• We are sorry, but NCBI web applications do not support your browser and may not function properly. More information
Logo of biochemjBJ Latest papers and much more!
Biochem J. Oct 15, 1997; 327(Pt 2): 577–583.
PMCID: PMC1218832

Glypican-3 is a binding protein on the HepG2 cell surface for tissue factor pathway inhibitor.

Abstract

Tissue factor pathway inhibitor (TFPI) is a primary regulator of the initiation of blood coagulation. TFPI is internalized and degraded by HepG2 cells through the low-density-lipoprotein receptor-related protein (LRP) but also binds another molecule present on the cell surface at approx. 10-fold the abundance of LRP [Warshawsky, Broze and Schwartz (1994) Proc. Natl. Acad. Sci. U.S.A. 91, 6664-6668]. When HepG2 cells are washed with heparin or dextran sulphate, a substance that binds TFPI is removed from the cell surface and can be detected in a slot-blot assay. Preincubation with trypsin destroys the reactivity of the TFPI-binding component in the slot-blot assay, suggesting that it is a protein. In addition, when the sulphation of glycosaminoglycans (GAGs) is prevented by growing the HepG2 cells in the presence of 30 mM sodium chlorate, TFPI binding is unaffected, whereas the binding of bovine lipoprotein lipase, a protein known to associate with cell-surface GAGs, falls to 50% of control levels. Dextran sulphate washes of HepG2 cells grown in sodium chlorate have an equal reactivity in slot-blot experiments to that of non-treated cells, suggesting that GAGs are not totally responsible for the binding activity observed. By using the slot blot to follow binding activity and conventional protein purification techniques, a protein species that migrates at 40 kDa after reduction was identified in the HepG2 cell wash. The binding of this protein to TFPI was confirmed with immobilized TFPI. Amino acid sequence analysis identified this protein species as a proteolytic fragment of glypican-3 (also called OCI-5), a member of the glypican family of glycosylphosphatidylinositol-anchored proteoglycans.

Full Text

The Full Text of this article is available as a PDF (377K).

Selected References

These references are in PubMed. This may not be the complete list of references from this article.
  • Girard TJ, Warren LA, Novotny WF, Likert KM, Brown SG, Miletich JP, Broze GJ., Jr Functional significance of the Kunitz-type inhibitory domains of lipoprotein-associated coagulation inhibitor. Nature. 1989 Apr 6;338(6215):518–520. [PubMed]
  • Wesselschmidt R, Likert K, Girard T, Wun TC, Broze GJ., Jr Tissue factor pathway inhibitor: the carboxy-terminus is required for optimal inhibition of factor Xa. Blood. 1992 Apr 15;79(8):2004–2010. [PubMed]
  • 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 Jan 1;297(Pt 1):131–136. [PMC free article] [PubMed]
  • Wesselschmidt R, Likert K, Huang Z, MacPhail L, Broze GJ., Jr Structural requirements for tissue factor pathway inhibitor interactions with factor Xa and heparin. Blood Coagul Fibrinolysis. 1993 Oct;4(5):661–669. [PubMed]
  • Broze GJ, Jr, Lange GW, Duffin KL, MacPhail L. Heterogeneity of plasma tissue factor pathway inhibitor. Blood Coagul Fibrinolysis. 1994 Aug;5(4):551–559. [PubMed]
  • Sandset PM, Abildgaard U, Larsen ML. Heparin induces release of extrinsic coagulation pathway inhibitor (EPI). Thromb Res. 1988 Jun 15;50(6):803–813. [PubMed]
  • Novotny WF, Palmier M, Wun TC, Broze GJ, Jr, Miletich JP. Purification and properties of heparin-releasable lipoprotein-associated coagulation inhibitor. Blood. 1991 Jul 15;78(2):394–400. [PubMed]
  • Novotny WF, Brown SG, Miletich JP, Rader DJ, Broze GJ., Jr Plasma antigen levels of the lipoprotein-associated coagulation inhibitor in patient samples. Blood. 1991 Jul 15;78(2):387–393. [PubMed]
  • Palmier MO, Hall LJ, Reisch CM, Baldwin MK, Wilson AG, Wun TC. Clearance of recombinant tissue factor pathway inhibitor (TFPI) in rabbits. Thromb Haemost. 1992 Jul 6;68(1):33–36. [PubMed]
  • Bregengaard C, Nordfang O, Ostergaard P, Petersen JG, Meyn G, Diness V, Svendsen O, Hedner U. Pharmacokinetics of full length and two-domain tissue factor pathway inhibitor in combination with heparin in rabbits. Thromb Haemost. 1993 Sep 1;70(3):454–457. [PubMed]
  • Warshawsky I, Bu G, Mast A, Saffitz JE, Broze GJ, Jr, 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 Apr;95(4):1773–1781. [PMC free article] [PubMed]
  • Warshawsky I, Broze GJ, Jr, Schwartz AL. The low density lipoprotein receptor-related protein mediates the cellular degradation of tissue factor pathway inhibitor. Proc Natl Acad Sci U S A. 1994 Jul 5;91(14):6664–6668. [PMC free article] [PubMed]
  • Warshawsky I, Bu G, Schwartz AL. Identification of domains on the 39-kDa protein that inhibit the binding of ligands to the low density lipoprotein receptor-related protein. J Biol Chem. 1993 Oct 15;268(29):22046–22054. [PubMed]
  • Broze GJ, Jr, Miletich JP. Isolation of the tissue factor inhibitor produced by HepG2 hepatoma cells. Proc Natl Acad Sci U S A. 1987 Apr;84(7):1886–1890. [PMC free article] [PubMed]
  • David G, Lories V, Decock B, Marynen P, Cassiman JJ, Van den Berghe H. Molecular cloning of a phosphatidylinositol-anchored membrane heparan sulfate proteoglycan from human lung fibroblasts. J Cell Biol. 1990 Dec;111(6 Pt 2):3165–3176. [PMC free article] [PubMed]
  • Watanabe K, Yamada H, Yamaguchi Y. K-glypican: a novel GPI-anchored heparan sulfate proteoglycan that is highly expressed in developing brain and kidney. J Cell Biol. 1995 Sep;130(5):1207–1218. [PMC free article] [PubMed]
  • Pilia G, Hughes-Benzie RM, MacKenzie A, Baybayan P, Chen EY, Huber R, Neri G, Cao A, Forabosco A, Schlessinger D. Mutations in GPC3, a glypican gene, cause the Simpson-Golabi-Behmel overgrowth syndrome. Nat Genet. 1996 Mar;12(3):241–247. [PubMed]
  • Broze GJ, Jr, Warren LA, Girard JJ, Miletich JP. Isolation of the lipoprotein associated coagulation inhibitor produced by HepG2 (human hepatoma) cells using bovine factor Xa affinity chromatography. Thromb Res. 1987 Oct 15;48(2):253–259. [PubMed]
  • Zhang L, David G, Esko JD. Repetitive Ser-Gly sequences enhance heparan sulfate assembly in proteoglycans. J Biol Chem. 1995 Nov 10;270(45):27127–27135. [PubMed]
  • Lindahl AK, Sandset PM, Abildgaard U. The present status of tissue factor pathway inhibitor. Blood Coagul Fibrinolysis. 1992 Aug;3(4):439–449. [PubMed]
  • Baeuerle PA, Huttner WB. Chlorate--a potent inhibitor of protein sulfation in intact cells. Biochem Biophys Res Commun. 1986 Dec 15;141(2):870–877. [PubMed]
  • Cisar LA, Hoogewerf AJ, Cupp M, Rapport CA, Bensadoun A. Secretion and degradation of lipoprotein lipase in cultured adipocytes. Binding of lipoprotein lipase to membrane heparan sulfate proteoglycans is necessary for degradation. J Biol Chem. 1989 Jan 25;264(3):1767–1774. [PubMed]
  • Hoogewerf AJ, Cisar LA, Evans DC, Bensadoun A. Effect of chlorate on the sulfation of lipoprotein lipase and heparan sulfate proteoglycans. Sulfation of heparan sulfate proteoglycans affects lipoprotein lipase degradation. J Biol Chem. 1991 Sep 5;266(25):16564–16571. [PubMed]
  • Filmus J, Church JG, Buick RN. Isolation of a cDNA corresponding to a developmentally regulated transcript in rat intestine. Mol Cell Biol. 1988 Oct;8(10):4243–4249. [PMC free article] [PubMed]
  • Stipp CS, Litwack ED, Lander AD. Cerebroglycan: an integral membrane heparan sulfate proteoglycan that is unique to the developing nervous system and expressed specifically during neuronal differentiation. J Cell Biol. 1994 Jan;124(1-2):149–160. [PMC free article] [PubMed]
  • Filmus J, Shi W, Wong ZM, Wong MJ. Identification of a new membrane-bound heparan sulphate proteoglycan. Biochem J. 1995 Oct 15;311(Pt 2):561–565. [PMC free article] [PubMed]
  • Kjellén L, Oldberg A, Hök M. Cell-surface heparan sulfate. Mechanisms of proteoglycan-cell association. J Biol Chem. 1980 Nov 10;255(21):10407–10413. [PubMed]
  • Ishihara M, Fedarko NS, Conrad HE. Involvement of phosphatidylinositol and insulin in the coordinate regulation of proteoheparan sulfate metabolism and hepatocyte growth. J Biol Chem. 1987 Apr 5;262(10):4708–4716. [PubMed]
  • 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 Apr;133(2):293–304. [PMC free article] [PubMed]
  • Iversen N, Sandset PM, Abildgaard U, Torjesen PA. Binding of tissue factor pathway inhibitor to cultured endothelial cells-influence of glycosaminoglycans. Thromb Res. 1996 Nov 15;84(4):267–278. [PubMed]
  • Ho G, Toomey JR, Broze GJ, Jr, Schwartz AL. Receptor-mediated endocytosis of coagulation factor Xa requires cell surface-bound tissue factor pathway inhibitor. J Biol Chem. 1996 Apr 19;271(16):9497–9502. [PubMed]

Articles from Biochemical Journal are provided here courtesy of The Biochemical Society

Formats:

Related citations in PubMed

See reviews...See all...

Cited by other articles in PMC

See all...

Links

  • Cited in Books
    Cited in Books
    PubMed Central articles cited in books
  • Compound
    Compound
    PubChem Compound links
  • MedGen
    MedGen
    Related information in MedGen
  • PubMed
    PubMed
    PubMed citations for these articles
  • Substance
    Substance
    PubChem Substance links

Recent Activity

Your browsing activity is empty.

Activity recording is turned off.

Turn recording back on

See more...