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. 1996 Feb 1;132(3):487–497. doi: 10.1083/jcb.132.3.487

Heparan sulfate expression in polarized epithelial cells: the apical sorting of glypican (GPI-anchored proteoglycan) is inversely related to its heparan sulfate content

PMCID: PMC2120721  PMID: 8636224

Abstract

Several processes that occur in the luminal compartments of the tissues are modulated by heparin-like polysaccharides. To identify proteins responsible for the expression of heparan sulfate at the apex of polarized cells, we investigated the polarity of the expression of the cell surface heparan sulfate proteoglycans in CaCo-2 cells. Domain- specific biotinylation of the apical and basolateral membranes of these cells identified glypican, a GPI-linked heparan sulfate proteoglycan, as the major source of apical heparan sulfate. Yet, most of this proteoglycan was expressed at the basolateral surface, an unexpected finding for a glypiated protein. Metabolic labeling and chase experiments indicated that sorting mechanisms, rather than differential turnover, accounted for this bipolar expression of glypican. Chlorate treatment did not affect the polarity of the expression of glypican in CaCo-2 cells, and transfectant MDCK cells expressed wild-type glypican and a syndecan-4/glypican chimera also in an essentially unpolarized fashion. Yet, complete removal of the heparan sulfate glycanation sites from the glypican core protein resulted in the nearly exclusive apical targeting of glypican in the transfectants, whereas two- and one-chain mutant forms had intermediate distributions. These results indicate that glypican accounts for the expression of apical heparan sulfate, but that glycanation of the core protein antagonizes the activity of the apical sorting signal conveyed by the GPI anchor of this proteoglycan. A possible implication of these findings is that heparan sulfate glycanation may be a determinant of the subcellular expression of glypican. Alternatively, inverse glycanation-apical sorting relationships in glypican may insure near constant deliveries of HS to the apical compartment, or "active" GPI-mediated entry of heparan sulfate into apical membrane compartments may require the overriding of this antagonizing effect of the heparan sulfate chains.

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Selected References

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  1. Andres J. L., Stanley K., Cheifetz S., Massagué J. Membrane-anchored and soluble forms of betaglycan, a polymorphic proteoglycan that binds transforming growth factor-beta. J Cell Biol. 1989 Dec;109(6 Pt 1):3137–3145. doi: 10.1083/jcb.109.6.3137. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Bernfield M., Kokenyesi R., Kato M., Hinkes M. T., Spring J., Gallo R. L., Lose E. J. Biology of the syndecans: a family of transmembrane heparan sulfate proteoglycans. Annu Rev Cell Biol. 1992;8:365–393. doi: 10.1146/annurev.cb.08.110192.002053. [DOI] [PubMed] [Google Scholar]
  3. Bomsel M., Mostov K. Sorting of plasma membrane proteins in epithelial cells. Curr Opin Cell Biol. 1991 Aug;3(4):647–653. doi: 10.1016/0955-0674(91)90036-x. [DOI] [PubMed] [Google Scholar]
  4. Bourin M. C., Lindahl U. Glycosaminoglycans and the regulation of blood coagulation. Biochem J. 1993 Jan 15;289(Pt 2):313–330. doi: 10.1042/bj2890313. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Brown D. A., Rose J. K. Sorting of GPI-anchored proteins to glycolipid-enriched membrane subdomains during transport to the apical cell surface. Cell. 1992 Feb 7;68(3):533–544. doi: 10.1016/0092-8674(92)90189-j. [DOI] [PubMed] [Google Scholar]
  6. David G. Integral membrane heparan sulfate proteoglycans. FASEB J. 1993 Aug;7(11):1023–1030. doi: 10.1096/fasebj.7.11.8370471. [DOI] [PubMed] [Google Scholar]
  7. David G., Lories V., Decock B., Marynen P., Cassiman J. J., 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. doi: 10.1083/jcb.111.6.3165. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. David G., van der Schueren B., Marynen P., Cassiman J. J., van den Berghe H. Molecular cloning of amphiglycan, a novel integral membrane heparan sulfate proteoglycan expressed by epithelial and fibroblastic cells. J Cell Biol. 1992 Aug;118(4):961–969. doi: 10.1083/jcb.118.4.961. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Filmus J., Church J. G., Buick R. N. Isolation of a cDNA corresponding to a developmentally regulated transcript in rat intestine. Mol Cell Biol. 1988 Oct;8(10):4243–4249. doi: 10.1128/mcb.8.10.4243. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Garcia M., Seigner C., Bastid C., Choux R., Payan M. J., Reggio H. Carcinoembryonic antigen has a different molecular weight in normal colon and in cancer cells due to N-glycosylation differences. Cancer Res. 1991 Oct 15;51(20):5679–5686. [PubMed] [Google Scholar]
  11. Gitay-Goren H., Soker S., Vlodavsky I., Neufeld G. The binding of vascular endothelial growth factor to its receptors is dependent on cell surface-associated heparin-like molecules. J Biol Chem. 1992 Mar 25;267(9):6093–6098. [PubMed] [Google Scholar]
  12. Hooper N. M. More than just a membrane anchor. Curr Biol. 1992 Nov;2(11):617–619. doi: 10.1016/0960-9822(92)90183-b. [DOI] [PubMed] [Google Scholar]
  13. Hooper N. M., Turner A. J. Ectoenzymes of the kidney microvillar membrane. Differential solubilization by detergents can predict a glycosyl-phosphatidylinositol membrane anchor. Biochem J. 1988 Mar 15;250(3):865–869. doi: 10.1042/bj2500865. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Hopkins C. R. Polarity signals. Cell. 1991 Sep 6;66(5):827–829. doi: 10.1016/0092-8674(91)90427-z. [DOI] [PubMed] [Google Scholar]
  15. Karthikeyan L., Flad M., Engel M., Meyer-Puttlitz B., Margolis R. U., Margolis R. K. Immunocytochemical and in situ hybridization studies of the heparan sulfate proteoglycan, glypican, in nervous tissue. J Cell Sci. 1994 Nov;107(Pt 11):3213–3222. doi: 10.1242/jcs.107.11.3213. [DOI] [PubMed] [Google Scholar]
  16. Kojima T., Leone C. W., Marchildon G. A., Marcum J. A., Rosenberg R. D. Isolation and characterization of heparan sulfate proteoglycans produced by cloned rat microvascular endothelial cells. J Biol Chem. 1992 Mar 5;267(7):4859–4869. [PubMed] [Google Scholar]
  17. Le Bivic A., Quaroni A., Nichols B., Rodriguez-Boulan E. Biogenetic pathways of plasma membrane proteins in Caco-2, a human intestinal epithelial cell line. J Cell Biol. 1990 Oct;111(4):1351–1361. doi: 10.1083/jcb.111.4.1351. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Le Bivic A., Real F. X., Rodriguez-Boulan E. Vectorial targeting of apical and basolateral plasma membrane proteins in a human adenocarcinoma epithelial cell line. Proc Natl Acad Sci U S A. 1989 Dec;86(23):9313–9317. doi: 10.1073/pnas.86.23.9313. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Lisanti M. P., Le Bivic A., Saltiel A. R., Rodriguez-Boulan E. Preferred apical distribution of glycosyl-phosphatidylinositol (GPI) anchored proteins: a highly conserved feature of the polarized epithelial cell phenotype. J Membr Biol. 1990 Feb;113(2):155–167. doi: 10.1007/BF01872889. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Lisanti M. P., Rodriguez-Boulan E. Glycophospholipid membrane anchoring provides clues to the mechanism of protein sorting in polarized epithelial cells. Trends Biochem Sci. 1990 Mar;15(3):113–118. doi: 10.1016/0968-0004(90)90195-h. [DOI] [PubMed] [Google Scholar]
  21. Lories V., Cassiman J. J., Van den Berghe H., David G. Differential expression of cell surface heparan sulfate proteoglycans in human mammary epithelial cells and lung fibroblasts. J Biol Chem. 1992 Jan 15;267(2):1116–1122. [PubMed] [Google Scholar]
  22. Lories V., Cassiman J. J., Van den Berghe H., David G. Multiple distinct membrane heparan sulfate proteoglycans in human lung fibroblasts. J Biol Chem. 1989 Apr 25;264(12):7009–7016. [PubMed] [Google Scholar]
  23. Lories V., De Boeck H., David G., Cassiman J. J., Van den Berghe H. Heparan sulfate proteoglycans of human lung fibroblasts. Structural heterogeneity of the core proteins of the hydrophobic cell-associated forms. J Biol Chem. 1987 Jan 15;262(2):854–859. [PubMed] [Google Scholar]
  24. Mertens G., Cassiman J. J., Van den Berghe H., Vermylen J., David G. Cell surface heparan sulfate proteoglycans from human vascular endothelial cells. Core protein characterization and antithrombin III binding properties. J Biol Chem. 1992 Oct 5;267(28):20435–20443. [PubMed] [Google Scholar]
  25. Miettinen H. M., Edwards S. N., Jalkanen M. Analysis of transport and targeting of syndecan-1: effect of cytoplasmic tail deletions. Mol Biol Cell. 1994 Dec;5(12):1325–1339. doi: 10.1091/mbc.5.12.1325. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Mostov K., Apodaca G., Aroeti B., Okamoto C. Plasma membrane protein sorting in polarized epithelial cells. J Cell Biol. 1992 Feb;116(3):577–583. doi: 10.1083/jcb.116.3.577. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Rapraeger A. C., Krufka A., Olwin B. B. Requirement of heparan sulfate for bFGF-mediated fibroblast growth and myoblast differentiation. Science. 1991 Jun 21;252(5013):1705–1708. doi: 10.1126/science.1646484. [DOI] [PubMed] [Google Scholar]
  28. Rapraeger A., Jalkanen M., Bernfield M. Cell surface proteoglycan associates with the cytoskeleton at the basolateral cell surface of mouse mammary epithelial cells. J Cell Biol. 1986 Dec;103(6 Pt 2):2683–2696. doi: 10.1083/jcb.103.6.2683. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Rapraeger A. Transforming growth factor (type beta) promotes the addition of chondroitin sulfate chains to the cell surface proteoglycan (syndecan) of mouse mammary epithelia. J Cell Biol. 1989 Nov;109(5):2509–2518. doi: 10.1083/jcb.109.5.2509. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Sargiacomo M., Lisanti M., Graeve L., Le Bivic A., Rodriguez-Boulan E. Integral and peripheral protein composition of the apical and basolateral membrane domains in MDCK cells. J Membr Biol. 1989 Mar;107(3):277–286. doi: 10.1007/BF01871942. [DOI] [PubMed] [Google Scholar]
  31. Saxena U., Klein M. G., Goldberg I. J. Identification and characterization of the endothelial cell surface lipoprotein lipase receptor. J Biol Chem. 1991 Sep 15;266(26):17516–17521. [PubMed] [Google Scholar]
  32. Saxena U., Klein M. G., Goldberg I. J. Transport of lipoprotein lipase across endothelial cells. Proc Natl Acad Sci U S A. 1991 Mar 15;88(6):2254–2258. doi: 10.1073/pnas.88.6.2254. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Schell M. J., Maurice M., Stieger B., Hubbard A. L. 5'nucleotidase is sorted to the apical domain of hepatocytes via an indirect route. J Cell Biol. 1992 Dec;119(5):1173–1182. doi: 10.1083/jcb.119.5.1173. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Simons K., Wandinger-Ness A. Polarized sorting in epithelia. Cell. 1990 Jul 27;62(2):207–210. doi: 10.1016/0092-8674(90)90357-k. [DOI] [PubMed] [Google Scholar]
  35. Simons K., van Meer G. Lipid sorting in epithelial cells. Biochemistry. 1988 Aug 23;27(17):6197–6202. doi: 10.1021/bi00417a001. [DOI] [PubMed] [Google Scholar]
  36. Spivak-Kroizman T., Lemmon M. A., Dikic I., Ladbury J. E., Pinchasi D., Huang J., Jaye M., Crumley G., Schlessinger J., Lax I. Heparin-induced oligomerization of FGF molecules is responsible for FGF receptor dimerization, activation, and cell proliferation. Cell. 1994 Dec 16;79(6):1015–1024. doi: 10.1016/0092-8674(94)90032-9. [DOI] [PubMed] [Google Scholar]
  37. Stipp C. S., Litwack E. D., Lander A. D. 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. doi: 10.1083/jcb.124.1.149. [DOI] [PMC free article] [PubMed] [Google Scholar]
  38. Tanaka Y., Adams D. H., Hubscher S., Hirano H., Siebenlist U., Shaw S. T-cell adhesion induced by proteoglycan-immobilized cytokine MIP-1 beta. Nature. 1993 Jan 7;361(6407):79–82. doi: 10.1038/361079a0. [DOI] [PubMed] [Google Scholar]
  39. 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. doi: 10.1083/jcb.130.5.1207. [DOI] [PMC free article] [PubMed] [Google Scholar]
  40. Yanagishita M. Glycosylphosphatidylinositol-anchored and core protein-intercalated heparan sulfate proteoglycans in rat ovarian granulosa cells have distinct secretory, endocytotic, and intracellular degradative pathways. J Biol Chem. 1992 May 15;267(14):9505–9511. [PubMed] [Google Scholar]
  41. Yayon A., Klagsbrun M., Esko J. D., Leder P., Ornitz D. M. Cell surface, heparin-like molecules are required for binding of basic fibroblast growth factor to its high affinity receptor. Cell. 1991 Feb 22;64(4):841–848. doi: 10.1016/0092-8674(91)90512-w. [DOI] [PubMed] [Google Scholar]
  42. Zurzolo C., Rodriguez-Boulan E. Delivery of Na+,K(+)-ATPase in polarized epithelial cells. Science. 1993 Apr 23;260(5107):550–556. doi: 10.1126/science.8386394. [DOI] [PubMed] [Google Scholar]
  43. Zurzolo C., van't Hof W., van Meer G., Rodriguez-Boulan E. VIP21/caveolin, glycosphingolipid clusters and the sorting of glycosylphosphatidylinositol-anchored proteins in epithelial cells. EMBO J. 1994 Jan 1;13(1):42–53. doi: 10.1002/j.1460-2075.1994.tb06233.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  44. de Boeck H., Lories V., David G., Cassiman J. J., van den Berghe H. Identification of a 64 kDa heparan sulphate proteoglycan core protein from human lung fibroblast plasma membranes with a monoclonal antibody. Biochem J. 1987 Nov 1;247(3):765–771. doi: 10.1042/bj2470765. [DOI] [PMC free article] [PubMed] [Google Scholar]

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