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DOI: 10.1055/s-0038-1648184
Inhibition of Recombinant Human Blood Coagulation Factor VIIa Amidolytic and Proteolytic Activity by Zinc Ions
Publication History
Received 03 September 1990
Accepted after revision 19 December 1990
Publication Date:
24 July 2018 (online)
![](https://www.thieme-connect.de/media/10.1055-s-00035024/199105/lookinside/thumbnails/10-1055-s-0038-1648184-1.jpg)
Summary
Although it is well established that calcium is an essential cofactor in blood coagulation, recent experimental evidence suggests that zinc may also play an important role in hemostasis. In the present study, we have examined the effects of zinc ions on the amidolytic and proteolytic activity of recombinant factor VIIa in the presence of physiological levels of calcium ions. The amidolytic activity of factor VIIa was inhibited half-maximally by 20 ΜM zinc. The amidolytic activity of a derivative of factor VIIa lacking the Γ-carboxyglutamic acid domain was also inhibited half-maximally by 20 ΜM zinc, suggesting that the mechanism of zinc inhibition of factor VIIa amidolytic activity did not involve its Γ-carboxyglutamic acid residues. The amidolytic activity of a complex of recombinant tissue factor and factor VIIa was inhibited half-maximally by 70 ΜM zinc. In contrast to the results obtained with factor VIIa, the amidolytic activities of other human vitamin K-dependent coagulation proteases including factor Xa, thrombin and activated protein C were not appreciably affected by 50-100 ΜM zinc. The proteolytic activation of factor X by a complex of factor VIIa and relipidated tissue factor apoprotein was inhibited half-maximally by 40 ΜM zinc, whereas activation of factor IX in this system was inhibited half-maximally by 70 ΜM zinc ions. Considerably higher levels of zinc (∽100 ΜM) were required to inhibit half-maximally the rate of factor X activation by a complex of factor VIIa and functional tissue factor on the surface of either a human bladder carcinoma cell line, J82, or stimulated human umbilical vein endothelial cells. Activation of factor IX by factor VIIa and tissue factor on the surface of J82 cells was not influenced by zinc. However, the activation rate of factor IX on human umbilical vein endothelial cells was inhibited half-maximally at 100 ΜM zinc. The activation of factor X by factor VIIa in the presence of small unilamellar phospholipid vesicles was inhibited half-maximally by 20 ΜM zinc whereas factor IX activation by factor VIIa was not appreciably influenced by 10-100 ΜM zinc. Our data demonstrate that plasma levels of zinc ions inhibit the amidolytic and proteolytic activities of factor VIIa. The mechanism of this inhibition, as well as its possible physiological relevance, is unknown.
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REFERENCES
- 1 Gordon PR, O’Dell BL. Zinc deficiency and impaired platelet aggregation in guinea pigs. J Nutr 1983; 113: 239-245
- 2 Gordon PR, O’Dell BL. Short-term zinc deficiency and hemostasis in the rat. Proc Soc Exp Biol Med 1980; 163: 240-244
- 3 Marx G, Hopmeier P. Zinc inhibits release of FPA and increases fibrin clot turbidity. Am J Hematol 1986; 22: 347-353
- 4 Greengard JS, Griffin JH. Receptors for high molecular weight kininogen on stimulated washed human platelets. Biochemistry 1984; 23: 6863-6869
- 5 Van Iwaarden F, de Groot PG, Bouma BN. The binding of high molecular weight kininogen to cultured human endothelial cells. J Biol Chem 1988; 263: 4698-4703
- 6 Bjork I, Olson ST, Sheffer RG, Shore JD. Binding of heparin to human high molecular weight kininogen. Biochemistry 1989; 28: 1213-1221
- 7 Greengard JS, Heeb MJ, Ersdal E, Walsh PN, Griffin JH. Binding of coagulation factor XI to washed human platelets. Biochemistry 1986; 25: 3884-3890
- 8 Shimada T, Kato H, Iwanaga S. Accelerating effect of zinc ions on the surface-mediated activation of factor XII and prekallikrein. J Biochem 1987; 102: 913-921
- 9 Aktulga A, Ulatin ON. Normal human platelet zinc content and its release. In: Platelets: Recent Advances in Basic Research and Clinical Aspects Aktulga A. (ed). Excerpta Medica, Amsterdam: 1975: 185-191
- 10 Noguchi M, Sakai T, Kisiel W. Correlation between antigenic and functional expression of tissue factor on the surface of culture human endothelial cells following stimulation by lipopolysaccharide endotoxin. Thromb Res 1989; 55: 87-97
- 11 Kondo S, Kisiel W. Evidence that plasma lipoproteins inhibit the factor VHa-tissue factor complex by a different mechanism than extrinsic pathway inhibitor. Blood 1987; 70: 1947-1954
- 12 Kisiel W. Human plasma protein C: Isolation, characterization and mechanism of activation by Α-thrombin. J Clin Invest 1979; 64: 761-769
- 13 Thim L, Bjoem 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: 7785-7793
- 14 Sakai T, Lund-Hansen T, Thim L, Kisiel W. The carboxyglutamic acid domain of human factor VIIa is essential for its interaction with tissue factor. J Biol Chem 1990; 265: 1890-1894
- 15 Paborsky LR, Tate KM, Harris RJ, Yasura D, Band L, McCray G, Gorman CM, O’Brien DP, Chang JY, Swartz JR, Fung VP, Thomas JN, Vehar GA. Purification of recombinant human tissue factor. Biochemistry 1989; 28: 8072-8077
- 16 Pedersen AH, Nordfang O, Norris F, Wiberg FC, Christensen PM, Moeller KB, Pedersen M, Beck TC, Norris K, Hedner U, Kisiel W. Recombinant human extrinsic pathway inhibitor. Production, isolation and characterization of its inhibitory activity on tissue factor-initiated coagulation reactions. J Biol Chem 1990; 265: 16786-16793
- 17 Bach R, Nemerson Y, Konigsberg W. Purification and characterization of bovine tissue factor. J Biol Chem 1981; 256: 8324-8331
- 18 Bell WN, Alton HG. A brain extract as a substitute for platelet suspensions in the thromboplastin generation test. Nature 1954; 174: 880-001
- 19 Bach R, Gentry R, Nemerson Y. Factor VII binding to tissue factor in reconstituted phospholipid vesicles: Induction of cooperativity by phosphatidylserine. Biochemistry 1986; 25: 4007-4020
- 20 Barenholz Y, Gibbes D, Litman BJ, Goll J, Thompson TE, Carlson FD. A simple method for the preparation of homogeneous phospholipid vesicles. Biochemistry 1977; 16: 2806-2810
- 21 Chen PS, Toribara TY, Warner H. Microdetermination of phosphorus. Analyt Chem 1956; 28: 1756-1758
- 22 Lenz BR, Carpenter TJ, Alford DR. Spontaneous fusion of phosphatidylcholine small unilamellar vesicles in the fluid phase. Biochemistry 1987; 26: 5389-5397
- 23 Griffith MJ, Breikreutz L, Trapp H, Briet E, Noyes CM, Lundblad RL, Roberts HR. Characterization of the clotting activities of structurally different forms of activated factor IX: Enzymatic properties of normal human factor IXaΑ, factor IXa and activated factor IXchapei Hi11 . J Clin Invest 1985; 75: 4-10
- 24 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
- 25 Pasternak CA. A novel form of host defense: Membrane protection by Ca2+ and Zn2+ . Biosci Rep 1987; 7: 81-91
- 26 Sakai T, Lund-Hansen T, Paborsky L, Pedersen AH, Kisiel W. Binding of human factors VII and VIIa to a human bladder carcinoma cell line (J 82): Implications for the initiation of the extrinsic pathway of blood coagulation. J Biol Chem 1989; 264: 9980-9988
- 27 Fair DS, McDonald MJ. Cooperative interaction between factor VII and cell surface expressed tissue factor. J Biol Chem 1987; 262: 11692-11698
- 28 Ploplis VA, Edgington TS, Fair DS. Initiation of the extrinsic pathway of coagulation: Association of factor VIIa with a cell line expressing tissue factor. J Biol Chem 1987; 262: 9503-9508
- 29 Drake TA, Ruf W, Morrissey JH, Edgington TS. Functional tissue factor is entirely cell surface expressed on lipopolysaccharide-stimu-lated human blood monocytes and constitutively tissue factor-producing neoplastic cell line. J Cell Biol 1989; 109: 389-395
- 30 Stem DM, Bank I, Nawroth PP, Cassimeris J, Kisiel W, Fenton JW, Dinarello C, Chess L, Jaffe EA. Self-regulation of procoagulant events on the endothelial cell surface. J Exp Med 1985; 162: 1223-1235
- 31 Moore KL, Andreoli SP, Esmon NL, Esmon CT, Bang NU. Endotoxin enhances tissue factor and suppresses thrombomodulin expression of human vascular endothelium in vitro. J Clin Invest 1987; 79: 124-130
- 32 Silverberg SA, Nemerson Y, Zur M. Kinetics of the activation of bovine coagulation factor X by components of the extrinsic pathway: Kinetic behavior of two-chain factor VII in the presence and absence of tissue factory. J Biol Chem 1977; 252: 8481-8488
- 33 Telgt DSC, Macik BG, McCord DM, Monroe DM, Roberts HR. Mechanism by which recombinant factor VIIa shortens the aPTT: Activation of factor X in the absence of tissue factor. Thromb Res 1989; 56: 603-609
- 34 Rao LVM, Rapaport SI. Factor VIIa-catalyzed activation of factor X independent of tissue factor: Its possible significance for control of hemophilic bleeding by infused factor VIIa. Blood 1990; 75: 1069-1073
- 35 Bom VJJ, Bertina RM. The contributions of calcium, phospholipids and tissue factor apoprotein to the activation of human blood coagulation factor X by activated factor VII. Biochem J 1990; 265: 327-366
- 36 Vallee BL, Auld DS. Zinc coordination, function, and structure of zinc enzymes and other proteins. Biochemistry 1990; 29: 5647-5659
- 37 Liljas A, Kannan KK, Bergsten PC, Waara I, Fridborg B, Strandberg B, Carlbom U, Jarup L, Lovgren S, Petef M. Crystal structure of human carbonic anhydrase C. Nature New Biol 1972; 235: 131-137
- 38 Chapman D, Peel WE, Kingston B, Lilley TH. Lipid phase transitions in model biomembranes: The effect of ions on phosphatidylcholine bilayers. Biochim Biophys Acta 1977; 464: 260-275