The Importance of the Binding of Factor Xa to Phospholipids in the Inhibitory Mechanism of Tissue Factor Pathway Inhibitor: The Transmembrane and Cytoplasmic Domains of Tissue Factor Are not Essential for the Inhibitory Action of Tissue Factor Pathway Inhibitor

Summary

To investigate the inhibitory mechanism of tissue factor pathway inhibitor (TFPI), an attempt was made to examine the inhibitory activity of TFPI toward the factor Vila-truncated tissue factor (TF_1-219) complex, which lacks its transmembrane and cytoplasmic domains. Factor VIIa-TF_1-219 activity was significantly inhibited by TFPI-factor Xa complex in the presence of phospholipids, but was not in the absence of phospholipids. In addition, TFPI did not inhibit factor VIIa-TF_1-219activity in the presence of γ-carboxyglutamic acid-domainless factor Xa. The ability of TFPI-factor Xa complex to inhibit factor VIIa-TF_1-219 activity was totally dependent on the presence of phospholipids and was neutralized by prothrombin fragment 1 in a dose-dependent manner. These results indicate that the transmembrane and cytoplasmic domains of tissue factor are not essential for the inhibitory mechanism of TFPI and confirm that the binding of factor Xa to phospholipids through its γ-carboxyglutamic acid domain is essential for this reaction.

• References

• 1 Davie EW, Fujikawa K, Kisiel W. The coagulation cascade:Initiation maintenance and regulation. Biochemistry 1991; 30: 10363-10370
• 2 Nemerson Y. Tissue factor and hemostasis. Blood 1988; 71: 1-8
• 3 Edgington TS, Mackman N, Brand K, Ruf W. The structural biology of expression and function of tissue factor. Thromb Haemost 1991; 66: 67-79
• 4 Kazama Y, Hamamoto T, Foster DC, Kisiel W. Hepsin a putative membrane-associated serine protease activates human factor VII and initiates a pathway of blood coagulation on the cell surface leading to thrombin formation. J Biol Chem 1995; 270: 66-72
• 5 Rapaport SI. Inhibition of factor VIIa/tissue factor-induced blood coagulation:With particular emphasis upon a factor Xa-dependent inhibitory mechanism. Blood 1989; 73: 359-365
• 6 Broze Jr GJ, Girard TJ, Novotny WF. Regulation of coagulation by a multivalent Kunitz-type inhibitor. Biochemistry 1990; 29: 7539-7546
• 7 Wun T-C, Kretzmer KK, Girard TJ, Miletich JP, Broze Jr G. Cloning and characterization of a cDNA coding for the lipoprotein-associated coagulation inhibitor shows that it consists of three tandem Kunitz-type inhibitory domains. J Biol Chem 1988; 263: 6001-6004
• 8 Girard TJ, Warren LA, Novotny WF, Likert KM, Brown SG, Miletich JP, Broze G. Jr. Functional significance of the Kunitz-type inhibitory domains of lipoprotein-associated coagulation inhibitor Nature 1989; 338: 518-520
• 9 Broze Jr GJ, Warren LA, Novotny WF, Higuchi DA, Girard JJ, Miletich JP. The lipoprotein-associated coagulation inhibitor that inhibits the factor VII-tissue factor complex also inhibits factor Xa:Insight into its possible mechanism of action. Blood 1988; 71: 335-343
• 10 Hamamoto T, Yamamoto M, Nordfang O, Petersen JGL, Foster DC, Kisiel W. Inhibitory properties of full-length and truncated recombinant tissue factor pathway inhibitor (TFPI). Evidence that the third Kunitz-type domain of TFPI is not essential for the inhibition of factor VIIa-tissue factor complexes on cell surfaces. J Biol Chem 1993; 268: 8704-8710
• 11 Kazama Y, Komiyama Y, Kisiel W. Tissue factor pathway inhibitor and protease nexin-1 are major factor Xa binding proteins on the HepG2 cell surface. Blood 1993; 81: 676-682
• 12 Paborsky LR, Caras IW, Fisher KL, Gorman CM. Lipid association but not the transmembrane domain is required for tissue factor activity. Substitution of the transmembrane domain with a phosphatidylinositol anchor. J Biol Chem 1991; 266: 21911-21916
• 13 Ruf W, Kalnik MW, Lund-Hansen T, Edgington TS. Characterization of factor VII association with tissue factor in solution. High and low affinity calcium binding sites in factor VII contribute to functionally distinct interactions. J Biol Chem 1991; 266: 15719-15725
• 14 Waxman E, Ross JBA, Laue TM, Guha A, Thiruvikraman SV, Lin TC, Konigsberg WH, Nemerson Y. Tissue factor and its extracellular soluble domain:The relationship between intermolecular association with factor VIIa and enzymatic activity of the complex. Biochemistry 1992; 31: 3998-4003
• 15 Shigematsu Y, Miyata T, Higashi S, Miki T, Sadler JE, Iwanaga S. Expression of human soluble tissue factor in yeast and enzymatic properties of its complex with factor VIIa. J Biol Chem 1992; 267: 21329-21337
• 16 Ruf W, Rehemtulla A, Edgington TS. Phospholipid-independent and -dependent interactions required for tissue factor receptor and cofactor function. J Biol Chem 1991; 266: 2158-2166
• 17 Yamamoto M, Nakagaki T, Kisiel W. Tissue factor-dependent autoactivation of human blood coagulation factor VII. J Biol Chem 1992; 267: 19089-19094
• 18 Neuenschwander PF, Morrissey JH. Deletion of the membrane anchoring region of tissue factor abolishes autoactivation of factor VII but not cofactor function. Analysis of a mutant with a selective deficiency in activity. J Biol Chem 1992; 267: 14477-14482
• 19 Morrissey JH, Macik BG, Neuenschwander PF, Comp PC. Quantitation of activated factor VII levels in plasma using a tissue factor mutant selectively deficient in promoting factor VII activation. Blood 1993; 81: 734-744
• 20 Kondo S, Kisiel W. Evidence that plasma lipoproteins inhibit the factor VIIa-tissue factor complex by a different mechanism than extrinsic pathway inhibitor. Blood 1987; 70: 1947-1954
• 21 Thim L, Bjoern S, Christensen M, Nicolaisen EM, Lund-Hansen T, Pedersen AH, Hedner U. Amino acid sequence and posttranslational modifications of human factor VIIa from plasma and transfected baby hamster kidney cells. Biochemistry 1988; 27: 7785-7893
• 22 Morita T, Jackson CM. Preparation and properties of derivatives of bovine factor X and factor Xa from which the 7-carboxyglutamic acid containing domain has been removed. J Biol Chem 1986; 261: 4015-4023
• 23 Nordfang O, Bjoern SE, Valentin S, Nielsen LS, Wildgoose P, Beck TC, Hedner U. The C-terminus of tissue factor pathway inhibitor is essential to its anticoagulant activity. Biochemistry 1991; 30: 10371-10376
• 24 Kazama Y, Pastuszyn A, Wildgoose P, Hamamoto T, Kisiel W. Isolation and characterization of proteolytic fragments of human factor VIIa which inhibit the tissue factor-enhanced amidolytic activity of factor VIIa. J Biol Chem 1993; 268: 16231-16240
• 25 Bell WN, Alton HG. A brain extract as a substitute for platelet suspensions in the thromboplastin generation test. Nature 1954; 174: 880-883
• 26 Komiyama Y, Pedersen AH, Kisiel W. Proteolytic activation of human factors IX and X by recombinant human factor VIIa:Effects of calcium phospholipids and tissue factor. Biochemistry 1990; 29: 9418-9425
• 27 Bradford MM. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein dye binding. Anal Biochem 1976; 72: 248-254
• 28 Huang Z-F, Wun T-C, Broze Jr G. Kinetics of factor Xa inhibition by tissue factor pathway inhibitor. J Biol Chem 1993; 268: 26950-26955
• 29 Jesty J, Wun T-C, Lorenz A. Kinetics of the inhibition of factor Xa and the tissue factor-factor VIIa complex by the tissue factor pathway inhibitor in the presence and absence of heparin. Biochemistry 1994; 33: 12686-12694
• 30 Lindhout T, Willems G, Blezer R, Hemker HC. Kinetics of the inhibition of human factor Xa by full-length and truncated recombinant tissue factor pathway inhibitor. Biochem J 1994; 297: 131-136
• 31 Warn-Cramer BJ, Rao LVM, Maki SL, Rapaport SI. Modifications of extrinsic pathway inhibitor (EPI) and factor Xa that affect their ability to interact and to inhibit factor Vila/tissue factor:Evidence for a two-step model of inhibition. Thromb Haemost 1988; 60: 453-456
• 32 Fiore MM, Neuenschwander PF, Morrissey JH. The biochemical basis for the apparent defect of soluble mutant tissue factor in enhancing the proteolytic activities of factor VIIa. J Biol Chem 1994; 269: 143-149
• 33 Valentin S, Nordfang O, Bregengard C, Wildgoose P. Evidence that the C-terminus of tissue factor pathway inhibitor (TFPI) is essential for its in vitro and in vivo interaction with lipoproteins. Blood Coagul Fibrinolysis 1993; 4: 713-720
• 34 Mann KG, Nesheim ME, Church WR, Haley P, Krishnaswamy S. Surface-dependent reactions of the vitamin K-dependent enzyme complexes. Blood 1990; 76: 1-16
• 35 Girard TJ, MacPhail LA, Likert KM, Novotny WF, Miletich JP, Broze Jr G. Inhibition of factor VIIa-tissue factor coagulation activity by a hybrid protein. Nature 1990; 248: 1421-1424