title: Characterization of the interaction of a complex of tissue-type plasminogen activator and plasminogen activator inhibitor type 1 with rat liver cells.
abstract: The present study was undertaken in order to determine the recognition site for tissue-type plasminogen activator-plasminogen activator inhibitor type 1 [t-PA-PAI-1] complexes in rat liver in vivo and in vitro. After intravenous injection into rats t-PA-PAI-1 complexes were rapidly removed from the plasma and the liver took up 80% of the injected dose. Within the liver parenchymal and endothelial liver cells contributed mainly to the uptake of t-PA-PAI-1, and were responsible for 62% and 24% of the liver uptake, respectively. The interaction of t-PA-PAI-1 with isolated rat parenchymal liver cells was of high affinity (Kd 17 nM). A well-known antagonist of the alpha 2-macroglobulin receptor (alpha 2MR/low-density lipoprotein receptor-related protein (LRP), GST-39kDa protein (GST-39kDaP) efficiently inhibited the binding (IC50 0.7 nM) of t-PA-PAI-1 to rat parenchymal liver cells. The interaction of t-PA-PAI-1 with LRP on rat parenchymal liver cells was not Ca2(+)-dependent and is most probably mediated by a specific determinant on PAI-1, since an anti-PAI-1 monoclonal antibody inhibited the binding of t-PA-PAI-1, where as free t-PA did not. The binding of t-PA-PAI-1 to rat hepatocytes could not be inhibited by a complex of plasmin and alpha 2-antiplasmin nor by various other ligands of LRP like beta-VLDL and lactoferrin. Binding of t-PA-PAI-1 to rat parenchymal liver cells was followed by internalization and subsequent degradation in the lysosomal compartment. It is concluded that parenchymal and endothelial liver cells mediate the removal of t-PA-PAI-1 complexes from the circulation. LRP on rat parenchymal liver cells is responsible for the uptake and degradation of t-PA-PAI-1 and may therefore be important for the regulation of the t-PA levels in the circulation.
authors.affiliation: Division of Biopharmaceutics, Leiden Amsterdam Center for Drug Research, University of Leiden, The Netherlands.
authors.forename: J
authors.initials: J
authors.lastname: Kuiper
authors.forename: M
authors.initials: M
authors.lastname: Otter
authors.forename: A H
authors.initials: AH
authors.lastname: Voorschuur
authors.forename: A J
authors.initials: AJ
authors.lastname: van Zonneveld
authors.forename: D C
authors.initials: DC
authors.lastname: Rijken
authors.forename: T J
authors.initials: TJ
authors.lastname: van Berkel
mesh_terms: D000818:Animals; D001692:Biological Transport; D002478:Cells, Cultured; D008099:Liver / Q000502:physiology*; D008297:Male; D017395:Plasminogen Activator Inhibitor 1 / Q000502:physiology*; D011485:Protein Binding; D051381:Rats; D017208:Rats, Wistar; D010959:Tissue Plasminogen Activator / Q000502:physiology*
chemical_list: D017395:Plasminogen Activator Inhibitor 1; D010959:Tissue Plasminogen Activator