Original article| Volume 108, ISSUE 1, P44-52, July 1986

Download started.


Inhibition of one-chain and two-chain forms of human tissue-type plasminogen activator by the fast-acting inhibitor of plasminogen activator in vitro and in vivo

      This paper is only available as a PDF. To read, Please Download here.


      The inhibition of one-chain and two-chain molecular forms of human tissue-type plasminogen activator (t-PA) by the fast-acting inhibitor of plasminogen activator (PA-inhibitor) present in plasma was studied in vitro and in vivo in rabbits. In vitro, both one-chain and two-chain forms of t-PA were neutralized very rapidly in rabbit plasma with high levels of PA-inhibitor. The rate constant of the interaction between two-chain t-PA and PA-inhibitor was estimated to be 3.107 L/mol/ sec. The presence of CNBr-digested fibrinogen, which mimics the effect of fibrin on the activation of plasminogen by t-PA, did not influence the rate constant. Moreover, PA-inhibitor-rich plasma inhibited in a very similar way in vitro thrombolysis by one-chain or two-chain t-PA incorporated into the clot. Injection of one-chain or two-chain t-PA into rabbits with increased levels of PA-inhibitor, induced by endotoxin, resulted in very rapid inhibition of t-PA activity. Within 30 seconds after injection, no residual free t-PA could be demonstrated. Gel filtration analysis showed that the disappearance of t-PA activity was associated with the generation of t-PA-PA-inhibitor complex with an apparent Mr of 100,000. This enzyme-inhibitor complex, like free t-PA, was cleared from the circulation with a half-life of ~2 minutes, mainly via the liver. It is concluded that PA-inhibitor neutralizes one-chain and two-chain molecular forms of t-PA in plasma at very similar rates, both in vitro and in vivo. In this respect, the PA-inhibitor of plasma is different from that isolated from placenta tissue.


      LPS (lipopolysaccharide), Mr (relative molecular mass), PA-inhibitor (fast-acting inhibitor of tissue-type plasminogen activator), t-PA (tissue-type plasminogen activator)
      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 to Translational Research
      Already a print subscriber? Claim online access
      Already an online subscriber? Sign in
      Institutional Access: Sign in to ScienceDirect


        • Collen D
        On the regulation and control of fibrinolysis.
        Thromb Haemost. 1980; 43: 77-89
        • Pennica D
        • Holmes WE
        • Kohr WJ
        • et al.
        Cloning and expression of human tissue-type plasminogen activator cDNA in E. coli.
        Nature. 1983; 301: 214-221
        • Rijken DC
        • Hoylaerts M
        • Collen D
        Fibrinolytic properties of one-chain and two-chain human extrinsic (tissue-type) plasminogen activator.
        J Biol Chem. 1982; 257: 2920-2925
        • Korninger C
        • Stassen JM
        • Collen D
        Turnover of human extrinsic (tissue-type) plasminogen activator in rabbits.
        Thromb Haemost. 1981; 46: 658-661
        • Collen D
        • Stassen JM
        • Verstraete M
        Thrombolysis with human extrinsic (tissue-type) plasminogen activator in rabbits with experimental jugular vein thrombosis.
        J Clin Invest. 1982; 71: 368-376
        • Rånby M
        • Bergsdorf N
        • Nilsson T
        Enzymatic properties of the one- and two-chain form of tissue plasminogen activator.
        Thromb Res. 1982; 27: 175-183
        • Korninger C
        • Collen D
        Neutralization of human extrinsic (tissue-type) plasminogen activator in human plasma: no evidence for a specific inhibitor.
        Thromb Haemost. 1981; 46: 662-665
        • Hoylaerts M
        • Rijken DC
        • Lijnen HR
        • Collen D
        Kinetics of the activation of plasminogen by human tissue plasminogen activator. Role of fibrin.
        J Biol Chem. 1982; 257: 2912-2919
        • Kruithof EKO
        • Tran-Thang C
        • Ransijn A
        • Bachmann F
        Demonstration of a fast acting inhibitor of plasminogen activators in human plasma.
        Blood. 1984; 64: 907-913
        • Chmielewska J
        • Rånby M
        • Wiman B
        Evidence for a rapid inhibitor to tissue plasminogen activator in plasma.
        Thromb Res. 1983; 31: 427-436
        • Juhan-Vague I
        • Moerman B
        • De Cock F
        • Aillaud MF
        • Collen D
        Plasma levels of a specific inhibitor of tissue-type plasminogen activator (and urokinase) in normal and pathological conditions.
        Thromb Res. 1984; 33: 523-530
        • Thorsen S
        • Philips M
        Isolation of tissue-type plasminogen activator-inhibitor complex from human plasma. Evidence for a rapid plasminogen activator inhibitor.
        Biochim Biophys Acta. 1984; 802: 111-118
        • Verheijen JH
        • Chang GTG
        • Kluft C
        Evidence for the occurrence of a fast-acting inhibitor for tissue-type plasminogen activator in human plasma.
        Thromb Haemost. 1984; 51: 392-395
        • Dosne AM
        • Dupuy E
        • Bodevin E
        Production of a fibrinolytic inhibitor by cultured endothelial cells derived from human umbilical vein.
        Thromb Res. 1978; 12: 377-387
        • Emeis JJ
        • Van Hinsbergh WM
        • Verheijen JH
        • Wijngaards G
        Inhibition of tissue-type plasminogen activator by conditioned medium from cultured human and porcine vascular endothelial cells.
        Biochem Biophys Res Commun. 1983; 110: 392-398
        • Loskutoff DJ
        • van Mourik JA
        • Erickson LA
        • Lawrence D
        Detection of an unusually stable fibrinolytic inhibitor produced by bovine endothelial cells.
        in: Proc Natl Acad Sci USA. 80. 1983: 2956-2960
        • Levin EG
        Latent tissue plasminogen activator produced by human endothelial cells in culture: evidence for an enzyme-inhibitor complex.
        in: Proc Natl Acad Sci USA. 80. 1983: 6804-6808
        • Kawano TK
        • Morimoto K
        • Uemura Y
        Urokinase inhibitor in human placenta.
        Nature. 1968; 217: 253-254
        • Kawano T
        • Morimoto K
        • Uemura Y
        Partial purification and properties of urokinase inhibitor from human placenta.
        J Biol Chem. 1970; 67: 333-342
        • Holmberg L
        • Lecander I
        • Persson B
        • Åstedt B
        An inhibitor from placenta specifically binds urokinase and inhibits plasminogen activator released from ovarian carcinoma in tissue culture.
        Biochim Biophys Acta. 1978; 544: 128-137
        • Schleef R
        • Sinha M
        • Erickson LA
        • Loskutoff D
        A quantitative, functional assay for inhibitors of tissue-type plasminogen activator.
        (Abstr 1467) Circulation. 1984; 70: II, 367
        • Lecander I
        • Roblin R
        • Åstedt B
        Differential inhibition of two molecular forms of melanoma cell plasminogen activator by a placental inhibitor.
        Br J Haematol. 1984; 57: 407-412
        • Colucci M
        • Paramo JA
        • Collen D
        Generation in plasma of a fast-acting inhibitor of plasminogen activator in response to endotoxin stimulation.
        J Clin Invest. 1985; 75: 818-824
        • Collen D
        • Rijken DC
        • Van Damme J
        • Billiau A
        Purification of human tissue-type plasminogen activator in centigram quantities from human melanoma cell culture fluid and its conditioning for use in vivo.
        Thromb Haemost. 1982; 48: 294-296
        • Gaffney PJ
        • Curtis AD
        A collaborative study of a proposed international standard for tissue plasminogen activator (t-PA).
        Thromb Haemost. 1985; 53: 134-136
        • Blombäck B
        • Blombäck M
        Purification of human and bovine fibrinogen.
        Ark Kemi. 1956; 10: 415-428
        • Verheijen JH
        • Mullaart E
        • Chang GTG
        • Kluft C
        • Wijngaards G
        A simple sensitive spectrophotometric assay for extrinsic (tissue-type) plasminogen activator applicable to measurements in plasma.
        Thromb Haemost. 1982; 48: 266-269
        • Fraker PJ
        • Speck Jr, JC
        Protein and cell membrane iodinations with a sparingly soluble chloroamide, l,3,4,6-tetrachloro-3a, da-diphenylglycoluril.
        Biochem Biophys Res Commun. 1978; 80: 849-857
        • Lijnen HR
        • Uytterhoeven M
        • Collen D
        Inhibition of trypsin-like proteinases by tripeptide arginyl and lysyl chloromethylketones.
        Thromb Res. 1984; 34: 431-437
        • Astrup T
        • Müllertz S
        Fibrin plate method for estimating fibrinolytic activity.
        Arch Biochem Biophys. 1952; 40: 346-351
        • Rijken DC
        • Collen D
        Purification and characterization of the plasminogen activator secreted by human melanoma cells in culture.
        J Biol Chem. 1981; 256: 7035-7041
        • Rijken DC
        • Juhan-Vague I
        • De Cock F
        • Collen D
        Measurement of human tissue-type plasminogen activator by a two-site immunoradiometric assay.
        J Lab Clin Med. 1983; 101: 274-284
        • van Mourik J
        • Lawrence D
        • Loskutoff DJ
        Purification of an inhibitor of plasminogen activator (antiactivator) synthesized by endothelial cells.
        J Biol Chem. 1984; 259: 14914-14921
        • Fuchs HF
        • Berger H
        • Pizzo SV
        Catabolism of human tissue plasminogen activator in mice.
        Blood. 1985; 65: 539-544