Download PDF
Thromb Haemost 1995; 74(05): 1298-1304
DOI: 10.1055/s-0038-1649930
Original Article
Fibrinolysis
Schattauer GmbH Stuttgart

Characterization of the Interaction of a Complex of Tissue-type Plasminogen Activator and Plasminogen Activator Inhibitor Type 1 with Rat Liver Cells

J Kuiper
1   The Division of Biopharmaceutics, Leiden Amsterdam Center for Drug Research, University of Leiden, Leiden, The Netherlands
,
M Otter
2   The Gaubius Laboratory, TNO Prevention and Health, Leiden, The Netherlands
,
A H Voorschuur
1   The Division of Biopharmaceutics, Leiden Amsterdam Center for Drug Research, University of Leiden, Leiden, The Netherlands
,
A J van Zonneveld
3   The Department of Biochemistry, Amsterdam Medical Center, Amsterdam, The Netherlands
,
D C Rijken
2   The Gaubius Laboratory, TNO Prevention and Health, Leiden, The Netherlands
,
Th J C van Berkel
1   The Division of Biopharmaceutics, Leiden Amsterdam Center for Drug Research, University of Leiden, Leiden, The Netherlands
› Author Affiliations
Further Information

Publication History

Received 30 December 1994

Accepted after revision 02 August 1995

Publication Date:
10 July 2018 (online)

Also available at
  PDF Download  Permissions and Reprints

Summary

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 α2-macroglobulin receptor (α2MR/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 α2-antiplasmin nor by various other ligands of LRP like β-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.

 

  • References

  • 1 Bu G, Warshawsky I, Schwartz AL. Cellular receptors for the plasminogen activators. Blood 1994; 83: 3427-3436
    PubMedGoogle Scholar
  • 2 Otter M, Kuiper J, Berkel ThJ C van, Rijken DC. Mechanisms of tissue- type plasminogen activator clearance by the liver. Ann NY Acad Sci 1992; 667: 431-442
    CrossrefPubMedGoogle Scholar
  • 3 Krause J. Catabolism of tissue-type plasminogen activator, its variants, mutants and hybrids. Fibrinolysis 1988; 2: 133-142
    PubMedGoogle Scholar
  • 4 Kuiper J, Otter M, Rijken DC, van Berkel ThJC. Characterization of the interaction in vivo of tissue-type plasminogen activator with liver cells. J Biol Chem 1988; 263: 18220-18224
    PubMedGoogle Scholar
  • 5 Einarsson M, Smedsrød B, Pertoft H. Uptake and degradation of tissue type plasminogen activator in rat liver. Thromb Haemost 1988; 59: 474-479
    Article in Thieme ConnectPubMedGoogle Scholar
  • 6 Otter M, Zockova P, Kuiper J, van Berkel ThJC, Barrett-Bergshoef MM, Rijken DC. Isolation and characterization of the mannose receptor from human liver potentially involved in the plasma clearance of tissue-type plasminogen activator. Hepatology 1992; 16: 54-59
    CrossrefPubMedGoogle Scholar
  • 7 Otter M, Barrett-Bergshoef MM, Rijken DC. Binding of tissue-type plasminogen activator by the mannose receptor. J Biol Chem 1991; 266: 13931-13935
    PubMedGoogle Scholar
  • 8 Morton PA, Owensby DA, Sobel BE, Schwartz AL. Catabolism of tissue- type plasminogen activator by human hepatoma cell line Hep G2. Modulation by plasminogen activator inhibitor type I J Biol Chem 1989; 264: 7228-7235
    PubMedGoogle Scholar
  • 9 Owensby DA, Morton PA, Schwartz AL. Interactions between tissue-type plasminogen activator and extracellular matrix-associated plasminogen activator inhibitor type 1 in the human hepatoma cell line Hep G2. J Biol Chem 1989; 264: 18180-18187
    PubMedGoogle Scholar
  • 10 Bu G, Maksymovitch EA, Schwartz AL. Receptor mediated endocytosis of tissue-type plasminogen activator by LRP on human hepatoma Hep G2 cells. J Biol Chem 1993; 268: 13002-13008
    PubMedGoogle Scholar
  • 11 Bu G, Williams S, Strickland DK, Schwartz AL. Low density lipoprotein receptor related protein/α2-macroglobulin receptor is a hepatic receptor for tissue-type plasminogen activator. Proc Natl Acad Sci USA 1992; 89: 7427-7431
    CrossrefPubMedGoogle Scholar
  • 12 Orth K, Madison EL, Gething M, Sambrook JF, Herz J. Complexes of tissue-type plasminogen activator and its serpin inhibitor plasminogen activator inhibitor type 1 are internalized by means of the low-density lipoprotein receptor related protein/α2-macroglobulin receptor. Proc Natl Acad Sci USA 1992; 89: 7422-7426
    CrossrefPubMedGoogle Scholar
  • 13 Orth K, Willnow T, Herz J, Gething MJ, Sambrook JF. Low-density lipoprotein receptor-related protein is necessary for the internalization of both tissue-type plasminogen activator-inhibitor complexes and free tissue-type plasminogen activator. J Biol Chem 1994; 269: 21117-21122
    PubMedGoogle Scholar
  • 14 Wing LR, Combe CJ, Bennett B, Booth NA. A receptor on primary human hepatocytes for tissue-type plasminogen activator-plasminogen activator inhibitor type-I and plasmin-α2-antiplasmin complexes. Fibrinolysis 1992; 6: 2
    PubMedGoogle Scholar
  • 15 Combe CJ, Booth NA. Plasmin-α2-macroglobulin complex is not a ligand for the receptor on human hepatocytes for plasmin-α2-antiplasmin complex and tissue-type plasminogen activator in complex with its inhibitor PAI-1. Thromb Haemost 1993; 69: 543
    Article in Thieme ConnectPubMedGoogle Scholar
  • 16 Haijar KA, Reynolds CM. α-fucose-mediated binding and degradation of tissue-type plasminogen activator by HepG2 cells. J Clin Invest 1994; 93: 703-710
    CrossrefPubMedGoogle Scholar
  • 17 Camani C, Bachman F, Kruithof EKO. The role of plasminogen activator inhibitor type-1 in the clearance of tissue-type plasminogen activator by rat hepatoma cells. J Biol Chem 1994; 269: 5770-5775
    PubMedGoogle Scholar
  • 18 Smedsrød B, Einarsson M, Pertoft H. Tissue-type plasminogen activator is endocytosed by mannose and galactose receptors on rat liver cells. Thromb Haemost 1988; 59: 480-484
    Article in Thieme ConnectPubMedGoogle Scholar
  • 19 Otter M, Kuiper J, Bos R, van Berkel ThJC, Rijken DC. Characterization of the interaction both in vivo and in vitro of tissue-type plasminogen activator with rat liver cells. Biochem J 1992; 284: 545-550
    CrossrefPubMedGoogle Scholar
  • 20 Otter M, van Berkel ThJC, Rijken DC. Binding and degradation of tissue- type plasminogen activator by the human hepatoma cell line HepG2. Thromb Haemost 1989; 62: 667-672
    Article in Thieme ConnectPubMedGoogle Scholar
  • 21 Warshawsky I, Bu G, Schwartz AL. 39 kD protein inhibits tissue-type plasminogen activator clearance in vivo. J Clin Invest 1993; 92: 937-944
    CrossrefPubMedGoogle Scholar
  • 22 Smedsrød B, Einarsson M. Clearance of tissue-type plasminogen activator by mannose and galactose receptors in the liver. Thromb Haemost 1990; 63: 60-66
    Article in Thieme ConnectPubMedGoogle Scholar
  • 23 Bakhit C, Lewis D, Billings R, Malfroy B. Cellular catabolism of recombinant tissue-type plasminogen activator. J Biol Chem 1987; 262: 8716-8720
    PubMedGoogle Scholar
  • 24 Ehrlich HJ, Klein Gebbink R, Keijer L, Linders M, Preissner KT, Pannekoek H. Alteration of serpin specificity by a protein cofactor. J Biol Chem 1990; 265: 13029-13035
    PubMedGoogle Scholar
  • 25 Herz J, Goldstein JL, Strickland DK, Ho YK, Brown MS. 39-kDa protein modulates binding of ligands to low-density lipoprotein receptor-related protein/α2-macroglobulin receptor. J Biol Chem 1991; 266: 21232-21238
    PubMedGoogle Scholar
  • 26 Voorschuur AH, Kuiper J, Neelissen JAM, Boers W, van Berkel ThJC. Different zonal distribution of the asialoglycoprotein receptor, the α2-macroglobulin receptor/low-density lipoprotein receptor-related protein and the lipoprotein-remnant receptor of rat liver parenchymal cells. Biochem J 1994; 303: 809-816
    CrossrefPubMedGoogle Scholar
  • 27 Van Berkel ThJC, de Rijke YB, Kruyt JK. Different fate in vivo of oxidatively modified-LDL and acetylated-LDL in rats. Recognition by various scavenger receptors on Kupffer and endothelial liver cells J Biol Chem 1991; 266: 2282-2289
    PubMedGoogle Scholar
  • 28 Kuiper J, Rijken DC, de Munk GA W, van Berkel ThJC. In vivo and in vitro interaction of high- and low-molecular weight single chain urokinase-type plasminogen activator with rat liver cells. J Biol Chem 1992; 267: 1589-1595
    PubMedGoogle Scholar
  • 29 Seglen PO. Isolation of liver cells. Methods Cell Biol 1976; 13: 29-83
    PubMedGoogle Scholar
  • 30 Casteleyn E, van Rooij HC J, van Berkel Th J C, Koster JF. Mechanism of glucagon stimulation of fructose- 1,6-bisphosphate in rat hepatocytes. FEBS Lett 1986; 201: 193-197
    CrossrefPubMedGoogle Scholar
  • 31 Wing LR, Hawksworth GM, Bennet B, Booth NA. Clearance of tissue-type plasminogen activator, PAI-1 and tissue-type plasminogen activator PAI-1 complex in an isolated perfused rat liver system. J Lab Clin Med 1991; 117: 109-114
    PubMedGoogle Scholar
  • 32 Mokuno H, Brady S, Kotice L, Herz J, Havel RJ. Effect of the 39-kDa receptor associated protein on the hepatic uptake and endocytosis of chylomicron remnants and low-density lipoproteins in the rat. J Biol Chem 1994; 269: 13238-13243
    PubMedGoogle Scholar
  • 33 Moestrup SK, Holtet TL, Etzerodt M, Thøgersen HC, Nykjaer A, Andreasen PA, Rasmussen HR, Sottrup-Jensen L, Gliemann J. α2-Macroglobulin-proteinase complexes, plasminogen activator inhibitor type-1-plasminogen activator complexes, and receptor-associated protein bind to a region of the α2-macroglobulin receptor containing a cluster of eight complement-type repeats. J Biol Chem 1993; 268: 13691-13696
    PubMedGoogle Scholar
  • 34 Van Dijk MC M, Kruijt JK, Boers W, Linthorst C, Van Berkel ThJC. Distinct properties of the recognition sites for (3-very low density lipoprotein (remnant receptor) and α2-macroglobulin (low-density lipoprotein receptor- related protein) on rat parenchymal cells. J Biol Chem 1992; 267: 17732-17737
    PubMedGoogle Scholar
  • 35 Williams SE, Ashcom JD, Argraves WS, Strickland DK. A novel mechanism for controlling the activity of α2-macroglobulin receptor/low density lipoprotein receptor-related protein. J Biol Chem 1992; 267: 9035-9040
    CrossrefPubMedGoogle Scholar
  • 36 Moestrup SK, Gliemann J. Analysis of ligand recognition by the purified a2-macroglobulin receptor (low-density lipoprotein receptor-related protein). J Biol Chem 1991; 266: 14011-14017
    PubMedGoogle Scholar
  • 37 Chan AL, Morris HR, Panico M, Etienne AT, Rogers ME, Gaffney P, Creighton-Kempsford L, Dell A. A novel sialylated N-acetylgalactos-amine-containing oligosaccharide is the major complex-type structure present in Bowes melanoma tissue plasminogen activator. Glycobiol 1991; 1: 173-185
    CrossrefPubMedGoogle Scholar