cazenave JP: Thrombomodulin activity is found in tissue thromboplastin preparations from placenta and from lung but not from brain. Haemostasis, 17: 114–120, 1987PubMedGoogle Scholar
  78. 78.
    Ishii H, Majerus PW: Thrombomodulin is present in human plasma and urine. J. Clin Invest. 76: 2178–2181, 1985PubMedCrossRefGoogle Scholar
  79. 79.
    Beretz A, Freyssinet JM, Gauchy J, Schmitt DA, Klein-Soyer C, Edgell CJ, Cazenave JP: Stability of the thrombin-thrombomodulin complex on the surface of endothelial cells from human saphenous vein or from the cell line EA.HY 926. Biochem. J. 259: 35–40, 1989PubMedGoogle Scholar
  80. 80.
    Maruyama I, Majerus PW: Protein C inhibitors endocytosis of thrombin-thrombomodulin complexes in A549 lung cancer cells and human umbilical vein endothelial cells. Blood 69: 1481–1484, 1987PubMedGoogle Scholar
  81. 81.
    Moore KL, Andreoli SP, Esmon NL, Esmon CT, Bang NU: Endotoxin enhances tissue factor and suppresses thrombomodulin expression of human vascular endothelium in vitro. J. Clin Invest 79: 124–130, 1987PubMedCrossRefGoogle Scholar
  82. 82.
    Cozzolino F, Torcia M, Miliani A, Carossino AM, Giordani R, Cinotti S, Filimberti E, Saccardi R, Bernabei P, Guidi G et al: Potential role of interleukin-1 as the trigger for diffuse intrasvacular coagulation in acute nonlymphoblastic leukemia. Am. J. Med. 84: 240–250, 1988PubMedCrossRefGoogle Scholar
  83. 83.
    Moore KL, Esmon CT, Esmon NL: Tumor necrosis factor leads to the internalization and degradation of thrombomodulin from the surface of bovine aortic endothelial cells in culture. Blood 73: 159–165, 1989PubMedGoogle Scholar
  84. 84.
    Dittman WA, Kumada T, Sadler JE, Majerus PW: The structure and function of mouse thrombomodulin. Phorbol myristate acetate stimulate degradation and synthesis of thrombomodulin without affecting mRNA levels in hemangioma cells. J. Biol. Chem. 263: 15815–15822, 1988PubMedGoogle Scholar
  85. 85.
    Scarpati EM, Sadler JE: Regulation of endothelial cell coagulant properties. Modulation of tissue factor, Plasminogen activator inhibitors, and thrombomodulin by phorbol 12-myristate 13 acetate and tumor necrosis factor. J. Biol. Chem. 264: 20705–20711, 1989PubMedGoogle Scholar
  86. 86.
    Conway EM, Rosenberg RD: Tumor necrosis factor suppresses transcription of the thrombomodulin gene in endothelial cells. Mol. Cell Biol. 5588–5592, 1989Google Scholar
  87. 87.
    Comp PC, Jacocks RM, Ferrell Gl, Esmon CT: Activation of protein C in vivo. J. Clin Invest. 70: 127–134, 1982PubMedCrossRefGoogle Scholar
  88. 88.
    Ehrlich HJ, Esmon N1, Bang NU: In Vivo behavior of detergent solubilized purified rabbit thrombomodulin on intravenous injection into rabbits. J. Lab. Clin. Med. 115: 182–189, 1990PubMedGoogle Scholar
  89. 89.
    Kumada T, Dittman WA, Majerus PW: A role for thrombomodulin in the pathogenesis of thrombin-induced thromboembolism in mice. Blood 71: 728–733, 1988PubMedGoogle Scholar
  90. 90.
    Gomi K, Aushi M, Honda G, Kawahara S, Matsuzaki O, Kanabayashi T, Yamamoto S, Maruyama I, Suzuki K: Antithrombotic effect of recombinant human thrombomodulin on thrombin-induced thromboembolism in mice. Blood 75: 1396–1399, 1990PubMedGoogle Scholar
  91. 91.
    Freyssinet JM, Wiesel ML, Grunbaum L, Pereillo JM, Gauchy J, Schuhler S, Freund, G, Cazenave JP: Activation of human protein C by blood coagulation factor Xa in the presence of anionic phospholipids. Enhancement by sulphonated polysaccharides. Biochem. J. 261: 341–348, 1989PubMedGoogle Scholar
  92. 92.
    Thompson EA, Salem HH: Factor IXa, Xa, XIa and activated protein C do not have protein C activating ability in the presence of thrombomodulin Thromb. Haemost 59: 339–440, 1988Google Scholar
  93. 93.
    Kisiel W, Canfield WM, Ericsson LH, Davie EW: Anticoagulant properties of bovine plasma Protein C following activation by thrombin. Biochemistry 16: 5824–5830, 1977PubMedCrossRefGoogle Scholar
  94. 94.
    DeWaart PK, Van de Bruis H, Hemker H, Lindhout T: Functional properties of factor Va subunits after proteolytic alterations by activated protein C. Biochim Biophys Acta 799: 38–44, 1984CrossRefGoogle Scholar
  95. 95.
    Guinto ER, Esmon CT: Loss of prothrombin and of factor Xa-factor Va interactions upon inactivation of factor Va by activated protein C. J. Biol. Chem. 259: 13986–13992, 1984PubMedGoogle Scholar
  96. 96.
    Lucklow EA, Lyons DA, Ridgeway TM, Esmon CT, Laue TM: Interaction of clotting factor V heavy chain with prothrombin and prethrombin 1 and role of activated protein C in regulating this interaction Analysis by analytical ultracentrifugation. Biochemistry, 28: 2348–2354, 1989CrossRefGoogle Scholar
  97. 97.
    Nesheim ME, Canfield W, Kisiel W, Mann K: Studies of the capacity of factor Xa to protect factor Va from inactivation by activated protein C. J. Biol. Chem. 257: 1443–1447, 1982PubMedGoogle Scholar
  98. 98.
    Solymoss S, Tucker MM, Tracy PB: Kinetics of inactivation of membrane-bound factor Va by activated protein C. Protein S modulates factor Xa protection, J. Biol. Chem. 263: 14884–14890, 1988PubMedGoogle Scholar
  99. 99.
    Krishnaswamy S, Williams EB, Mann KG: The binding of activated protein C to factors V and Va. J. Biol. Chem. 261: 9684–9693, 1986PubMedGoogle Scholar
  100. 100.
    Marlar R, Kleiss A, Griffin J: Mechanism of action of human activated protein C. A thrombin-dependent anticoagulant protein. Blood 59: 1067–1072, 1982PubMedGoogle Scholar
  101. 101.
    Vehar GA, Davie EW: Preparation and properties of bovine factor VIII (antihemophilic factor). Biochemistry 19: 401–410, 1980PubMedCrossRefGoogle Scholar
  102. 102.
    Walker FJ, Chavin SI, Fay PJ: Inactivation of factor VIII by activated protein C and protein S. Arch. Biochem. Biophys. 252: 322–328, 1987PubMedCrossRefGoogle Scholar
  103. 103.
    Walker FJ, Scandella D, Fay PJ: Identification of the binding site for activated protein C on the light chain of factors V and VIII. J. Biol. Chem. 265: 1484–1489, 1990PubMedGoogle Scholar
  104. 104.
    Koedam JA, Meijers JCM, Sixma JJ, Bouma BN: Inactivation of human factor VIII by activated protein C —cofactor activity of protein S and protective effect of von Willebrand factor. J. Clin Invest, 2: 1236–1243, 1988CrossRefGoogle Scholar
  105. 105.
    Fay PJ, Walker FJ: Inactivation of human factor VIII by activated protein C evidence that factor VIII light chain contains the activated protein C binding site. Biochim. Biophys. Acta. 994: 142–148, 1989PubMedCrossRefGoogle Scholar
  106. 106.
    Fass DN, Hewick RM, Knutson GJ, Nesheim ME, Mann KG: Internal duplication and sequence homology in factors V and VIII. Proc. Natl. Acad. Sci. USA 82: 1688–1691, 1985.PubMedCrossRefGoogle Scholar
  107. 107.
    Walker FJ, Fay PW: Characterization of an interaction between protein C and ceruloplasmin. J. Biol. Chem. 265: 1834–1836, 1990PubMedGoogle Scholar
  108. 108.
    DiScipio RG, Davie EW: Characterization of protein S, a gammacarboxyglutamic acid containing protein from bovine and human plasma. Biochemistry 18: 899–904, 1979PubMedCrossRefGoogle Scholar
  109. 109.
    Stenflo J, Jonsson M: Protein S, a new vitamin K-dependent protein from human plasma. F.E.B.S. Lett. 101: 377–381, 1979CrossRefGoogle Scholar
  110. 110.
    Walker, FJ: The regulation of activated protein C by a new protein A possible function for bovine protein S. J. Biol. Chem. 255: 5521–5524, 1980PubMedGoogle Scholar
  111. 111.
    Schwarz HP, Heeb MJ, Wencel-Drake JD, Griffin JH: Identification and quantitation of protein S in human platelets. Blood 66: 1452–1455, 1985PubMedGoogle Scholar
  112. 112.
    Stern D, Brett J, Harris K, Nawroth P: Participation of endothelial cells in the protein C-protein S anticoagulant pathway the synthesis and release of protein S. J. Cell. Biol. 102: 1971–1978, 1986PubMedCrossRefGoogle Scholar
  113. 113.
    Ploos van Amstel HK, Reitsma PH, Hamulyak K, de Die-Smulders CEM, Mannucci PM, Bertina RM: A mutation in the protein S pseudogene is linked to protein S deficiency in a thrombophilic family. Thromb. Haemost, 62: 897–901, 1989Google Scholar
  114. 114.
    Lundwall A, Dackowski W, Cohen E, Shaffer M, Mahr A, Dahlback B, Stenflo J, Wydro R: Isolation and the sequence of the cDNA for human protein S, a regulator of blood coagulation. Proc. Natl. Acad. Sci. U.S.A. 83: 6716–6720, 1986PubMedCrossRefGoogle Scholar
  115. 115.
    Dahlback B, Lundwall A, Stenflo J: Localization of thrombin cleavage sites in the amino-terminal region of bovine protein S. J. Biol. Chem. 261: 5111–5115, 1986PubMedGoogle Scholar
  116. 116.
    Baker ME, French FS, Joseph DR: Vitamin K-dependent protein S is similar to rat androgen binding protein. Biochem. J. 243: 293–296, 1987PubMedGoogle Scholar
  117. 117.
    Walker FJ: Regulation of activated protein C by protein S The role of phospholipid in factor Va inactivation. J. Biol. Chem. 256: 11128–11131, 1981PubMedGoogle Scholar
  118. 118.
    Walker FJ: Interactions of protein S with membranes. Seminars in thrombosis and hemostasis. 14: 216–222, 1988PubMedCrossRefGoogle Scholar
  119. 119.
    Walker, FJ: Properties of chemically modified protein S effect of the conversion of gamma-carboxy glutamic acid to gamma-methyleneglutamic acid on functional properties. Biochemistry 25: 6305–6311, 1986PubMedCrossRefGoogle Scholar
  120. 120.
    Walker FJ: Regulation of vitamin K-dependent protein S Inactivation by thrombin. J. Biol. Chem. 259: 10335–10339, 1984PubMedGoogle Scholar
  121. 121.
    Dahlback B: Purification of human vitamin K-dependent protein S and its limited proteolysis by thrombin. Biochem. J. 209: 837–846, 1983PubMedGoogle Scholar
  122. 122.
    Dahlback B, Stenflo J: High molecular weight complex in human plasma between vitamin K-dependent protein S and complement component C4b-binding protein. Proc. Natl, Acad. Sci U.S.A., 78: 2512–2516, 1981CrossRefGoogle Scholar
  123. 123.
    Dahlback B: Inhibition of protein Ca cofactor function of human and bovine protein S by C4b-binding protein. J. Biol. Chem. 261: 12022–12027, 1986PubMedGoogle Scholar
  124. 124.
    Comp PC, Doray D, Patton D, Esmon CT: An abnormal plasma distribution of protein S occurs in functional protein S deficiency. Blood 67: 504–508, 1987Google Scholar
  125. 125.
    Hillarp A, Dahlback B: The protein S binding site localized to the central core of C4b-binding protein. J. Biol. Chem. 262: 11300–11307, 1987PubMedGoogle Scholar
  126. 126.
    Walker FJ: Characterization of a synthetic peptide that inhibits the interaction between protein S and C4b-binding protein. J. Biol. Chem. 264: 17645–17649, 1989PubMedGoogle Scholar
  127. 127.
    Comp PC, CT Esmon: Activated protein C inhibits platelet prothrombin converting activity. Blood 54: 1272–1281, 1979PubMedGoogle Scholar
  128. 128.
    Harris KW, Esmon CT: Protein S is required for bovine platelets to support activated protein C binding and activity. J. Biol. Chem. 260: 2007–2010, 1985PubMedGoogle Scholar
  129. 129.
    Suzuki K, Nishioka J, Matsuda M, Murayama H, Hashimoto S, Protein S is essential for the activated protein C-catalyzed inactivation of platelet-associated factor Va. J. Biochem. 96: 455–460, 1984PubMedGoogle Scholar
  130. 130.
    Comp PC, Esmon CT: Generation of fibrinolytic activity by infusion of activated protein C into Dogs. J. Clin. Invest. 68: 1221–1228, 1981PubMedCrossRefGoogle Scholar
  131. 131.
    Seegers W, McCoy LE, Groben HD, Sakuragawa N, Agrawal BBL: Purification and properties of autoprothrombin IIa An anticoagulant perhaps also related to fibrinolysis. Thrombosis Res. 1: 443–448, 1972CrossRefGoogle Scholar
  132. 132.
    Burdick MD, Schaub RG: Human protein C induces anticoagulation and increased fibrinolytic activity in the cat. Thrombosis. Res. 45: 413–419, 1987CrossRefGoogle Scholar
  133. 133.
    Colucci M, Sassen JM, Collen D: Influence of protein C on blood coagulation and fibrinolysis in squirrel monkeys. J. Clin. Invest. 74: 200–202, 1984PubMedCrossRefGoogle Scholar
  134. 134.
    De Fouw NJ, Haverkate F, Bertina RM, Koopman J, Wijngaarden A, van Hinsbergh VWM: The cofactor role of protein S in the acceleration of whole blood clot lysis by activated protein C in vitro. Blood 67: 1189–1195, 1986PubMedGoogle Scholar
  135. 135.
    Comp PC, Esmon CT: in “The Regulation of Coagulation” (eds Mann, K.G and Taylor, F.B.) Elsevier North Holland, Inc pp 583–588, 1980Google Scholar
  136. 136.
    De Fouw NJ, de Jong YF, Haverfkate F, Bertina RM: Activated protein C increases fibrin clot lysis by neutralization of plasminogen activator inhibitor—no evidence for a cofactor role of protein S. Thrombosis and Haemostasis 60: 328–333, 1988PubMedGoogle Scholar
  137. 137.
    Sakata Y, Curriden S, Lawrence D, Griffin JH, Loskutoff DJ: Activated protein C stimulates the fibrinolytic activity of cultured endothelial cells and decreases anti-activator activity. Proc Natl Acad Sci USA 82: 1121–1128 1985PubMedCrossRefGoogle Scholar
  138. 138.
    Fay WP, and Owen WG: Platelet plasminogen activator inhibitor. Purification and characterization of interactions with plasminogen activators and activated protein C. Biochemistry 28: 5773–5778, 1989PubMedCrossRefGoogle Scholar
  139. 139.
    Suzuki K, Nishioka J, Hashimoto S: Protein C inhibitor purification from human plasma and characterization. J. Biol. Chem. 258: 163–168, 1983PubMedGoogle Scholar
  140. 140.
    Kazama Y, Koide T, Saguraga, N: Further characterization of dextran sulfate as a cofactor of protein C inhibition. Thromb. Res. 54: 499–504, 1989PubMedCrossRefGoogle Scholar
  141. 141.
    Heeb MJ, Espana F, Geiger M, Collen D, Stump DK, Griffin JH: Immunological identity of heparin-dependent plasma and urinary protein C inhibitor and plasminogen activator inhibitor-3 J. Biol. Chem. 262: 15813–15816, 1987Google Scholar
  142. 142.
    Espana F, Berrettini M, Griffin JH: Purification and characterization of plasma protein C inhibitor. Thromb. Res. 55: 369–384, 1989PubMedCrossRefGoogle Scholar
  143. 143.
    Pratt, CW, Macik BG, Church, FC: Protein C inhibitor: Purification and proteinase reactivity. Thromb. Res. 53: 595–602, 1989PubMedCrossRefGoogle Scholar
  144. 144.
    Heeb MJ, Bischoll R, Courtney M, Griffin JH: Inhibition of activated protein C by recombinant alpha-1-antitrypsin variants with substitution of arginine for leucine for methionine. J. Biol. Chem. 265: 2365–2369, 1990PubMedGoogle Scholar
  145. 145.
    Heeb MJ, Griffin JH: Physiologic inhibition of human activated protein C by alpha-1-antitrypsin. J. Biol. Chem. 263: 11613–11616, 1988PubMedGoogle Scholar
  146. 146.
    Heeb MJ, Espana F, Griffin JH: Inhibition and complexation of activated protein C by two major inhibitors in plasma. Blood, 73: 446–454, 1989PubMedGoogle Scholar
  147. 147.
    Marlar RA and Kressin DC: Activated protein C is not regulated by alpha-2-macroglobulin. Thrombosis Res. 54: 177–186, 1989CrossRefGoogle Scholar
  148. 148.
    Walker FJ: Protein C in Liver Disease. Annals of Clinical and Laboratory Science 20, 106–112, 1990PubMedGoogle Scholar
  149. 149.
    Vigano S, Mannucci PM, Solinas S. Bottasso B, Mariani G: Decrease in protein C antigen and formation of an abnormal protein soon after starting oral anticoagulant therapy. Br. J. Hematol. 57: 213–220, 1984Google Scholar
  150. 150.
    Rodeghiero F, Mannucci PM, Vigano S, Barbui T, Gugliotta L, Cortellaro M, Dini E: Liver dysfunction rather than intravascular coagulation as the main cause of low protein C and antithrombin III in acute leukemia. Blood, 63: 965–969, 1984PubMedGoogle Scholar
  151. 151.
    Madden RM, Ward M, Marlar RA: Protein C activity levels in endotoxin-induced disseminated intravascular coagulation in a dog model. Thrombosis Res. 55: 297–308, 1989CrossRefGoogle Scholar
  152. 152.
    Taylor FB, Chang A, Esmon CT, D’Angelo S, D’Angelo V, Blick KE: Protein C prevents the coagulopathic and lethal effects of Escherichia coli infusion in the baboon. J. Clin. Invest. 79: 918–925 1987PubMedCrossRefGoogle Scholar
  153. 153.
    Gruber A, Griffin JH, Harker LA, Hanson SR: Inhibition of platelet dependent thrombus formation by human activated protein C in a primate model. Blood 73: 639–642, 1989PubMedGoogle Scholar

Copyright information

© Springer Science+Business Media New York 1991

Authors and Affiliations

  • Frederick J. Walker
    • 1
  1. 1.American Red Cross Blood Services and The Departments of Medicine and Laboratory MedicineUniversity of ConnecticutFarmingtonUSA

Personalised recommendations