PDF herunterladen
Thromb Haemost 2003; 89(01): 65-73
DOI: 10.1055/s-0037-1613544
Blood Coagulation, Fibrinolysis and Cellular Haemostasis
Schattauer GmbH

Localization of tissue factor pathway inhibitor to lipid rafts is not required for inhibition of factor VIIa/tissue factor activity

Dennis J. Dietzen
1   Department of Pediatrics, Washington University, St. Louis, Missouri
,
Garnet G. Jack
3   Pathology and Research Services, VA Medical Center, Memphis, Tennessee
,
Keith L. Page
1   Department of Pediatrics, Washington University, St. Louis, Missouri
,
Tina A. Tetzloff
1   Department of Pediatrics, Washington University, St. Louis, Missouri
,
Connie L. Hall
2   Pritzker Institute of Medical Engineering, Illinois Institute of Technology, Chicago, Illinois
,
Alan E. Mast
3   Pathology and Research Services, VA Medical Center, Memphis, Tennessee
4   Department of Pathology, University of Tennessee, Memphis, Tennessee, USA
› Institutsangaben
Weitere Informationen

Publikationsverlauf

Received 02. Juli 2002

Accepted after resubmission 25. Oktober 2002

Publikationsdatum:
09. Dezember 2017 (online)

    Lizenzen und Reprints

Summary

Tissue factor pathway inhibitor (TFPI) abrogates coagulation initiated by the factor VIIa/tissue factor catalytic complex. While the gene structure of TFPI suggests that it is a secreted protein, a large pool of TFPI is associated with the vascular endothelium through its affinity for a glycosylphosphatidylinositol (GPI)-linked membrane protein. Inhibition of tissue factor by TFPI coincides with the translocation of quaternary complexes containing tissue factor, factor VIIa, factor Xa, and TFPI to detergent-insoluble plasma membrane domains rich in cholesterol, sphingomyelin, and GPI-linked proteins known as lipid rafts and caveolae. It is not known if localization of TFPI to these membrane domains is required for its inhibition of tissue factor procoagulant activity. We generated chimeric TFPI molecules linked directly to the plasma membrane via a GPI anchor or hydrophobic transmembrane domain and expressed these in HEK293 cells that produce tissue factor but not endogenous TFPI. The GPI-anchored chimera was exclusively enriched in detergent-insoluble membrane fractions while the transmembrane molecule was not. Transfectants expressing equal levels of the GPI-linked or transmembrane TFPI displayed equal anticoagulant potency as assessed by tissue factor-mediated conversion of factor X to factor Xa. Disruption of lipid rafts with cyclodextrin likewise had no effect on the inhibitory activity of the transmembrane or GPI-linked TFPI chimeras in HEK293 cells, nor on endogenous TFPI expressed by ECV304 cells. Thus, we conclude that the GPI anchor and membrane localization to lipid rafts does not enhance inhibition of factor VIIa/ tissue factor by cell-surface associated TFPI.

Keywords

Tissue factor pathway inhibitor - glycosyl phosphatidylinositol - lipid rafts - coagulation - endothelikum
 

  • References

  • 1 Mackman N, Morrissey JH, Fowler B, Edgington TS. Complete sequence of the human tissue factor gene, a highly regulated cellular receptor that initiates the coagulation protease cascade. Biochemistry 1989; 28 (04) 1755-62.
    CrossrefPubMedGoogle Scholar
  • 2 Novotny WF, Girard TJ, Miletich JP, Broze Jr GJ. Purification and characterization of the lipoprotein-associated coagulation inhibitor from human plasma. J Biol Chem 1989; 264 (31) 18832-7.
    PubMedGoogle Scholar
  • 3 Girard TJ, Warren LA, Novotny WF, Likert KM, Brown SG, Miletich JP. et al. Functional significance of the Kunitz-type inhibitory domains of lipoprotein-associated coagulation inhibitor. Nature 1989; 338 6215 518-20.
    CrossrefPubMedGoogle Scholar
  • 4 Lockett JM, Mast AE. Contribution of regions distal to glycine-160 to the anticoagulant activity of tissue factor pathway inhibitor. Biochemistry 2002; 41 (15) 4989-97.
    CrossrefPubMedGoogle Scholar
  • 5 Wesselschmidt R, Likert K, Girard T, Wun TC, Broze Jr GJ. Tissue factor pathway inhibitor: the carboxy-terminus is required for optimal inhibition of factor Xa. Blood 1992; 79 (08) 2004-10.
    PubMedGoogle Scholar
  • 6 Wesselschmidt R, Likert K, Huang Z, MacPhail L, Broze Jr GJ. Structural requirements for tissue factor pathway inhibitor interactions with factor Xa and heparin. Blood Coagul Fibrinolysis 1993; 4 (05) 661-9.
    CrossrefPubMedGoogle Scholar
  • 7 Lindhout T, Willems G, Blezer R, Hemker HC. Kinetics of the inhibition of human factor Xa by full-length and truncated recombinant tissue factor pathway inhibitor. Biochem J 1994; 297 (Pt 1) 131-6.
    CrossrefPubMedGoogle Scholar
  • 8 Warshawsky I, Bu G, Mast A, Saffitz JE, Broze Jr. GJ, Schwartz AL. The carboxy terminus of tissue factor pathway inhibitor is required for interacting with hepatoma cells in vitro and in vivo. J Clin Invest 1995; 95 (04) 1773-81.
    CrossrefPubMedGoogle Scholar
  • 9 Mast AE, Stadanlick JE, Lockett JM, Dietzen DJ, Hasty KA, Hall CL. Tissue factor pathway inhibitor binds to platelet thrombospondin-1. J Biol Chem 2000; 275 (41) 31715-21.
    CrossrefPubMedGoogle Scholar
  • 10 Novotny WF, Brown SG, Miletich JP, Rader DJ, Broze Jr GJ. Plasma antigen levels of the lipoprotein-associated coagulation inhibitor in patient samples. Blood 1991; 78 (02) 387-93.
    PubMedGoogle Scholar
  • 11 Osterud B, Bajaj MS, Bajaj SP. Sites of tissue factor pathway inhibitor (TFPI) and tissue factor expression under physiologic and pathologic conditions. On behalf of the Subcommittee on Tissue Factor Pathway Inhibitor (TFPI) of the Scientific and Standardization Committee of the ISTH. Thromb Haemost 1995; 73 (05) 873-5.
    Artikel in Thieme ConnectPubMedGoogle Scholar
  • 12 Werling RW, Zacharski LR, Kisiel W, Bajaj SP, Memoli VA, Rousseau SM. Distribution of tissue factor pathway inhibitor in normal and malignant human tissues. Thromb Haemost 1993; 69 (04) 366-9.
    Artikel in Thieme ConnectPubMedGoogle Scholar
  • 13 Sandset PM, Abildgaard U, Larsen ML. Heparin induces release of extrinsic coagulation pathway inhibitor (EPI). Thromb Res 1988; 50 (06) 803-13.
    CrossrefPubMedGoogle Scholar
  • 14 Lupu C, Poulsen E, Roquefeuil S, West-muckett AD, Kakkar VV, Lupu F. Cellular effects of heparin on the production and release of tissue factor pathway inhibitor in human endothelial cells in culture. Arterioscler Thromb Vasc Biol 1999; 19 (09) 2251-62.
    CrossrefPubMedGoogle Scholar
  • 15 Lupu C, Goodwin CA, Westmuckett AD, Emeis JJ, Scully MF, Kakkar VV. et al. Tissue factor pathway inhibitor in endothelial cells colocalizes with glycolipid microdomains/ caveolae. Regulatory mechanism(s) of the anticoagulant properties of the endothelium. Arterioscler Thromb Vasc Biol 1997; 17 (11) 2964-74.
    CrossrefPubMedGoogle Scholar
  • 16 Ott I, Miyagi Y, Miyazaki K, Heeb MJ, Mueller BM, Rao LV. et al. Reversible regulation of tissue factor-induced coagulation by glycosyl phosphatidylinositol-anchored tissue factor pathway inhibitor. Arterioscler Thromb Vasc Biol 2000; 20 (03) 874-82.
    CrossrefPubMedGoogle Scholar
  • 17 Mast AE, Higuchi DA, Huang ZF, Warshaw-sky I, Schwartz AL, Broze Jr GJ. Glypican-3 is a binding protein on the HepG2 cell surface for tissue factor pathway inhibitor. Biochem J 1997; 327 (Pt 2) 577-83.
    CrossrefPubMedGoogle Scholar
  • 18 Cinek T, Horejsi V. The nature of large noncovalent complexes containing glycosyl-phosphatidylinositol-anchored membrane glyco-proteins and protein tyrosine kinases. J Immunol 1992; 149 (07) 2262-70.
    PubMedGoogle Scholar
  • 19 Brown DA, Rose JK. Sorting of GPI-anchored proteins to glycolipid-enriched membrane subdomains during transport to the apical cell surface. Cell 1992; 68 (03) 533-44.
    CrossrefPubMedGoogle Scholar
  • 20 Rothberg KG, Heuser JE, Donzell WC, Ying YS, Glenney JR, Anderson RG. Caveolin, a protein component of caveolae membrane coats. Cell 1992; 68 (04) 673-82.
    CrossrefPubMedGoogle Scholar
  • 21 Yamada E. The fine structure of the gall bladder epithelium of the mouse. J Biophys Biochem Cytol 1955; 445-58.
    PubMedGoogle Scholar
  • 22 Simons K, Ikonen E. Functional rafts in cell membranes. Nature 1997; 387 6633 569-72.
    CrossrefPubMedGoogle Scholar
  • 23 Fra AM, Williamson E, Simons K, Parton RG. De novo formation of caveolae in lymphocytes by expression of VIP21-caveolin. Proc Natl Acad Sci USA 1995; 92 (19) 8655-9.
    CrossrefPubMedGoogle Scholar
  • 24 Fra AM, Williamson E, Simons K, Parton RG. Detergent-insoluble glycolipid microdomains in lymphocytes in the absence of caveolae. J Biol Chem 1994; 269 (49) 30745-8.
    PubMedGoogle Scholar
  • 25 Anderson RG, Kamen BA, Rothberg KG, Lacey SW. Potocytosis: sequestration and transport of small molecules by caveolae. Science 1992; 255 5043 410-11.
    CrossrefPubMedGoogle Scholar
  • 26 Smart EJ, Ying YS, Conrad PA, Anderson RG. Caveolin moves from caveolae to the Golgi apparatus in response to cholesterol oxidation. J Cell Biol 1994; 127 (05) 1185-97.
    CrossrefPubMedGoogle Scholar
  • 27 Smart EJ, Ying Y, Donzell WC, Anderson RG. A role for caveolin in transport of cholesterol from endoplasmic reticulum to plasma membrane. J Biol Chem 1996; 271 (46) 29427-35.
    CrossrefPubMedGoogle Scholar
  • 28 Nieva JL, Bron R, Corver J, Wilschut J. Membrane fusion of Semliki Forest virus requires sphingolipids in the target membrane. EMBO J 1994; 13 (12) 2797-804.
    PubMedGoogle Scholar
  • 29 Phalen T, Kielian M. Cholesterol is required for infection by Semliki Forest virus. J Cell Biol 1991; 112 (04) 615-23.
    CrossrefPubMedGoogle Scholar
  • 30 Stang E, Kartenbeck J, Parton RG. Major his-tocompatibility complex class I molecules mediate association of SV40 with caveolae. Mol Biol Cell 1997; 8 (01) 47-57.
    CrossrefPubMedGoogle Scholar
  • 31 Baorto DM, Gao Z, Malaviya R, Dustin ML, van der MA, Lublin DM. et al. Survival of FimH-expressing enterobacteria in macrophages relies on glycolipid traffic. Nature 1997; 389 6651 636-9.
    CrossrefPubMedGoogle Scholar
  • 32 Field KA, Holowka D, Baird B. Fc epsilon RI-mediated recruitment of p53/56lyn to detergent-resistant membrane domains accompanies cellular signaling. Proc Natl Acad Sci USA 1995; 92 (20) 9201-5.
    CrossrefPubMedGoogle Scholar
  • 33 Chun M, Liyanage UK, Lisanti MP, Lodish HF. Signal transduction of a G protein-coupled receptor in caveolae: colocalization of endothelin and its receptor with caveolin. Proc Natl Acad Sci USA 1994; 91 (24) 11728-32.
    CrossrefPubMedGoogle Scholar
  • 34 Sevinsky JR, Rao LV, Ruf W. Ligand-induced protease receptor translocation into caveolae: a mechanism for regulating cell surface proteolysis of the tissue factor-dependent coagulation pathway. J Cell Biol 1996; 133 (02) 293-304.
    CrossrefPubMedGoogle Scholar
  • 35 Medof ME, Lublin DM, Holers VM, Ayers DJ, Getty RR, Leykam JF. et al. Cloning and characterization of cDNAs encoding the complete sequence of decay-accelerating factor of human complement. Proc Natl Acad Sci USA 1987; 84 (07) 2007-11.
    CrossrefPubMedGoogle Scholar
  • 36 Hansbrough JR, Lublin DM, Roth KA, Birkenmeier EA, Gordon JI. Expression of a liver fatty acid binding protein/human decay-accelerating factor/HLA-B44 chimeric gene in transgenic mice. Am J Physiol 1991; 260 (6 Pt 1) G929-G939.
    PubMedGoogle Scholar
  • 37 Edidin M, Kuo SC, Sheetz MP. Lateral movements of membrane glycoproteins restricted by dynamic cytoplasmic barriers. Science 1991; 254 5036 1379-82.
    CrossrefPubMedGoogle Scholar
  • 38 Lublin DM, Coyne KE. Phospholipid-anchored and transmembrane versions of either decay-accelerating factor or membrane cofactor protein show equal efficiency in protection from complement-mediated cell damage. J Exp Med 1991; 174 (01) 35-44.
    CrossrefPubMedGoogle Scholar
  • 39 Zhang F, Crise B, Su B, Hou Y, Rose JK, Bothwell A. et al. The lateral mobility of some membrane proteins is determined by their ectodomains. Biophys J 1992; 62 (01) 92-4.
    CrossrefPubMedGoogle Scholar
  • 40 Zhang F, Crise B, Su B, Hou Y, Rose JK, Bothwell A. et al. Lateral diffusion of membrane-spanning and glycosylphosphatidyl-inositol-linked proteins: toward establishing rules governing the lateral mobility of membrane proteins. J Cell Biol 1991; 115 (01) 75-84.
    CrossrefPubMedGoogle Scholar
  • 41 Feng X, Gaeta ML, Madge LA, Yang JH, Bradley JR, Pober JS. Caveolin-1 associates with TRAF2 to form a complex that is recruited to tumor necrosis factor receptors. J Biol Chem 2001; 276 (11) 8341-9.
    CrossrefPubMedGoogle Scholar
  • 42 Xu J, Qu D, Esmon NL, Esmon CT. Metallo-proteolytic release of endothelial cell protein C receptor. J Biol Chem 2000; 275 (08) 6038-44.
    CrossrefPubMedGoogle Scholar
  • 43 Le DT, Rapaport SI, Rao LV. Relations between factor VIIa binding and expression of factor VIIa/tissue factor catalytic activity on cell surfaces. J Biol Chem 1992; 267 (22) 15447-54.
    PubMedGoogle Scholar
  • 44 Wolberg AS, Monroe DM, Roberts HR, Hoffman MR. Tissue factor de-encryption: ionophore treatment induces changes in tissue factor activity by phosphatidylserine-dependent and -independent mechanisms. Blood Coagul Fibrinolysis 1999; 10 (04) 201-10.
    CrossrefPubMedGoogle Scholar