Download PDF
Thromb Haemost 1993; 70(01): 029-035
DOI: 10.1055/s-0038-1646155
Plenary Lecture
Schattauer GmbH Stuttgart

Molecular Events that Control the Protein C Anticoagulant Pathway

Charles T Esmon
Cardiovascular Biology Research Program, Oklahoma Medical Research Foundation; Departments of Pathology and Biochemistry, University of Oklahoma Health Sciences Center, and Howard Hughes Medical Institute, Oklahoma City, OK, USA
› Author Affiliations
Further Information

Publication History

Publication Date:
24 July 2018 (online)

  PDF Download  Permissions and Reprints
 

  • References

  • 1 Kisiel W. Human plasma protein C: isolation, characterization and mechanism of activation by α-thrombin. J Clin Invest 1979; 64: 761-769
    CrossrefPubMedGoogle Scholar
  • 2 Dittman WA. Thrombomodulin: Biology and potential cardiovascular applications. Trends Cardiovasc Med 1991; 1: 331-336
    CrossrefPubMedGoogle Scholar
  • 3 Esmon CT. The roles of Protein C and thrombomodulin in the regulation of blood coagulation. J Biol Chem 1989; 264: 4743-4746
    PubMedGoogle Scholar
  • 4 Parkinson JF, Koyama T, Bang NU, Preissner KT. Thrombomodulin: An anticoagulant cell surface proteoglycan with physiologically relevant glycosaminoglycan moiety. Adv Exp Med Biol 1992; 313: 177-188
    PubMedGoogle Scholar
  • 5 Gailani D, Broze Jr GJ. Factor XI activation in a revised model of blood coagulation. Science 1991; 253: 909-912
    CrossrefPubMedGoogle Scholar
  • 6 Kurosawa S, Stearns DJ, Jackson KW, 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 1988; 263: 5993-5996
    PubMedGoogle Scholar
  • 7 Stearns DJ, Kurosawa S, Esmon CT. Micro-thrombomodulin: Residues 310-486 from the epidermal growth factor precursor homology domain of thrombomodulin will accelerate protein C activation. J Biol Chem 1989; 264: 3352-3356
    PubMedGoogle Scholar
  • 8 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 1989; 264: 10351-10353
    PubMedGoogle Scholar
  • 9 Parkinson JF, Nagashima M, Kuhn I, Leonard J, Morser J. Structure-function studies of the epidermal growth factor domains of human thrombomodulin . Biochem Biophys Res Commun 1992; 185: 567-576
    CrossrefPubMedGoogle Scholar
  • 10 Grinnell BW, Walls JD, Gerlitz B, Berg DT, McClure DB, Ehrlich H, Bang NU, Yan SB. Native and modified recombinant human protein C: Function, secretion, and posttranslational modifications. In: Protein C and Related Anticoagulants (Advances in Applied Biotechnology Series, vol. 11). Bruley DF, Drohan WN. (eds). Gulf Publishing Company; Houston: 1991. pp. 29-63
    Google Scholar
  • 11 Dolan G, Ball J, Preston FE. Protein C and protein S. Clin Haematol 1989; 2: 999-1042
    PubMedGoogle Scholar
  • 12 Walker FJ, Fay PJ. Regulation of blood coagulation by the protein C system. FASEB J 1992; 6: 2561-2567
    CrossrefPubMedGoogle Scholar
  • 13 Preissner KT. Biological relevance of the protein C system and laboratory diagnosis of protein C and protein S deficiencies. Clin Science 1990; 78: 351-364
    PubMedGoogle Scholar
  • 14 Pabinger I, Brucker S, Kyrle PA, Schneider B, Korninger HC, Niessner H, Lechner K. Hereditary deficiency of antithrombin III, protein C and protein S: prevalence in patients with a history of venous thrombosis and criteria for rational patient screening. Blood Coag Fibrinol 1992; 3: 547-553
    CrossrefPubMedGoogle Scholar
  • 15 Rodgers GM, Chandler WL. Laboratory and clinical aspects of inherited thrombotic disorders. Am J Hematol 1992; 41: 113-122
    CrossrefPubMedGoogle Scholar
  • 16 Comp PC, Esmon CT. Regulatory mechanisms in hemostasis: Natural anticoagulants. In: Hematology: Basic Principles and Practice. Hoffman R, Benz Jr. EJ, Shattil SJ, Furie B, Cohen HJ. (eds). Churchill Livingstone; New York: 1991. pp. 1243-1251
    Google Scholar
  • 17 Stenflo J. Structure-function relationships of epidermal growth factor modules in vitamin K-dependent clotting factors. Blood 1991; 78: 1637-1651
    PubMedGoogle Scholar
  • 18 Davie EW, Fujikawa K, Kisiel W. The coagulation cascade: Initiation, maintenance and regulation. Biochemistry 1991; 30: 10363-10370
    CrossrefPubMedGoogle Scholar
  • 19 Bode W, Turk D, Karshikov A. The refined 1.9-Å X-ray crystal structure of D-Phe-Pro-Arg chloromethylketone-inhibited human alpha-thrombin: Structure analysis, overall structure, electrostatic properties, detailed active-site geometry, and structure-function relationships. Protein Sci 1992; 1: 426-471
    PubMedGoogle Scholar
  • 20 Qiu X, Padmanabhan KP, Carperos VE, Tulinsky A, Kline T, Maraganore JM, Fenton II JW. Structure of the hirulog 3-thrombin complex and nature of the S' subsites of substrates and inhibitors. Biochemistry 1992; 31: 11689-11697
    CrossrefPubMedGoogle Scholar
  • 21 Stubbs MT, Oschkinat H, Mayr I, Huber R, Angliker H, Stone SR, Bode W. The interaction of thrombin with fibrinogen. A structural basis for its specificity. Eur J Biochem 1992; 206: 187-195
    CrossrefPubMedGoogle Scholar
  • 22 Petersen TE. The amino-terminal domain of thrombomodulin and pancreatic stone protein are homologous with lectins. FEBS Lett 1988; 231: 51-53
    CrossrefPubMedGoogle Scholar
  • 23 Ye J, Liu LW, Esmon CT, Johnson AE. The fifth and sixth growth factor-like domains of thrombomodulin bind to the anion exosite of thrombin and alter its specificity. J Biol Chem 1992; 267: 11023-11028
    PubMedGoogle Scholar
  • 24 Suzuki K, Nishioka J, Hayashi T. Localization of thrombomod-ulin-binding site within human thrombin. J Biol Chem 1990; 265: 13263-13267
    PubMedGoogle Scholar
  • 25 Tsiang M, Lentz SR, Dittman WA, Wen D, Scarpati EM, Sadler JE. Equilibrium binding of thrombin to recombinant human thrombomodulin: Effect of hirudin, fibrinogen, factor Va and peptide analogues. Biochemistry 1990; 29: 10602-10612
    CrossrefPubMedGoogle Scholar
  • 26 Hayashi T, Zushi M, Yamamoto S, Suzuki K. Further localization of binding sites for thrombin and protein C in human thrombomodulin. J Biol Chem 1990; 265: 20156-20159
    PubMedGoogle Scholar
  • 27 Bourin MC, Öhlin AK, Lane DA, Stenflo J, Lindahl U. Relationship between anticoagulant activities and polyanionic properties of rabbit thrombomodulin. J Biol Chem 1988; 263: 8044-8052
    PubMedGoogle Scholar
  • 28 Ye J, Esmon CT, Johnson AE. The chondroitin sulfate moiety of thrombomodulin binds a second molecule of thrombin. J Biol Chem 1993; 268: 2373-2379
    PubMedGoogle Scholar
  • 29 Preissner KT, Delvos U, Muller-Berghaus G. Binding of thrombin to thrombomodulin accelerates inhibition of enzyme by antithrombin III.Evidence for a heparin-independent mechanism. Biochemistry 1987; 26: 2521-2528
    CrossrefPubMedGoogle Scholar
  • 30 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 USA 1987; 84: 6425-6429
    CrossrefPubMedGoogle Scholar
  • 31 Wen D, Dittman WA, Ye RD, Deaven LL, Majerus PW, Sadler JE. Human thrombomodulin: Complete cDNA sequence and chromosome localization of the gene. Biochemistry 1987; 26: 4350-4357
    CrossrefPubMedGoogle Scholar
  • 32 Suzuki K, Kusumoto H, Deyashiki Y, Nishioka J, Maruyama I, Zushi M, Kawahara S, Honda G, Yamamoto S, Horiguchi S. Structure and expression of human thrombomodulin, a thrombin receptor on endothelium acting as a cofactor for protein C activation. EMBO J 1987; 6: 1891-1897
    PubMedGoogle Scholar
  • 33 Armstrong SA, Husten EJ, Esmon CT, Johnson AE. The active site of membrane-bound meizothrombin: A fluorescence determination of its distance from the phospholipid surface and its conformational sensitivity to calcium and factor Va. J Biol Chem 1990; 265: 6210-6218
    CrossrefPubMedGoogle Scholar
  • 34 Husten EJ, Esmon CT, Johnson AE. The active site of blood coagulation factor Xa: its distance from the phospholipid surface and its conformational sensitivity to components of the prothrombinase complex. J Biol Chem 1987; 262: 12953-12962
    CrossrefPubMedGoogle Scholar
  • 35 Mutucumarana VP, Duffy EJ, Lollar P, Johnson AE. The active site of factor IXa is located far above the membrane surface and its conformation is altered upon association with factor Villa: A fluorescence study. J Biol Chem 1992; 267: 17012-17021
    PubMedGoogle Scholar
  • 36 Luckow 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 1989; 28: 2348-2354
    CrossrefPubMedGoogle Scholar
  • 37 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 1984; 259: 13986-13992
    PubMedGoogle Scholar
  • 38 Evans DL, Marshall CJ, Christey PB, Carrell RW. Heparin binding site, conformational change, and activation of antithrombin. Biochemistry 1992; 31: 12629-12642
    CrossrefPubMedGoogle Scholar
  • 39 Jakubowski HV, Kline MD, Owen WG. The effect of bovine thrombomodulin on the specificity of bovine thrombin. J Biol Chem 1986; 261: 3876-3882
    PubMedGoogle Scholar
  • 40 Musci G, Berliner LJ, Esmon CT. Evidence for multiple conformational changes in the active center of thrombin induced by complex formation with thrombomodulin: An analysis employing nitroxide spin labels. Biochemistry 1988; 27: 769-773
    CrossrefPubMedGoogle Scholar
  • 41 Ye J, Esmon NL, Esmon CT, Johnson AE. The active site of thrombin is altered upon binding to thrombomodulin: Two distinct structural changes are detected by fluorescence, but only one correlates with protein C activation. J Biol Chem 1991; 266: 23016-23021
    PubMedGoogle Scholar
  • 42 Zushi M, Gomi K, Honda G, Kondo S, Yamamoto S, Hayashi T, Suzuki K. Aspartic acid 349 in the fourth epidermal growth factorlike structure of human thrombomodulin plays a role in its Ca2+-medi-ated binding to protein C. J Biol Chem 1991; 266: 19886-19889
    PubMedGoogle Scholar
  • 43 Le Bonniec BF, MacGillivray RTA, Esmon CT. Thrombin Glu-39 restricts the P'3 specificity to nonacidic residues. J Biol Chem 1991; 266: 13796-13803
    PubMedGoogle Scholar
  • 44 Le Bonniec BF, Esmon CT. Glu-192 [to] Gin substitution in thrombin mimics the catalytic switch induced by thrombomodulin. Proc Natl Acad Sci USA 1991; 88: 7371-7375
    CrossrefPubMedGoogle Scholar
  • 45 Rezaie AR, Esmon CT. The function of calcium in protein C activation by thrombin and the thrombin-thrombomodulin complex can be distinguished by mutational analysis of protein C derivatives. J Biol Chem 1992; 267: 26104-26109
    PubMedGoogle Scholar
  • 46 Ehrlich HJ, Grinnell BW, Jaskunas SR, Esmon CT, Yan SB, Bang NU. Recombinant human protein C derivatives: altered response to calcium resulting in enhanced activation by thrombin. EMBO J 1990; 9: 2367-2373
    PubMedGoogle Scholar
  • 47 Richardson MA, Gerlitz B, Grinnell BW. Enhancing protein C interaction with thrombin results in a clot-activated anticoagulant. Nature 1992; 360: 261-264
    CrossrefPubMedGoogle Scholar
  • 48 Guinto ER, Le Bonniec BF, Esmon CT. The role of motif PPW in thrombin functions. Circulation 1992; 86: 1-412
    CrossrefPubMedGoogle Scholar
  • 49 Le Bonniec BF, Guinto ER, Esmon CT. Interaction of thrombin des-ETW with antithrombin III, the Kunitz inhibitors, thrombomodulin and protein C. J Biol Chem 1992; 267: 19341-19348
    PubMedGoogle Scholar
  • 50 Bode W, Mayr I, Baumann U, Huber R, Stone SR, Hofsteenge J. The refined 1.9 A crystal structure of human a-thrombin: interaction with D-Phe-Pro-Arg chloromethylketone and significance of the Tyr-Pro-Pro-Trp insertion segment. EMBO J 1989; 8: 3467-3475
    PubMedGoogle Scholar
  • 51 Amphlett GW, Kisiel W, Castellino FJ. Interaction of calcium with bovine plasma protein C. Biochemistry 1981; 20: 2156-2161
    CrossrefPubMedGoogle Scholar
  • 52 Esmon NL, DeBault LE, Esmon CT. Proteolytic formation and properties of gamma-carboxyglutamic acid-domainless protein C. J Biol Chem 1983; 258: 5548-5553
    PubMedGoogle Scholar
  • 53 Stearns DJ, Kurosawa S, Sims PJ, Esmon NL, Esmon CT. The interaction of a Ca2+ dependent monoclonal antibody with the protein C activation peptide region: Evidence for obligatory Ca2+ binding to both antigen and antibody. J Biol Chem 1988; 263: 826-832
    PubMedGoogle Scholar
  • 54 Persson E, Selander M, Linse S, Drakenberg T, Öhlin AK, Stenflo J. Calcium binding to the isolated β-hydroxyaspartic acid-containing epidermal growth factor-like domain of bovine factor X. J Biol Chem 1989; 264: 16897-16904
    PubMedGoogle Scholar
  • 55 Ullner M, Selander M, Persson E, Stenflo J, Drakenberg T, Teleman O. Three-dimensional structure of the Apo form of the N-terminal EGF-like module of blood coagulation factor X as determined by NMR spectroscopy and simulated folding. Biochemistry 1992; 31: 5974-5983
    CrossrefPubMedGoogle Scholar
  • 56 Selander-Sunnerhagen M, Ullner M, Persson E, Teleman O, Stenflo J, Drakenberg T. How an epidermal growth factor (EGF)-like domain binds calcium: High resolution NMR structure of the calcium form of the NH2-terminal EGF-like domain in coagulation factor X. J Biol Chem 1992; 267: 19642-19649
    PubMedGoogle Scholar
  • 57 Handford PA, Mayhew M, Baron M, Winship PR, Campbell ID, Brownlee GG. Key residues involved in calcium-binding motifs in EGF-like domains. Nature 1991; 351: 164-167
    CrossrefPubMedGoogle Scholar
  • 58 Öhlin AK, Stenflo J. Calcium-dependent interaction between the EGF region of human protein C and a monoclonal antibody. J Biol Chem 1987; 262: 13798-13804
    PubMedGoogle Scholar
  • 59 Öhlin AK, Linse S, Stenflo J. Calcium binding to the epidermal growth factor homology region of bovine protein C. J Biol Chem 1988; 263: 7411-7417
    PubMedGoogle Scholar
  • 60 Öhlin AK, Landes G, Bourdon P, Oppenheimer C, Wydro R, Stenflo J. β-Hydroxyaspartic acid in the first epidermal growth factor-like domain of protein C: Its role in Ca2+ binding and biological activity. J Biol Chem 1988; 263: 19240-19248
    PubMedGoogle Scholar
  • 61 Sugo T, Bjork I, Holmgren A, Stenflo J. Calcium-binding properties of bovine factor X lacking the gamma-carboxyglutamic acid-containing region. J Biol Chem 1984; 259: 5705-5710
    PubMedGoogle Scholar
  • 62 Rezaie AR, Esmon NL, Esmon CT. The high affinity calcium-binding site involved in protein C activation is outside the first epidermal growth factor homology domain. J Biol Chem 1992; 267: 11701-11704
    PubMedGoogle Scholar
  • 63 Persson E, Bjork I, Stenflo J. Protein structural requirements for Ca2+ binding to the light chain of factor X. Studies using isolated intact fragments containing the gamma-carboxyglutamic acid region and/or the epidermal growth factor-like domains. J Biol Chem 1991; 266: 2444-2452
    PubMedGoogle Scholar
  • 64 Rezaie AR, Neuenschwander PF, Morrissey JH, Esmon CT. Analysis of the functions of the first epidermal growth factor-like domain of factor X. J Biol Chem. 1993 (In Press)
    PubMedGoogle Scholar
  • 65 McCord DM, Monroe DM, Smith KJ, Roberts HR. Characterization of the functional defect in factor IX Alabama: Evidence for a conformational change due to high affinity calcium binding in the first epidermal growth factor domain. J Biol Chem 1990; 265: 10250-10254
    PubMedGoogle Scholar
  • 66 Bourin MC, Lundgren-Åkerlund E, Lindahl U. Isolation and characterization of the glycosaminoglycan component of rabbit thrombomodulin proteoglycan. J Biol Chem 1990; 265: 15424-15431
    PubMedGoogle Scholar
  • 67 Parkinson JF, Vlahos CJ, Yan SCB, Bang NU. Recombinant human thrombomodulin: Regulation of cofactor activity and anticoagulant function by a glycosaminoglycan side chain. Biochem J 1992; 283: 151-157
    CrossrefPubMedGoogle Scholar
  • 68 Bourin MC. Effect of rabbit thrombomodulin on thrombin inhibition by antithrombin in the presence of heparin. Thromb Res 1989; 54: 27-39
    CrossrefPubMedGoogle Scholar
  • 69 Bourin MC, Bjork I, Boffa MC, Lindahl U. Effect of rabbit thrombomodulin on the inhibition of thrombin by antithrombin in the presence of exogenous heparin: Role of the acidic domain of thrombomodulin. Thromb Haemostas 1987; 58: 320 (Abstr.).
    PubMedGoogle Scholar
  • 70 Kurosawa S, Galvin JB, Esmon NL, Esmon CT. Proteolytic formation and properties of functional domains of thrombomodulin. J Biol Chem 1987; 262: 2206-2212
    PubMedGoogle Scholar
  • 71 Parkinson JF, Grinnell BW, Moore RE, Hoskins J, Vlahos CJ, Bang NU. Stable expression of a secretable deletion mutant of recombinant human thrombomodulin in mammalian cells. J Biol Chem 1990; 265: 12602-12610
    PubMedGoogle Scholar
  • 72 Hogg PJ, Öhlin AK, Stenflo J. Identification of structural domains in protein C involved in its interaction with thrombin-thrombomodulin on the surface of endothelial cells. J Biol Chem 1992; 267: 703-706
    PubMedGoogle Scholar
  • 73 Olsen PH, Esmon NL, Esmon CT, Laue TM. The Ca2+-dependence of the interactions between protein C, thrombin and the elastase fragment of thrombomodulin. Analysis by ultracentrifugation. Biochemistry 1992; 31: 746-754
    CrossrefPubMedGoogle Scholar
  • 74 Ruf W, Rehemtulla A, Morrissey JH, Edgington TS. Phospho-lipid-independent and dependent interactions required for tissue factor receptor and cofactor function. J Biol Chem 1991; 266: 2158-2166
    PubMedGoogle Scholar
  • 75 Nesheim ME, Eid S, Mann KG. Assembly of the prothrombinase complex in the absence of prothrombin. J Biol Chem 1981; 256: 9874-9882
    PubMedGoogle Scholar
  • 76 Shigematsu Y, Miyata T, Higashi S, Miki T, Sadler JE, Iwanaga S. Expression of human soluble tissue factor in yeast and enzymatic properties of its complex with factor VIIa. J Biol Chem 1992; 267: 21329-21337
    PubMedGoogle Scholar
  • 77 Higgins DL, Lewis SD, Shafer JA. Steady state kinetic parameters for the thrombin-catalyzed conversion of human fibrinogen to fibrin. J Biol Chem 1983; 258: 9276-9282
    PubMedGoogle Scholar
  • 78 Galvin JB, Kurosawa S, Moore K, Esmon CT, Esmon NL. Reconsti-tution of rabbit thrombomodulin into phospholipid vesicles. J Biol Chem 1987; 262: 2199-2205
    PubMedGoogle Scholar