title: Studies of the activation of factor VII bound to tissue factor.
abstract: Experiments were performed to evaluate activation of factor VII bound to relipidated tissue factor (TF) in suspension and to TF constitutively expressed on the surface of an ovarian carcinoma cell line (OC-2008). Activation was assessed by measuring cleavage of 125I-factor VII and by the ability of unlabeled factor VII to catalyze activation of a variant factor IX molecule that, after activation, cannot back-activate factor VII. Factor Xa was found to effectively activate factor VII bound to TF relipidated in either acidic or neutral phospholipid vesicles. Autoactivation of factor VII bound to TF in suspension was dependent on the preparation of TF apoprotein used and the technique of its relipidation. This highlights the need for caution in extrapolating data from TF in suspension to the activation of factor VII bound to cell surfaces during hemostasis. A relatively slow activation of factor VII bound to OC-2008 monolayers in the absence of added protease was observed consistently. Antithrombin in the presence or absence of heparin prevented this basal activation, whereas TF pathway inhibitor (TFPI/factor Xa complexes had only a limited inhibitory effect. Adding a substrate concentration of factor X markedly enhanced basal activation of factor VII, but both TFPI/factor Xa and antithrombin/heparin abolished this enhancement. Overall, our data are compatible with the hypothesis that not all factor VII/TF complexes formed at a site of tissue injury are readily activated to factor VIIa (VIIa)/TF complexes during hemostasis. The clinical significance of this is discussed.
authors.affiliation: Department of Biochemistry, University of Texas Health Center at Tyler 75710, USA.
authors.forename: L V
authors.initials: LV
authors.lastname: Rao
authors.forename: T
authors.initials: T
authors.lastname: Williams
authors.forename: S I
authors.initials: SI
authors.lastname: Rapaport
mesh_terms: D002462:Cell Membrane / Q000378:metabolism; D005167:Factor VII / Q000378:metabolism*; D005260:Female; D006801:Humans; D011485:Protein Binding; D013925:Thromboplastin / Q000378:metabolism*; D014407:Tumor Cells, Cultured
chemical_list: D005167:Factor VII; D013925:Thromboplastin