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

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.

[1]  J. Griffin,et al.  Determination of plasma protein C inhibitor and of two activated protein C-inhibitor complexes in normals and in patients with intravascular coagulation and thrombotic disease. , 1990, Thrombosis research.

[2]  M. Laurell,et al.  Protein C Inhibitor from Human Plasma: Characterization of Native and Cleaved Inhibitor and Demonstration of Inhibitor Complexes with Plasma Kallikrein , 1989, Thrombosis and Haemostasis.

[3]  J. Griffin,et al.  Determination of functional and antigenic protein C inhibitor and its complexes with activated protein C in plasma by ELISA's. , 1989, Thrombosis research.

[4]  J. Griffin,et al.  Purification and characterization of plasma protein C inhibitor. , 1989, Thrombosis research.

[5]  J. Griffin,et al.  Inhibition of platelet-dependent thrombus formation by human activated protein C in a primate model. , 1989, Blood.

[6]  B. Lämmle,et al.  Immunoblotting studies of the molecular forms of protein C in plasma. , 1988, Thrombosis research.

[7]  J. Griffin,et al.  Physiologic inhibition of human activated protein C by alpha 1-antitrypsin. , 1988, The Journal of biological chemistry.

[8]  H. Van Erp,et al.  Inactivation of human plasma kallikrein and factor XIa by protein C inhibitor. , 1988, Biochemistry.

[9]  K. Radtke,et al.  A new and simple isolation procedure for human protein C inhibitor. Evidence for a second inhibitor for activated protein C present in human plasma. , 1988, Biological chemistry Hoppe-Seyler.

[10]  J. Griffin,et al.  Immunological identity of heparin-dependent plasma and urinary protein C inhibitor and plasminogen activator inhibitor-3. , 1987, The Journal of biological chemistry.

[11]  K. Radtke,et al.  Inhibition of urokinase by protein C-inhibitor (PCI). Evidence for identity of PCI and plasminogen activator inhibitor 3. , 1987, Biological chemistry Hoppe-Seyler.

[12]  Koji Suzuki.  Biochemistry and Physiology of Protein C Inhibitor , 1985 .

[13]  J. Griffin,et al.  Studies on human protein C inhibitor in normal and Factor V/VIII deficient plasmas. , 1984, Thrombosis research.

[14]  F. España,et al.  Severe Inherited “Homozygous” Protein C Deficiency in a Newborn Infant , 1984, Thrombosis and Haemostasis.

[15]  J. Griffin,et al.  Proteolytic inactivation of human factor VIII procoagulant protein by activated human protein C and its analogy with factor V. , 1984, Blood.

[16]  J. Nishioka,et al.  Mechanism of inhibition of activated protein C by protein C inhibitor. , 1984, Journal of biochemistry.

[17]  F. España,et al.  The role of prekallikrein and high-molecular-weight kininogen in the contact activation of Hageman factor (factor XII) by sulfatides and other agents. , 1983, The Journal of laboratory and clinical medicine.

[18]  J. Nishioka,et al.  Protein C inhibitor. Purification from human plasma and characterization. , 1983, The Journal of biological chemistry.

[19]  F van der Graaf,et al.  Inactivation of kallikrein in human plasma. , 1983, The Journal of clinical investigation.

[20]  R. Bertina,et al.  Protein C Deficiency in a Dutch Family with Thrombotic Disease , 1982, Thrombosis and Haemostasis.

[21]  R. Colman,et al.  Contribution of plasma protease inhibitors to the inactivation of kallikrein in plasma. , 1982, The Journal of clinical investigation.

[22]  J. Griffin,et al.  Deficiency of protein C in congenital thrombotic disease. , 1981, The Journal of clinical investigation.

[23]  A. Aasen,et al.  Studies on components of the plasma kallikrein-kinin system in plasma samples from normal individuals and patients with septic shock. , 1980, Advances in shock research.

[24]  F. España,et al.  Fletcher factor deficiency: report of a new family. , 2009, Scandinavian journal of haematology.

[25]  J. Stenflo.  A new vitamin K-dependent protein. Purification from bovine plasma and preliminary characterization. , 1976, The Journal of biological chemistry.

[26]  Margaret,et al.  Distribution of Plasma Kallikrein between C i Inactivator and cy 2-Macroglobulin in Plasma Utilizing a New Assay for a 2-Macroglobulin-Kallikrein Complexes * , 2022 .