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Thromb Haemost 1991; 65(01): 046-051
DOI: 10.1055/s-0038-1647452
Original Article
Schattauer GmbH Stuttgart

Interaction of Plasma Kallikrein with Protein C Inhibitor in Purified Mixtures and in Plasma

Francisco España
The Research Center and Department of Clinical Pathology, Hospital “La Fe”, Valencia, Spain
,
Amparo Estelles
The Research Center and Department of Clinical Pathology, Hospital “La Fe”, Valencia, Spain
,
John H Griffin
*   The Committee on Vascular Biology and Department of Molecular and Experimental Medicine, Research lnstitute of Scripps Clinic, La Jolla, CA, USA
,
Justo Aznar
The Research Center and Department of Clinical Pathology, Hospital “La Fe”, Valencia, Spain
› Author Affiliations
Further Information

Publication History

Received 17 April 1990

Accepted after revision 21 August 1990

Publication Date:
02 July 2018 (online)

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Summary

The interaction between plasma kallikrein (KK) and protein C inhibitor (PCI) and the influence of KK on the complex formation between activated protein C (APC) and PCI was studied in purified systems as well as in plasma in order to assess the significance of these reactions in the plasma milieu. PCI complexed to KK (KK: PCI) or to APC (APC: PCI) was measured by sandwich ELISA’s using antibodies directed against each protein in the complexes. The formation of KK: PCI complexes assayed by this method paralleled the inhibition of KK amidolytic activity by PCI in purified system. Incubation of normal plasma (NHP) at 4 °C, which can induce prekallikrein activation due to cold activation, resulted in PCI inactivation and appearance of KK: PCI complexes. PCI activity fell to 35% of the NHp and 1.2 μ/ml of KK: PCI complex was formed. However, incubation of NHP at room temperature or of prekallikrein deficient plasma at 4 °C did not result in significant decrease of PCI activity. Thus the PCI inactivation was associated with prekallikrein activation and complexation to PCI following cold activation. Incubation of exogenous purified KK with NHP resulted in PCI inactivation and complexation with KK in a temperaturedependent manner. Addition of 2.8 μ/ml KK to plasma at 4 °C resulted in the inactivation of 55% of plasma PCI and the formation of 0.9 μ/ml KK: PCI which represents 21% of the KK added, whereas at 37 °C PCI was inactivated to 30% and only 0.30 μg/ml KK: PCI complexes were measured. These results indicate that PCI is a major KK inhibitor at 4 °C. At 37 °C, PCI accounted for aborfi 7% of the inhibition of the KK added. In separate experiments, following addition of 2.5 μg/ml APC to NHR more than 1 μg/ml of APC: PCI complex was formed in 3 h. When NHP was prior incubated with KK, PCI activity decreased to 10% of that of the normal plasma. Subsequent addition of APC to the plasma treated with KK resulted in formation of only 35 μg/ml of APC: PCI complex compared to 1,350 μg/ml when plasma was not previously incubated with KK. These results indicate that PCI could play a physiological role in the inhibition of plasma KK, and that, in turn, plasma KK can either complex to or inactivate plasma PCI. Thus, KK could modulate the PCI inhibition of APC in plasma.

 

  • References

  • 1 Stenflo J. A new vitamin K-dependent protein: Purification from bovine plasma and preliminary characterization. J Biol Chem 1976; 251: 355-363
    PubMedGoogle Scholar
  • 2 Canfield WK, Nesheim M, Kisiel W, Mann K. Proteolytic inactivation of bovine factor Va by bovine activated protein C. Circulation 1978; 58: 11-210 (Abstr).
    PubMedGoogle Scholar
  • 3 Vehar GA, Davie EW. Preparation and properties of bovine factor VIII. Biochemistry 1980; 19: 401-410
    CrossrefPubMedGoogle Scholar
  • 4 Fulcher CA, Gardiner JE, Griffin JH, Zimmerman TS. Proteolytic inactivation of human factor VIII procoagulant protein by activated human protein C and its analogy with factor V. Blood 1984; 63: 486-489
    CrossrefPubMedGoogle Scholar
  • 5 Branson H, Katz J, Marble R, Griffin JH. Inherited protein C deficiency and a coumarin-responsive chronic relapsing purpura fulminans syndrome in a neonate. Lancet 1983; 2: 1165-1168
    PubMedGoogle Scholar
  • 6 Estelless A, Garcia-Plaza I, Dasi A, Aznar J, Duart M, Sanz G, Perez-Requejo JL, España F, Jimenez C, Abeledo G. Severe inherited “homozygous” protein C deficiency in a newborn infant. Thromb Haemostas 1984; 52: 53-56
    PubMedGoogle Scholar
  • 7 Griffin JH, Evatt B, Zimmerman TS, Kleiss AJ, Wideman C. Deficiency of protein C in congenital thrombotic disease. J Clin Invest 1981; 68: 1370-1373
    CrossrefPubMedGoogle Scholar
  • 8 Bertina RM, Broekmans AW, van der Linden IK, Mertens K. Protein C deficiency in a Dutch family with thrombotic disease. Thromb Haemostas 1982; 48: 1-5
    PubMedGoogle Scholar
  • 9 Suzuki K, Nishioka J, Hashimoto S. Protein C inhibitor. Purification from human plasma and characterization. J Biol Chem 1983; 258: 163-168
    PubMedGoogle Scholar
  • 10 Suzuki K, Nishioka J, Kusumoto H, Hashimoto S. Mechanism of inhibition of activated protein C by protein C inhibitor. J Biochem 1984; 95: 187-195
    CrossrefPubMedGoogle Scholar
  • 11 Suzuki K. Biochemistry and physiology of protein C inhibitor. In: Protein C. Biochemical and Medical Aspects Witt I. (ed). Walter de Gruyter; Berlin: 1985: 43-58
  • 12 España F, Berrettini M, Griffin JH. Purification and characterization of plasma protein C inhibitor. Thromb Res 1989; 55: 369-384
    CrossrefPubMedGoogle Scholar
  • 13 Meijers JC M, Kanters DH A J, Vlooswijk RA A, van Erp HE, Hessing M, Bouma BN. Inactivation of human plasma kallikrein and factor XIa by protein C inhibitor. Biochemistry 1988; 27: 4231-4237
    CrossrefPubMedGoogle Scholar
  • 14 Suzuki K, Deyashiki Y, Nishioka J, Kurachi K, Akira M, Yamamoto S, Hashimoto S. Characterization of a cDNA for human protein C inhibitor. J Biol Chem 1987; 262: 611-616
    PubMedGoogle Scholar
  • 15 Heeb MJ, España F, Geiger M, Collen D, Stump D, Griffin JH. Immunological identity of heparin-dependent plasma and urinary protein C inhibitor and plasminogen activator inhibitor-3. J Biol Chem 1987; 262: 15813-15816
    PubMedGoogle Scholar
  • 16 Stief TW, Radke KP, Heimburger N. Evidence for identity of PCI and plasminogen activator inhibitor 3. Biol Chem Hoppe-Seyler 1987; 368: 1427-1433
    PubMedGoogle Scholar
  • 17 Stump DC, Thienpont M, Collen D. Purification and characterization of a novel inhibitor of urokinase from human urine. Quantitation and preliminary characterization in plasma. J Biol Chem 1986; 261: 12759-12766
    PubMedGoogle Scholar
  • 18 Heeb MJ, Griffin JH. Physiologic inhibition of human activated protein C by alfa 1-antitrypsin. J Biol Chem 1988; 263: 11613-11616
    PubMedGoogle Scholar
  • 19 España F, Griffin JH. Plasma protein C inhibitor (PCI) inhibits procoagulant and profibrinolytic enzymes. Blood 1987; 70: 401 (Abstr).
    PubMedGoogle Scholar
  • 20 Harpel PC, Lewin MF, Kaplan AP. Distribution of plasma kallikrein between Cl Inactivator and α2-macroglobulin in plasma utilizing a new assay for α2-macroglobulin-kallikrein complexes. J Biol Chem 1985; 260: 4257-4263
    PubMedGoogle Scholar
  • 21 Schapira M, Scott CF, Colman RW. Contribution of plasma protease inhibitors to the inactivation of kallikrein in plasma. J Clin Invest 1982; 69: 462-468
    CrossrefPubMedGoogle Scholar
  • 22 van der Graaf F, Koedam JA, Bouma BN. Inactivation of kallikrein in human plasma. J Clin Invest 1983; 71: 149-158
    CrossrefPubMedGoogle Scholar
  • 23 Gruber A, Griffin JH, Harker LA, Hanson SR. Inhibition of platelet-dependent thrombus formation by human activated protein C in a primate model. Blood 1989; 73: 639-642
    PubMedGoogle Scholar
  • 24 España F, Ratnoff OD. The role of prekallikrein and high molecular weight kininogen in the contact activation of Hageman factor (factor-XII) by sulfatides and other agents. J Lab Clin Med 1983; 102: 487-499
    Google Scholar
  • 25 Heeb MJ, Schwarz HP, White T, Lammle B, Berrettini M, Griffin JH. Immunoblotting studies of the molecular forms of protein C in plasma. Thromb Res 1988; 52: 33-43
    CrossrefPubMedGoogle Scholar
  • 26 España F, Griffin JH. Determination of functional and antigen protein C inhibitor and its complexes with activated protein C in plasma by ELISAés. Thromb Res 1989; 55: 671-682
    CrossrefPubMedGoogle Scholar
  • 27 Aznar JA, España F, Aznar J, Tascon A, Jimenez C. Fletcher factor deficiency: Report of a new family. Scand J Haematol 1978; 21: 94-98
    PubMedGoogle Scholar
  • 28 España F, Vicente V, Scharrer I, Tabernero D, Griffin JH. Determination of plasma protein C inhibitor and of two activated protein C inhibitor complexes in normals and in patients with intravas-cular coagulation and thrombotic disease. Thromb Res 1990; 59: 593-608
    CrossrefPubMedGoogle Scholar
  • 29 Aasen AO, Smith ErichsenN, Galhmore MJ, Amundsen E. Studies on components of the plasma kallikrein-kinin system in plasma samples from normal individuals and patients with septic shock. Adv Shock Res 1980; 4: 1-10
    PubMedGoogle Scholar
  • 30 España F, Vicente V, Tabernero D, Scharrer I, Griffin JH. Activated protein C complexes with two physiologic inhibitors: protein C inhibitor and alpha-1-antitrypsin. Thromb Haemostas 1989; 62: 474 (Abstr)
    PubMedGoogle Scholar
  • 31 Gardiner JE, Griffin JH. Studies on human protein C inhibitor in normal and factor V/VIII deficient plasmas. Thromb Res 1984; 36: 197-203
    CrossrefPubMedGoogle Scholar
  • 32 Marlar RA, Griffin JH. Deficiency of protein C inhibitor in combined factor V/VIII deficiency disease. J Clin Invest 1980; 66: 1186-1189
    CrossrefPubMedGoogle Scholar
  • 33 Radke KP, Stief TW, Heimburger N. A new and simple isolation procedure for human protein C inhibitor. Evidence for a second inhibitor for activated protein C present in human plasma. Biol Chem Hoppe-Seyler 1988; 369: 965-974
    PubMedGoogle Scholar
  • 34 Laurell M, Stenflo J. Protein C inhibitor from human plasma: Characterization of native and cleaved inhibitor and demonstration of inhibitor complexes with plasma kallikrein. Thromb Haemostas 1989; 62: 885-891
    PubMedGoogle Scholar
  • 35 Estelles F, Gruber A, Kelly AB, Hanson SR, Harker LA, Griffin JH. In vivo and in vitro complexes of activated protein C with two inhibitors in baboon. Circulation 1988; 78: 318 (Abstr).
    PubMedGoogle Scholar
  • 36 España F, Vicente V, Tabernero D, Griffin JH. Elisa assays for protein C inhibitor (PCI, PAI-3). Heparin-induced decrease in circulating protein C inhibitor. Thromb Haemostas 1989; 62: 383 (Abstr).
    PubMedGoogle Scholar