Binding of EGF1 domain peptide in coagulation factor VII with tissue factor and its implications for the triggering of coagulation

Abstract

The binding function of EGF1 domain peptide with tissue factor (TF) and its ability of triggering coagulation were explored. The TF expression model in vitro was established by lipopolysaccharide induction. The affinity of EGFP-EGF1 and TF expressing cells was analyzed by fluorescence microscopy and flow cytometry (FCM). The affinity of EGFP-EGF1 and rat soluble TF was quantitated by surface plasmon resonance (SPR). The ability of EGFP-EGF1 in triggering coagulation was tested by prothrombin time assay. The FCM results showed recombinant factor VII (rFVII) could definitely depress the integration of EGFP-EGF1 with recombinant TF (rTF) (68.65%±3.86% vs 57.98%±4.71%, P<0.01). The SPR results indicated the association constant ka of EGFP-EGF1 proteins was higher than rFVII (8.29±1.39 vs 3.75±0.32, P<0.01). However, the EGFP-EGF1 protein lost the activity of triggering coagulation as compared with blood plasma of normal SD rats (56.8±3.2 s vs 17.8±3.4 s, P<0.01). It was concluded that the rat EGF1 peptide could specifically bind to TF without the ability of triggering coagulation. EGF1 peptide may be a good target head for delivering drugs to TF in anticoagulation therapy.

Keywords

coagulation factor VII / epidermal growth factor-like domain / tissue factor / anticoagulation

References

[1] MackmanN., TilleyR.E., KeyN.S.. Role of the extrinsic pathway of blood coagulation in hemostasis and thrombosis. Arterioscler Thromb Vasc Biol, 2007, 27(8): 1687-1693

[2] JulianN.. Tissue-factor-endowed leukocytes do cause thrombosis. Blood, 2005, 106(4): 1506-1507

[3] SharleneM.D., JenniferL.R., BrianP., et al.. Macrovascular thrombosis is driven by tissue factor derived primarily from the blood vessel wall. Blood, 2005, 105(1): 192-198

[4] OrfeoT., ButenasS., Brummel-ZiedinsK.E., et al.. The tissue factor requirement in blood coagulation. J Biol Chem, 2005, 280(52): 42 887-42 896

[5] ZhangX., HuY., HongM., et al.. Plasma thrombomodulin, fibrinogen, and activity of tissue factor as risk factors for acute cerebral infarction. Am J Clin Pathol, 2007, 128(2): 287-292

[6] LeviM., van der PollT., BüllerH.R.. Bidirectional relation between inflammation and coagulation. Circulation, 2004, 109(22): 2698-2704

[7] MackmanN.. Role of tissue factor in hemostasis, thrombosis, and vascular development. Arterioscler Thromb, 2004, 24(6): 1015

[8] VanA.C., LiX., BoerO.J.. Tissue factor expression in the morphologic spectrum of vulnerable atherosclerotic plaques. Semin Thromb Hemost, 2006, 32(1): 40-47

[9] HeW.J., HuY., ZhangX.P., et al.. Activated protein C ratio, plasma tissue factor activity and activated factor VII in Chinese patients with coronary heart disease. Eur J Med Res, 2008, 13(2): 47-51

[10] DelGiudiceL.A., WhiteG.A.. The role of tissue factor and tissue factor pathway inhibitor in health and disease states. J Vet Emerg Crit Care (San Antonio), 2009, 19(1): 23-29

[11] LeviM.. Disseminated intravascular coagulation: What’s new?. Crit Care Clin, 2005, 21(3): 449-467

[12] LangerF., SpathB., HauboldK., et al.. Tissue factor procoagulant activity of plasma microparticles in patients with cancer-associated disseminated intravascular coagulation. Ann Hematol, 2008, 87(6): 451-457

[13] LwaleedB.A., CooperA.J., VoegeliD., et al.. Tissue factor: a critical role in inflammation and cancer. Biol Res Nurs, 2007, 9(2): 97-107

[14] HagenF.S., GrayC.L., O’HaraP., et al.. Characterization of a cDNA encoding for human FVII. Proc Natl Acad Sci, 1986, 83(8): 2412-2416

[15] AshileyC.P., AndrzejM.B., ShirleyM.R., et al.. Structure of human factor VIIa and its implications for the triggering of blood coagulation. Proc Natl Acad Sci, 1999, 96(16): 8925-8930

[16] ShobhaS., KathleenM., CharityA., et al.. Cloning and expression of rat coagulation factor VII. Thromb Res, 2003, 109(4): 225-231

[17] MayH., HuY., ZhangY.H., et al.. Expression of fusion protein encoding EGFP-EGF1 of rat coagulation factor VII and its binding function. Zhongguo Shi Yan Xue Ye Xue Za Zhi (Chinese), 2008, 16(1): 181-184

[18] BerndE., SusanneZ., KorbinianB., et al.. Tissue factor expression of human monocytes is suppressed by lysophosphatidylcholine. Arterioscler Thromb Vasc Biol, 1999, 19(1): 47-53

[19] KougiasP., ChaiH., LinP.H., et al.. Lysophosphatidylcholine and secretory phospholipase A2 in vascular disease: mediators of endothelial dysfunction and atherosclerosis. Med Sci Monit, 2006, 12(1): RA5-16

[20] VilesJ.F., AnandS.X., ZafarM.U., et al.. Tissue factor coagulation pathway: a new therapeutic target in atheroth-rombosis. J Cardiovasc Pharmacol, 2004, 43(5): 669-676

[21] JamesH.M.. Tissue factor: a key molecule in hemostatic and nonhemostatic systems. Inter J Hemat, 2004, 79(2): 103-108

[22] EilertsenK.E., OsterudB.. Tissue factor: (patho)physiology and cellular biology. Blood, 2004, 15(7): 521-538

[23] HutterR., ValdiviezoC., SauterB.V., et al.. Caspase-3 and tissue factor expression in lipid-rich plaque macrophages. Circulation, 2004, 109(16): 2001-2008

[24] SteffelJ., LüscherT.F., TannerF.C.. Tissue factor in cardiovascular diseases molecular mechanisms and clinical implications. Circulation, 2006, 113(5): 722-731

[25] LeeA., AgnelliG., BullerH., et al.. Dose-response study of recombinant factor VIIa/tissue factor inhibitor recombinant nematode anticoagulant protein c2 in prevention of postoperative venous thromboembolism in patients undergoing total knee replacement. Circulation, 2001, 104(1): 74-78

[26] HimberJ., WohlgensingerC., RouxS., et al.. Inhibition of tissue factor limits the growth of venous thrombus in the rabbit. J Thromb Haemost, 2003, 1(5): 889-895

[27] KasthuriR.S., TaubmanM.B., MackmanN.. Role of tissue factor in cancer. J Clin Oncol, 2009, 27(29): 4834-4838

[28] MilsomC.C., YuJ.L., MackmanN., et al.. Tissue factor regulation by epidermal growth factor receptor and epithelial-to-mesenchymal transitions: effect on tumor initiation and angiogenesis. Cancer Res, 2008, 68(24): 10 068-10 076

[29] IlliamsonV., PykeA., SridharaS., et al.Interspecies exchange mutagenesis of the first epidermal growth factor-like domain of human factor VII, 2005, 3(6): 1250-1256

[30] RobertA.L., AlanG.O., CharlesE., et al.. Inhibitors of tissue factor-factor VIIa for anticoagulant therapy. Curr Med Chem, 2004, 11(17): 2275-2290

[31] ThiecF., CherelG., ChristopheO.D., et al.. Role of the Gla and first epidermal growth factor-like domains of factor X in the prothrombinase and tissue factor-factor VIIa complexes. J Biol Chem, 2003, 278(12): 10 393-10 399

[32] RichR.L., HothL.R., GeogheganK.F., et al.. Kinetic analysis of estrogen receptor/ligand interactions. Proc Natl Acad Sci, 2002, 99(13): 8562-8567

[33] HockalingamK., ChenZ., KatzenellenbogenJ.A., et al.. Directed evolution of specific receptor-ligand pairs for use in the creation of gene switches. Proc Natl Acad Sci, 2005, 102(16): 5691-5696

[34] HuberW., MuellerF.. Biomolecular interaction analysis in drug discovery using surface plasmon resonance technology. Curr Pharm Des, 2006, 12(11): 3999-4021

[35] TaniousF.A., NguyenB., WilsonW.D.. Biosensor-surface plasmon resonance methods for quantitative analysis of biomolecular interactions. Methods Cell Biol, 2008, 84: 53-77