Heparin Stimulation of the Inhibition of Activated Protein C and other Enzymes by Human Protein C Inhibitor - Influence of the Molecular Weight of Heparin and Ionic Strength
Justo Aznar
1
The Department of Clinical Pathology, “La Fe” University Hospital, Valencia, Spain
,
Francisco España
2
The Research Center, “La Fe” University Hospital, Valencia, Spain
,
Amparo Estellés
2
The Research Center, “La Fe” University Hospital, Valencia, Spain
,
Montserrat Royo
2
The Research Center, “La Fe” University Hospital, Valencia, Spain
The ability of unfractionated (UF) heparin and low-molecular-weight heparin (LMWH) to potentiate the inhibition of fibrinolytic and coagulation factors by protein C inhibitor (PCI) was studied. Inhibition of activated protein C (APC), urokinase plasminogen activator (uPA), tissue plasminogen activator (tPA), thrombin, factor Xa (Xa), factor XIa (XIa) and plasma kallikrein (KK) by PCI was found to be dependent on the size of the polysaccharide. In general, maximal stimulation was reached with UF heparin, except in the case of KK. Differences in heparin stimulation were more pronounced for thrombin, APC, uPA, tPA and XIa, whereas inactivation of Xa by PCI was less dependent on the presence of heparin, and kallikrein showed higher potentiation with LMWH than with UF heparin. The second-order rate constants for enzyme inhibition by PCI were strongly dependent on the ionic strength, and, in general, with an ionic strength higher than 0.15 the heparin stimulation of the inhibition reactions was drastically reduced. These results may explain the large discrepancies in the literature on the effect of heparin on the stimulation of enzyme inhibition by PCI. They also show that LMWH is less efficient in stimulating the PCI inhibition of APC, uPA and tPA, which could contribute to the antithrombotic effect of these enzymes.
References
1
Rosenberg RD.
Actions and interactions of antithrombin and heparin. New Engl J Med 1975; 292: 146-151
2
Rosenberg RD.
The heparin-antithrombin system: a natural anticoagulant mechanism. In: Hemostasis and Thrombosis: Basic Principles and Clinical Practice
Colman RW,
Hirsh J,
Marder VJ,
Salzman EW.
eds
J.B. Lippin-cott Company; Philadelphia, PA: 1987: 1373-1392
3
Andersson LO,
Barrowcliffe TW,
Holmer E,
Johnson EA,
Sims GEC.
Anticoagulant properties of heparin fractionated by affinity chromatography on matrix-bound antithrombin-III and by gel filtration. Thromb Res 1979; 9: 575-583
4
Andersson LO,
Barrowcliffe TW,
Holmer E,
Johnson EA,
Soderstrom G.
Molecular weight dependence of the heparin potentiated inhibition of thrombin and activated factor X. Effect of heparin neutralization in plasma. Thromb Res 1979; 15: 531-541
5
Laurent TC,
Tengblad A,
Thunberg L,
Hook M,
Lindahl U.
On the molecular weight dependence of the anticoagulant activity of heparin. Biochem J 1978; 175: 691-701
6
Johnson EA,
Kirkwood TBL,
Stirling Y,
Perez-Requejo JL.
Four heparin preparations: Anti-Xa potentiating effect of heparin after subcutaneous injection. Thromb Haemost 1976; 35: 586-591
7
Kakkar VV,
Lawrence D,
Bentley PG,
de Haas HA,
Ward VP,
Scully MF.
A comparative study of low doses of heparin and a heparin analogue in the prevention of postoperative deep vein thrombosis. Thromb Res 1978; 13: 111-122
8
Holmer E,
Kurachi K,
Soderstrom G.
The molecular-weight dependence of the rate-enhancing effect of heparin on the inhibition of thrombin, Factor Xa, Factor IXa, Factor Xlla and kallikrein by antithrombin. Biochem J 1981; 193: 395-400
9
Bjork I,
Danielsson A.
Antithrombin and related inhibitors of coagulation proteinases. In: Proteinase Inhibitors
Barrett AS,
Salvesen GS.
eds
Elsevier; Amsterdam: 1986: 489-513
10
Suzuki K,
Nishioka J,
Hashimoto S.
Protein C inhibitor. Purification from human plasma and characterization. J Biol Chem 1983; 258: 163-168
13
Geiger M,
Huber K,
Wojta J,
Stingl L,
España F,
Griffin JH,
Binder BR.
Complex formation between urokinase and plasma protein C inhibitor in vitro and in vivo. Blood 1989; 74: 722-728
14
España F,
Hendl S,
Estellés A,
Fernández PJ,
Gilabert J,
Aznar J,
Griffin JH.
Evidence for the physiologic regulation of urokinase and tissue type plasminogen activators by the serpin, protein C inhibitor, in semen, urine and blood. Thromb Haemost 1993; 70: 989-994
15
España F,
Vicente V,
Vázquez L,
Estellés A,
Sanchez-Cuenca J,
Aznar J.
Activation of protein C and complex formation between protein C inhibitor and tissue plasminogen activator during thrombolytic therapy. Fibrinolysis 1993; 7: 308-315
16
Meijers JCM,
Kanters DHAJ,
Vlooswijk RAA,
van Erp HE,
Hessing M,
Bouma BN.
Inactivation of human plasma kallikrein and factor XIa by protein C inhibitor. Biochemistry 1988; 27: 4231-4237
17
España F,
Estellés A,
Griffin JH,
Aznar J.
Interaction of plasma kallikrein with protein C inhibitor in purified mixtures and in plasma. Thromb Haemost 1991; 65: 46-51
18
Vicente V,
España F,
Tabemero D,
Aznar J,
Estellés A,
Griffin JH.
Evidence of activation of the protein C pathway during acute vascular damage induced by Mediterranean spotted fever. Blood 1991; 78: 416-422
21
España F,
Vicente V,
Tabemero D,
Scharrer I,
Griffin JH.
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. Thromb Res 1990; 59: 593-608
22
Rezaie AR,
Cooper ST,
Church FC,
Esmon CT.
Protein C inhibitor is a potent inhibitor of the thrombin-thrombomodulin complex. J Biol Chem 1995; 270: 25336-25339
24
Hendl S,
España F,
Aznar J,
Estellés A,
Gilabert J,
Griffin JH.
Immuno-affinity purification of protein C with a calcium-dependent monoclonal antibody. Rev Iberoamer Tromb Haemost 1991; 4: 25-28
25
España F,
Estellés A,
Aznar J,
Gilabert J.
Assay of protein C in human plasma: Comparison of amidolytic, coagulation and immunochemical assays. Thromb Res 1986; 44: 771-782
26
van der Graaf F,
Greengard JS,
Bouma BN,
Kerbiriou DM,
Griffin JH.
Isolation and functional characterization of the active light chain of activated human blood coagulation factor XI. J Biol Chem 1983; 258: 9669-9675
27
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
28
España F,
Griffin JH.
Determination of functional and antigen protein C inhibitor and its complexes with activated protein C in plasma by ELISA’s. Thromb Res 1989; 55: 671-682
33
Nurmohamed MT,
Rosendaal FR,
Buller HR,
Dekker E,
Hommes DW,
Vandenbrouke JP,
Briet E.
Low-molecular-weight heparin versus standard heparin in general and orthopaedic surgery: a metaanalysis. Lancet 1992; 340: 152-156
34
Samama MM,
Bara L,
Gerotziafas GT.
Mechanisms for the antithrombotic activity in man of low molecular weight heparins (LMWHs). Haemostasis 1994; 24: 105-117
35
Fernández F,
Nguyen P,
van Ryn J,
Ofosu FA,
Hirsh J,
Buchanan MR.
Hemorrhagic doses of heparin and other glycosaminoglycans induce a platelet defect. Thromb Res 1986; 43: 491-495
36
Blajchman MA,
Young E,
Ofosu FA.
Effects of unfractioned heparin, der-matan sulfate and low molecular weight heparin on vessel wall permeability in rabbits. Ann N Y Acad Sci 1989; 556: 245-254
37
Fareed J,
Walenga M,
Hoppenteadt A,
Messmore L.
Studies on the profibrinolytic actions of heparin and its fractions. In: Progress in Fibrinolysis VII
Davidson JF,
Donati MB,
Coccheri S.
eds
Churchill Livingstone; Edimburg: 1985: 356-367
40
Vinnazer H,
Stemberger A,
Haas S,
Blumel GB.
Influence of heparin, of different heparin fractions and of a low molecular weight heparin-like substance on the mechanism of fibrinolysis. Thromb Res 1982; 27: 341-352
