Plasminogen activator inhibitor-1 inhibits factor VIIa bound to tissue factor

Abstract

Background and objective:A growing body of experimental evidence supports broad inhibitory and regulatory activity of plasminogen activator inhibitor 1 (PAI-1). The present study was designed to investigate whether PAI-1 inhibits factor (F) VIIa complexed with tissue factor (TF), a well-known procoagulant risk factor. Methods and results:The ability of PAI-1 to inhibit FVIIa-TF activity was evaluated in both clotting and factor X (FX) activation assays. PAI-1 and its complex with vitronectin inhibit: (i) clotting activity of FVIIa-TF (PAI-1 IC50, 817 and 125nm, respectively); (ii) FVIIa-TF-mediated FX activation (PAI-1 IC50, 260 and 50nm, respectively); and (iii) FVIIa bound to TF expressed on the surface of stimulated endothelial cells (PAI-1 IC50, 260 and 120nm, respectively). The association rate constant (k a) for PAI-1 inhibition of FVIIa-TF was determined using a chromogenic assay. K a for PAI-1 inhibition of FVIIa bound to relipidated TF is 3.3-fold higher than that for FVIIa bound to soluble TF (k a=0.09±0.01 and 0.027±0.03μm -1min -1, respectively). Vitronectin increases k a for both soluble and relipidated TF by 3.5- and 30-fold, respectively (to 0.094±0.020 and 2.7±0.2μm -1min -1). However, only a 3.5- to 5.0-fold increase in the acylated FVIIa was observed on SDS PAGE in the presence of vitronectin for both relipidated and soluble TF, indicating fast formation of PAI-1/vitronectin/FVIIa/relipidated TF non-covalent complex. Conclusions:Our results demonstrate potential anticoagulant activity of PAI-1 in the presence of vitronectin, which could contribute to regulation of hemostasis under pathological conditions such as severe sepsis, acute lung injury and pleural injury, where PAI-1 and TF are overexpressed. © 2011 International Society on Thrombosis and Haemostasis.