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. 1994 Feb;5(2):183–192. doi: 10.1091/mbc.5.2.183

Syndecan 4 heparan sulfate proteoglycan is a selectively enriched and widespread focal adhesion component.

A Woods 1, J R Couchman 1
PMCID: PMC301024  PMID: 8019004

Abstract

Focal adhesion formation in fibroblasts results from complex transmembrane signaling processes initiated by extracellular matrix molecules. Although a role for integrins with attendant tyrosine kinases has been established, there is evidence that cell surface heparan sulfate proteoglycans (HSPGs) are also involved with an associated role of protein kinase C. The identity of the proteoglycan has remained elusive, but we now report that syndecan 4 (ryudocan/amphiglycan) is present in focal adhesions of a number of cell types. Affinity-purified antibodies raised against a unique portion of the cytoplasmic domain of syndecan 4 core protein recognized an HSPG of similar characteristics to those of syndecan 4. These antibodies stained focal adhesions only after cell permeabilization and recognized differing mammalian species. Syndecan 4 was associated with focal adhesions that contained either beta 1 or beta 3 integrin subunits and those that formed on substrates of fibronectin, laminin, vitronectin, or type I collagen. No focal adhesions were found that were vinculin-containing but lacked syndecan 4. In contrast, syndecan 2, whose cytoplasmic domain is closely homologous to syndecan 4, does not appear to be a focal adhesion component. Thus, syndecan 4 represents a new transmembrane focal adhesion component, probably involved in their assembly.

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Badley R. A., Woods A., Rees D. A. Cooperativity of concanavalin A patching and its influence on cytoskeleton changes in fibroblast rounding and detachment. J Cell Sci. 1981 Feb;47:349–363. doi: 10.1242/jcs.47.1.349. [DOI] [PubMed] [Google Scholar]
  2. Balzac F., Belkin A. M., Koteliansky V. E., Balabanov Y. V., Altruda F., Silengo L., Tarone G. Expression and functional analysis of a cytoplasmic domain variant of the beta 1 integrin subunit. J Cell Biol. 1993 Apr;121(1):171–178. doi: 10.1083/jcb.121.1.171. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. 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]
  4. Burridge K., Fath K., Kelly T., Nuckolls G., Turner C. Focal adhesions: transmembrane junctions between the extracellular matrix and the cytoskeleton. Annu Rev Cell Biol. 1988;4:487–525. doi: 10.1146/annurev.cb.04.110188.002415. [DOI] [PubMed] [Google Scholar]
  5. Burridge K., Turner C. E., Romer L. H. Tyrosine phosphorylation of paxillin and pp125FAK accompanies cell adhesion to extracellular matrix: a role in cytoskeletal assembly. J Cell Biol. 1992 Nov;119(4):893–903. doi: 10.1083/jcb.119.4.893. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Carey D. J., Evans D. M., Stahl R. C., Asundi V. K., Conner K. J., Garbes P., Cizmeci-Smith G. Molecular cloning and characterization of N-syndecan, a novel transmembrane heparan sulfate proteoglycan. J Cell Biol. 1992 Apr;117(1):191–201. doi: 10.1083/jcb.117.1.191. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Couchman J. R., Austria R., Woods A., Hughes R. C. Adhesion defective BHK cell mutant has cell surface heparan sulfate proteoglycan of altered properties. J Cell Physiol. 1988 Aug;136(2):226–236. doi: 10.1002/jcp.1041360204. [DOI] [PubMed] [Google Scholar]
  8. Couchman J. R. Heterogeneous distribution of a basement membrane heparan sulfate proteoglycan in rat tissues. J Cell Biol. 1987 Oct;105(4):1901–1916. doi: 10.1083/jcb.105.4.1901. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Couchman J. R., Rees D. A. The behaviour of fibroblasts migrating from chick heart explants: changes in adhesion, locomotion and growth, and in the distribution of actomyosin and fibronectin. J Cell Sci. 1979 Oct;39:149–165. doi: 10.1242/jcs.39.1.149. [DOI] [PubMed] [Google Scholar]
  10. Couchman J. R., Woods A., Hök M., Christner J. E. Characterization of a dermatan sulfate proteoglycan synthesized by murine parietal yolk sac (PYS-2) cells. J Biol Chem. 1985 Nov 5;260(25):13755–13762. [PubMed] [Google Scholar]
  11. Damsky C. H., Werb Z. Signal transduction by integrin receptors for extracellular matrix: cooperative processing of extracellular information. Curr Opin Cell Biol. 1992 Oct;4(5):772–781. doi: 10.1016/0955-0674(92)90100-q. [DOI] [PubMed] [Google Scholar]
  12. 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]
  13. 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]
  14. Fath K. R., Edgell C. J., Burridge K. The distribution of distinct integrins in focal contacts is determined by the substratum composition. J Cell Sci. 1989 Jan;92(Pt 1):67–75. doi: 10.1242/jcs.92.1.67. [DOI] [PubMed] [Google Scholar]
  15. Gould S. E., Upholt W. B., Kosher R. A. Syndecan 3: a member of the syndecan family of membrane-intercalated proteoglycans that is expressed in high amounts at the onset of chicken limb cartilage differentiation. Proc Natl Acad Sci U S A. 1992 Apr 15;89(8):3271–3275. doi: 10.1073/pnas.89.8.3271. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Hayashi Y., Haimovich B., Reszka A., Boettiger D., Horwitz A. Expression and function of chicken integrin beta 1 subunit and its cytoplasmic domain mutants in mouse NIH 3T3 cells. J Cell Biol. 1990 Jan;110(1):175–184. doi: 10.1083/jcb.110.1.175. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Hynes R. O. Integrins: versatility, modulation, and signaling in cell adhesion. Cell. 1992 Apr 3;69(1):11–25. doi: 10.1016/0092-8674(92)90115-s. [DOI] [PubMed] [Google Scholar]
  18. Kjellén L., Lindahl U. Proteoglycans: structures and interactions. Annu Rev Biochem. 1991;60:443–475. doi: 10.1146/annurev.bi.60.070191.002303. [DOI] [PubMed] [Google Scholar]
  19. 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]
  20. Kojima T., Shworak N. W., Rosenberg R. D. Molecular cloning and expression of two distinct cDNA-encoding heparan sulfate proteoglycan core proteins from a rat endothelial cell line. J Biol Chem. 1992 Mar 5;267(7):4870–4877. [PubMed] [Google Scholar]
  21. Kornberg L., Earp H. S., Parsons J. T., Schaller M., Juliano R. L. Cell adhesion or integrin clustering increases phosphorylation of a focal adhesion-associated tyrosine kinase. J Biol Chem. 1992 Nov 25;267(33):23439–23442. [PubMed] [Google Scholar]
  22. LeBaron R. G., Esko J. D., Woods A., Johansson S., Hök M. Adhesion of glycosaminoglycan-deficient chinese hamster ovary cell mutants to fibronectin substrata. J Cell Biol. 1988 Mar;106(3):945–952. doi: 10.1083/jcb.106.3.945. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. 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]
  24. 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]
  25. Marcantonio E. E., Guan J. L., Trevithick J. E., Hynes R. O. Mapping of the functional determinants of the integrin beta 1 cytoplasmic domain by site-directed mutagenesis. Cell Regul. 1990 Jul;1(8):597–604. doi: 10.1091/mbc.1.8.597. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Marynen P., Zhang J., Cassiman J. J., Van den Berghe H., David G. Partial primary structure of the 48- and 90-kilodalton core proteins of cell surface-associated heparan sulfate proteoglycans of lung fibroblasts. Prediction of an integral membrane domain and evidence for multiple distinct core proteins at the cell surface of human lung fibroblasts. J Biol Chem. 1989 Apr 25;264(12):7017–7024. [PubMed] [Google Scholar]
  27. Newman P. J., Allen R. W., Kahn R. A., Kunicki T. J. Quantitation of membrane glycoprotein IIIa on intact human platelets using the monoclonal antibody, AP-3. Blood. 1985 Jan;65(1):227–232. [PubMed] [Google Scholar]
  28. Pierce A., Lyon M., Hampson I. N., Cowling G. J., Gallagher J. T. Molecular cloning of the major cell surface heparan sulfate proteoglycan from rat liver. J Biol Chem. 1992 Feb 25;267(6):3894–3900. [PubMed] [Google Scholar]
  29. 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]
  30. Reszka A. A., Hayashi Y., Horwitz A. F. Identification of amino acid sequences in the integrin beta 1 cytoplasmic domain implicated in cytoskeletal association. J Cell Biol. 1992 Jun;117(6):1321–1330. doi: 10.1083/jcb.117.6.1321. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Saunders S., Jalkanen M., O'Farrell S., Bernfield M. Molecular cloning of syndecan, an integral membrane proteoglycan. J Cell Biol. 1989 Apr;108(4):1547–1556. doi: 10.1083/jcb.108.4.1547. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Tanaka Y., Adams D. H., Shaw S. Proteoglycans on endothelial cells present adhesion-inducing cytokines to leukocytes. Immunol Today. 1993 Mar;14(3):111–115. doi: 10.1016/0167-5699(93)90209-4. [DOI] [PubMed] [Google Scholar]
  33. Woods A., Couchman J. R. Focal adhesions and cell-matrix interactions. Coll Relat Res. 1988 Mar;8(2):155–182. doi: 10.1016/s0174-173x(88)80027-x. [DOI] [PubMed] [Google Scholar]
  34. Woods A., Couchman J. R. Heparan sulfate proteoglycans and signalling in cell adhesion. Adv Exp Med Biol. 1992;313:87–96. doi: 10.1007/978-1-4899-2444-5_9. [DOI] [PubMed] [Google Scholar]
  35. Woods A., Couchman J. R., Hök M. Heparan sulfate proteoglycans of rat embryo fibroblasts. A hydrophobic form may link cytoskeleton and matrix components. J Biol Chem. 1985 Sep 5;260(19):10872–10879. [PubMed] [Google Scholar]
  36. Woods A., Couchman J. R., Johansson S., Hök M. Adhesion and cytoskeletal organisation of fibroblasts in response to fibronectin fragments. EMBO J. 1986 Apr;5(4):665–670. doi: 10.1002/j.1460-2075.1986.tb04265.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  37. Woods A., Couchman J. R. Protein kinase C involvement in focal adhesion formation. J Cell Sci. 1992 Feb;101(Pt 2):277–290. doi: 10.1242/jcs.101.2.277. [DOI] [PubMed] [Google Scholar]
  38. Woods A., Hök M., Kjellén L., Smith C. G., Rees D. A. Relationship of heparan sulfate proteoglycans to the cytoskeleton and extracellular matrix of cultured fibroblasts. J Cell Biol. 1984 Nov;99(5):1743–1753. doi: 10.1083/jcb.99.5.1743. [DOI] [PMC free article] [PubMed] [Google Scholar]
  39. Woods A., McCarthy J. B., Furcht L. T., Couchman J. R. A synthetic peptide from the COOH-terminal heparin-binding domain of fibronectin promotes focal adhesion formation. Mol Biol Cell. 1993 Jun;4(6):605–613. doi: 10.1091/mbc.4.6.605. [DOI] [PMC free article] [PubMed] [Google Scholar]
  40. Yatohgo T., Izumi M., Kashiwagi H., Hayashi M. Novel purification of vitronectin from human plasma by heparin affinity chromatography. Cell Struct Funct. 1988 Aug;13(4):281–292. doi: 10.1247/csf.13.281. [DOI] [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]

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