title: Catabolism of factor VIIa bound to tissue factor in fibroblasts in the presence and absence of tissue factor pathway inhibitor.
abstract: Vascular injury leads to the exposure of blood to fibroblasts and smooth muscle cells within the vessel wall. These cells constitutively express tissue factor (TF), the cellular receptor for plasma clotting factor VIIa (FVIIa). Formation of TF.FVIIa complexes on cell surfaces triggers the blood coagulation cascade. In the present study, we have investigated the fate of TF.FVIIa complexes formed on the cell surface of fibroblasts in the presence and absence of plasma inhibitor, tissue factor pathway inhibitor (TFPI). FVIIa bound to TF on the cell surface was internalized and degraded without depleting the cell surface TF antigen and activity. TFPI significantly enhanced the TF-specific internalization and degradation of FVIIa. TFPI-enhanced internalization and degradation of FVIIa requires the C-terminal domain of TFPI and factor Xa. TFPI. Xa-mediated internalization of FVIIa was associated with the depletion of TF from the cell surface. A majority of the internalized FVIIa was degraded, but a small portion of the internalized FVIIa recycles back to the cell surface as an intact protein. In addition to TF, other cell surface components, such as low density lipoprotein receptor-related protein (LRP) and heparan sulfates, are essential for TFPI.Xa-induced internalization of FVIIa. Acidification of cytosol, which selectively inhibits the endocytotic pathway via coated pits, inhibited TFPI.Xa-mediated internalization but not the basal internalization of FVIIa. Overall, our data support the concept that FVIIa bound to cell surface TF was endocytosed by two different pathways. FVIIa complexed with TF in the absence of the inhibitor was internalized via a LRP-independent and probably noncoated pit pathway, whereas FVIIa complexed with TF along with the inhibitor was internalized via LRP-dependent coated pit pathway.
authors.affiliation: Department of Biochemistry, The University of Texas Health Center, Tyler, Texas 75708, USA.
authors.forename: A
authors.initials: A
authors.lastname: Iakhiaev
authors.forename: U R
authors.initials: UR
authors.lastname: Pendurthi
authors.forename: J
authors.initials: J
authors.lastname: Voigt
authors.forename: M
authors.initials: M
authors.lastname: Ezban
authors.forename: L
authors.initials: L
authors.lastname: Vijaya Mohan Rao
mesh_terms: D002460:Cell Line; D015942:Factor VIIa / Q000378:metabolism*; D015951:Factor Xa / Q000494:pharmacology; D005347:Fibroblasts / Q000378:metabolism; D006493:Heparin / Q000494:pharmacology; D006801:Humans; D008074:Lipoproteins / Q000494:pharmacology; D026503:Low Density Lipoprotein Receptor-Related Protein-1; D011971:Receptors, Immunologic / Q000502:physiology; D013925:Thromboplastin / Q000502:physiology*
chemical_list: D008074:Lipoproteins; D026503:Low Density Lipoprotein Receptor-Related Protein-1; D011971:Receptors, Immunologic; C051928:lipoprotein-associated coagulation inhibitor; D006493:Heparin; D013925:Thromboplastin; D015942:Factor VIIa; D015951:Factor Xa