Download PDF
Thromb Haemost 1998; 80(04): 637-644
DOI: 10.1055/s-0037-1615435
Rapid Communication
Schattauer GmbH

Effect of Phenylglyoxal-modified α2-Antiplasmin on Urokinase-induced Fibrinolysis

Kyung N. Lee
1   From the William K. Warren Medical Research Institute and Department of Medicine, University of Oklahoma Health Sciences Center, Oklahoma City, Oklahoma, USA
,
Steve C. Lee
1   From the William K. Warren Medical Research Institute and Department of Medicine, University of Oklahoma Health Sciences Center, Oklahoma City, Oklahoma, USA
,
Kenneth W. Jackson
1   From the William K. Warren Medical Research Institute and Department of Medicine, University of Oklahoma Health Sciences Center, Oklahoma City, Oklahoma, USA
,
Weon-Chan Tae
1   From the William K. Warren Medical Research Institute and Department of Medicine, University of Oklahoma Health Sciences Center, Oklahoma City, Oklahoma, USA
,
Darlene G. Schwartzott
1   From the William K. Warren Medical Research Institute and Department of Medicine, University of Oklahoma Health Sciences Center, Oklahoma City, Oklahoma, USA
,
Patrick A. McKee
1   From the William K. Warren Medical Research Institute and Department of Medicine, University of Oklahoma Health Sciences Center, Oklahoma City, Oklahoma, USA
› Author Affiliations
Further Information

Publication History

Received 21 August 1997

Accepted after resubmission 19 June 1998

Publication Date:
08 December 2017 (online)

Also available at
    Permissions and Reprints

Summary

One of the functions of activated blood clotting factor XIII (FXIIIa) is the crosslinking of α2-antiplasmin (α2AP) to fibrin. This process results in localization and concentration of α2AP throughout fibrin, thereby making fibrin more resistant to digestion by plasmin. We reasoned that competition by chemically-modified inactive α2AP (modα2AP) with native α2AP would diminish the resistance of fibrin to digestion by plasmin. Modα2AP was prepared by treating native α2AP with an Arg-specific reagent, phenylglyoxal. An average of four of the total nineteen Arg residues in α2AP reacted with phenylglyoxal and resulted in complete loss of plasmin inhibitory activity; however, modα2AP competed effectively with native α2AP for becoming crosslinked to fibrin by FXIIIa catalysis. In the presence of modα2AP, urokinase (UK)-induced plasma clot lysis time shortened significantly. Modα2AP enhanced UK-induced clot lysis in a whole blood system as shown by the similarities of rates of clot lysis for a mixture of 20 U/ml UK and 1.5 μM modα2AP versus that induced by 100 U/ml UK without modα2AP. Less fibrinogenolysis occurred in whole blood when modα2AP was present since much lower UK concentrations were needed to achieve the same level of fibrinolysis than when only native α2AP was present. Our results indicate that modα2AP enhances UK-induced fibrinolysis by competitive inhibition of factor XIIIa-mediated incorporation of native α2AP into fibrin.

 

  • References

  • 1 Tamaki T, Aoki N. Cross-linking of α2-plasmin inhibitor to fibrin catalyzed by activated fibrin-stabilizing factor. J Biol Chem 1982; 257: 14767-72.
    PubMedSearch in Google Scholar
  • 2 Shieh BH, Travis J. The reactive site of human α2-antiplasmin. J Biol Chem 1987; 262: 6055-9.
    PubMedSearch in Google Scholar
  • 3 Wilczynska M, Fa M, Ohlsson PI, Ny T. The inhibition mechanism of ser-pins. Evidence that the mobile reactive center loop is cleaved in the native protease-inhibitor complex. J Biol Chem 1995; 270: 29652-5.
    CrossrefPubMedSearch in Google Scholar
  • 4 Jansen JW, Haverkate F, Koopman J, Nieuwenhuis HK, Kluft C, Boschman TA. Influence of factor XIIIa activity on human whole blood clot lysis in vitro. Thromb Haemost 1987; 57: 171-5.
    Thieme ConnectPubMedSearch in Google Scholar
  • 5 Reed GL III, Matsueda GR, Haber E. Inhibition of clot-bound α2-antiplasmin enhances in vivo thrombolysis. Circulation 1990; 82: 164-8.
    CrossrefPubMedSearch in Google Scholar
  • 6 Sakata Y, Aoki N. Significance of cross-linking of α2-plasmin inhibitor to fibrin in inhibition of fibrinolysis and in hemostasis. J Clin Invest 1982; 69: 536-42.
    CrossrefPubMedSearch in Google Scholar
  • 7 McKee PA, Mattock P, Hill RL. Subunit structure of human fibrinogen, soluble fibrin, and cross-linked insoluble fibrin. Proc Natl Acad Sci USA 1970; 66: 738-44.
    CrossrefPubMedSearch in Google Scholar
  • 8 Shen L, Lorand L. Contribution of fibrin stabilization to clot strength. Supplementation of factorXIII-deficient plasma with the purified zymogen. J Clin Invest 1983; 71: 1336-41.
    CrossrefPubMedSearch in Google Scholar
  • 9 Reed GL 3, Matsueda GR, Haber E. Synergistic fibrinolysis: combined effects of plasminogen activators and an antibody that inhibits α2-antiplasmin. Proc Natl Acad Sci USA 1990; 87: 1114-8.
    CrossrefPubMedSearch in Google Scholar
  • 10 Sakata Y, Eguchi Y, Mimuro J, Matsuda M, Sumi Y. Clot lysis induced by a monoclonal antibody against α2-plasmin inhibitor. Blood 1989; 74: 2692-7.
    PubMedSearch in Google Scholar
  • 11 Weitz JI, Leslie B, Hirsh J, Klement P. α2-antiplasmin supplementation inhibits tissue plasminogen activator-induced fibrinogenolysis and bleeding with little effect on thrombolysis. J Clin Invest 1993; 91: 1343-50.
    CrossrefPubMedSearch in Google Scholar
  • 12 Christensen U, Clemmensen I. Kinetic properties of the primary inhibitor of plasmin from human plasma. Biochem J 1977; 163: 389-91.
    CrossrefPubMedSearch in Google Scholar
  • 13 Wiman B, Collen D. On the mechanism of the reaction between human α2-antiplasmin and plasmin. J Biol Chem 1979; 254: 9291-7.
    PubMedSearch in Google Scholar
  • 14 Marder VJ, Shulman NR. High molecular weight derivatives of human fibrinogen produced by plasmin. II. Mechanism of their anticoagulant activity. J Biol Chem 1969; 244: 2120-4.
    PubMedSearch in Google Scholar
  • 15 Rao AK, Pratt C, Berke A, Jaffe A, Ockene I, Schreiber TL, Bell WR, Knatterud G, Robertson TL, Terrin ML. Thrombolysis in Myocardial Infarction (TIMI) Trial-phase I: hemorrhagic manifestations and changes in plasma fibrinogen and the fibrinolytic system in patients treated with recombinant tissue plasminogen activator and streptokinase. J Am Coll Cardiol 1988; 11: 1-11.
    CrossrefPubMedSearch in Google Scholar
  • 16 Doyle PM, Harris CJ, Carter KR, Simpkin DS, Bailey-Smith P, Stone D, Russell L, Blackwell GJ. Peptides incorporating electrophilic glutamine analogues as potential transglutaminase inhibitors. Biochem Soci Trans 1990; 18: 1318-20.
    PubMedSearch in Google Scholar
  • 17 Freund KF, Doshi KP, Gaul SL, Claremon DA, Remy DC, Baldwin JJ, Pitzenberger SM, Stern AM. Transglutaminase inhibition by 2-[(2-oxopropyl)thio]imidazolium derivatives: mechanism of factor XIIIa inactivation. Biochemistry 1994; 33: 10109-19.
    CrossrefPubMedSearch in Google Scholar
  • 18 Lee KN, Fesus L, Yancey ST, Girard JE, Chung SI. Development of selective inhibitors of transglutaminase. Phenylthiourea derivatives. J Biol Chem 1985; 260: 14689-94.
    PubMedSearch in Google Scholar
  • 19 Leidy EM, Stern AM, Friedman PA, Bush LR. Enhanced thrombolysis by a factor XIIIa inhibitor in a rabbit model of femoral artery thrombosis. Thromb Res 1990; 59: 15-26.
    CrossrefPubMedSearch in Google Scholar
  • 20 Shebuski RJ, Sitko GR, Claremon DA, Baldwin JJ, Remy DC, Stern AM. Inhibition of factor XIIIa in a canine model of coronary thrombosis: effect on reperfusion and acute reocclusion after recombinant tissue-type plasminogen activator. Blood 1990; 75: 1455-9.
    PubMedSearch in Google Scholar
  • 21 Lorand JB, Pilkington TR, Lorand L. Inhibitors of fibrin cross-linking: relevance for thrombolysis. Nature 1966; 210: 1273-4.
    CrossrefPubMedSearch in Google Scholar
  • 22 Gorman JJ, Folk JE. Structural features of glutamine substrates for human plasma factor XIIIa (activated blood coagulation factor XIII). J Biol Chem 1980; 255: 419-27.
    PubMedSearch in Google Scholar
  • 23 Parameswaran KN, Velasco PT, Wilson J, Lorand L. Labeling of ε-lysine crosslinking sites in proteins with peptide substrates of factor XIIIa and transglutaminase. Proc Natl Acad Sci USA 1990; 87: 8472-5.
    CrossrefPubMedSearch in Google Scholar
  • 24 Ichinose A, Tamaki T, Aoki N. Factor XIII-mediated cross-linking of NH2-terminal peptide of α2-plasmin inhibitor to fibrin. FEBS Lett 1983; 153: 369-71.
    CrossrefPubMedSearch in Google Scholar
  • 25 Kimura S, Tamaki T, Aoki N. Acceleration of fibrinolysis by the N-terminal peptide of α2-plasmin inhibitor. Blood 1985; 66: 157-60.
    PubMedSearch in Google Scholar
  • 26 McKee PA, Schwartz ML, Pizzo SV, Hill RL. Cross-linking of fibrin by fibrin-stabilizing factor. Ann N Y Acad Sci 1972; 202: 127-48.
    CrossrefPubMedSearch in Google Scholar
  • 27 Hada M, Kaminski M, Bockenstedt P, McDonagh J. Covalent crosslinking of von Willebrand factor to fibrin. Blood 1986; 68: 95-101.
    PubMedSearch in Google Scholar
  • 28 Bale MD, Westrick LG, Mosher DF. Incorporation of thrombospondin into fibrin clots. J Biol Chem 1985; 260: 7502-8.
    PubMedSearch in Google Scholar
  • 29 Mosher DF. Cross-linking of cold-insoluble globulin by fibrin-stabilizing factor. J Biol Chem 1975; 250: 6614-21.
    PubMedSearch in Google Scholar
  • 30 Mosher DF, Schad PE. Cross-linking of fibronectin to collagen by blood coagulation Factor XIIIa. J Clin Invest 1979; 64: 781-7.
    CrossrefPubMedSearch in Google Scholar
  • 31 Lee SC, Lee KN, Schwartzott DG, Jackson KW, Tae W-C, McKee PA. Purification of human α2-antiplasmin with chicken IgY specific to its carboxy-terminal peptide. Prep Biochem Biotech 1997; 27: 227-37.
    CrossrefPubMedSearch in Google Scholar
  • 32 Folk JE, Chung SI. Transglutaminases. Method Enzymol 1985; 113: 358-75.
    PubMedSearch in Google Scholar
  • 33 Wiman B, Collen D. Purification and characterization of human antiplasmin, the fast-acting plasmin inhibitor in plasma. Eur J Biochem 1977; 78: 19-26.
    CrossrefPubMedSearch in Google Scholar
  • 34 Schwartz ML, Pizzo SV, Hill RL, McKee PA. Human Factor XIII from plasma and platelets. Molecular weights, subunit structures, proteolytic activation, and cross-linking of fibrinogen and fibrin. J Biol Chem 1973; 248: 1395-407.
    PubMedSearch in Google Scholar
  • 35 Lee KN, Birckbichler PJ, Patterson Jr. MK. Colorimetric assay of blood coagulation factor XIII in plasma. Clin Chem 1988; 34: 906-10.
    PubMedSearch in Google Scholar
  • 36 Lee KN, Maxwell MD, Patterson Jr. MK, Birckbichler PJ, Conway E. Identification of transglutaminase substrates in HT29 colon cancer cells: use of 5-(biotinamido)pentylamine as a transglutaminase-specific probe. Biochim Biophys Acta 1992; 1136: 12-6.
    CrossrefPubMedSearch in Google Scholar
  • 37 Hattey E, Wojta J, Binder BR. Monoclonal antibodies against plasminogen and α2-antiplasmin: binding to native and modified antigens. Thromb Res 1987; 45: 485-95.
    CrossrefPubMedSearch in Google Scholar
  • 38 Sumi Y, Ichikawa Y, Nakamura Y, Miura O, Aoki N. Expression and characterization of pro α2-plasmin inhibitor. J Biochem 1989; 106: 703-7.
    CrossrefPubMedSearch in Google Scholar
  • 39 Beebe DP, Aronson DL. An automated fibrinolytic assay performed in microtiter plates. Thromb Res 1987; 47: 123-8.
    CrossrefPubMedSearch in Google Scholar
  • 40 Jones AJ, Meunier AM. A precise and rapid microtitre plate clot lysis assay: methodology, kinetic modeling and measurement of catalytic constants for plasminogen activation during fibrinolysis. Thromb Haemost 1990; 64: 455-63.
    Thieme ConnectPubMedSearch in Google Scholar
  • 41 Urano T, Sakakibara K, Rydzewski A, Urano S, Takada Y, Takada A. Relationships between euglobulin clot lysis time and the plasma levels of tissue plasminogen activator and plasminogen activator inhibitor 1. Thromb Haemost 1990; 63: 82-6.
    Thieme ConnectPubMedSearch in Google Scholar
  • 42 Vermylen C, De Vreker RA, Verstraete M. A rapid enzymatic method for assay of fibrinogen: fibrin polymerization time (FPT test). Clin Chim Acta 1963; 8: 418-24.
    CrossrefPubMedSearch in Google Scholar
  • 43 Tone M, Kikuno R, Kume-Iwaki A, Hashimoto-Gotoh T. Structure of human α2-plasmin inhibitor deduced from the cDNA sequence. J Biochem 1987; 102: 1033-41.
    CrossrefPubMedSearch in Google Scholar
  • 44 Holmes WE, Lijnen HR, Collen D. Characterization of recombinant human α2-antiplasmin and of mutants obtained by site-directed mutagenesis of the reactive site. Biochemistry 1987; 26: 5133-40.
    CrossrefPubMedSearch in Google Scholar
  • 45 Ray Jr. WJ, Koshland Jr. DE. A method for characterizing the type and numbers of groups involved in enzyme action. J Biol Chem 1961; 236: 1973-9.
    PubMedSearch in Google Scholar
  • 46 King MM, Heiny LP. Chemical modification of the calmodulin-stimulated phosphatase, calcineurin, by phenylglyoxal. J Biol Chem 1987; 262: 10658-62.
    PubMedSearch in Google Scholar
  • 47 Abe J, Sidenius U, Svensson B. Arginine is essential for the α-amylase inhibitory activity of the α-amylase/subtilisin inhibitor (BASI) from barley seeds. Biochem J 1993; 293: 151-5.
    CrossrefPubMedSearch in Google Scholar
  • 48 Takahashi K. The reaction of phenylglyoxal with arginine residues in proteins. J Biol Chem 1968; 243: 6171-9.
    PubMedSearch in Google Scholar
  • 49 Kimura S, Aoki N. Cross-linking site in fibrinogen for α2-plasmin inhibitor. J Biol Chem 1986; 261: 15591-5.
    PubMedSearch in Google Scholar
  • 50 Lee KN, Jackson KW, Schwartzott DG, McKee PA. Effects of the carboxy-terminal peptide of α2-antiplasmin on plasminogen activation and fibrinolysis. Circulation 1994; 90: I-619 Abstract 3340.
    PubMedSearch in Google Scholar
  • 51 Lucas MA, Straight DL, Fretto LJ, McKee PA. The effects of fibrinogen and its cleavage products on the kinetics of plasminogen activation by urokinase and subsequent plasmin activity. J Biol Chem 1983; 258: 12171-7.
    PubMedSearch in Google Scholar
  • 52 Stack S, Gonzalez-Gronow M, Pizzo SV. Regulation of plasminogen activation by components of the extracellular matrix. Biochemistry 1990; 29: 4966-70.
    CrossrefPubMedSearch in Google Scholar
  • 53 Verstraete M, Collen D. Thrombolytic therapy in the eighties. Blood 1986; 67: 1529-41.
    PubMedSearch in Google Scholar
  • 54 Sasaki T, Morita T, Iwanaga S. Identification of the plasminogen-binding site of human α2-plasmin inhibitor. J Biochem 1986; 99: 1699-705.
    CrossrefPubMedSearch in Google Scholar
  • 55 Peters RG, Jones WC, Cromartie TH. Inactivation of L-lactate monooxygenase with 2,3-butanedione and phenylglyoxal. Biochemistry 1981; 20: 2564-71.
    CrossrefPubMedSearch in Google Scholar
  • 56 Takata Y, Fujioka M. Chemical modification of arginine residues of rat liver S-adenosylhomocysteinase. J Biol Chem 1983; 258: 7374-8.
    PubMedSearch in Google Scholar
  • 57 Marder VJ. Thrombolytic therapy: overview of results in major vascular occlusions. Thromb Haemost 1995; 74: 101-5.
    Thieme ConnectPubMedSearch in Google Scholar
  • 58 Collen D, Lijnen HR. On the future of thrombolytic therapy for acute myocardial infarction. Am J Cardiol 1993; 72: 46G-50G.
    CrossrefPubMedSearch in Google Scholar
  • 59 Verheugt FW, Meijer A, Lagrand WK, Van Eenige MJ. Reocclusion: the flip side of coronary thrombolysis. J Am Coll Cardiol 1996; 27: 766-73.
    CrossrefPubMedSearch in Google Scholar
  • 60 Lee KN, Tae W-C, Jackson KW, Schwartzott DG, McKee PA. Expression and Characterization of recombinant α2-antiplasmin. FASEB J 1996; 10: A1501 abstract 2890.
    PubMedSearch in Google Scholar
  • 61 Cregg JM, Vedvick TS, Raschke WC. Recent advances in the expression of foreign genes in Pichia pastoris. Bio/Technology 1993; 11: 905-10.
    CrossrefPubMedSearch in Google Scholar