Skip to main content
The Journal of Cell Biology logoLink to The Journal of Cell Biology
. 1983 Jun 1;96(6):1820–1823. doi: 10.1083/jcb.96.6.1820

Correlation between cell substrate attachment in vitro and cell surface heparan sulfate affinity for fibronectin and collagen

PMCID: PMC2112460  PMID: 6222058

Abstract

Heparan sulfate glycosaminoglycan, isolated from the cell surface of nonadhering murine myeloma cells (P3X63-Ag8653), does not bind to plasma fibronectin, but binds partially to collagen type I, as assayed by affinity chromatography with proteins immobilized on cyanogen bromide-activated Sepharose 4B. Identical results were obtained when myeloma heparan sulfate was cochromatographed, on the same fibronectin and collagen columns, with cell surface heparan sulfates collagen columns, with cell surface heparan sulfates from adhering Swiss mouse 3T3 and SV3T3 cells. These latter heparan sulfates do, however, bind to both fibronectin and collagen, as reported earlier (Stamatoglou, S.C., and J.M. Keller, 1981, Biochim. Biophys. Acta., 719:90-97). Cell adhesion assays established that hydrated collagen substrata can support myeloma cell attachment, but fibronectin cannot. Saturation of the heparan sulfate binding sites on the collagen substrata with heparan sulfate or heparin, prior to cell inoculation, abolished the ability to support cell adhesion, whereas chondroitin 4 sulfate, chondroitin 6 sulfate, and hyaluronic acid had no effect.

Full Text

The Full Text of this article is available as a PDF (487.9 KB).

Selected References

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

  1. BITTER T., MUIR H. M. A modified uronic acid carbazole reaction. Anal Biochem. 1962 Oct;4:330–334. doi: 10.1016/0003-2697(62)90095-7. [DOI] [PubMed] [Google Scholar]
  2. Culp L. A., Rollins B. J., Buniel J., Hitri S. Two functionally distinct pools of glycosaminoglycan in the substrate adhesion site of murine cells. J Cell Biol. 1978 Dec;79(3):788–801. doi: 10.1083/jcb.79.3.788. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Culp L. A. Substrate-attached glycoproteins mediating adhesion of normal and virus-transformed mouse fibroblasts. J Cell Biol. 1974 Oct;63(1):71–83. doi: 10.1083/jcb.63.1.71. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Fransson L. A., Sjöberg I., Havsmark B. Structural studies on heparan sulphates. Characterization of oligosaccharides; obtained by periodate oxidation and alkaline elimination. Eur J Biochem. 1980 May;106(1):59–69. [PubMed] [Google Scholar]
  5. Hedman K., Johansson S., Vartio T., Kjellén L., Vaheri A., Hök M. Structure of the pericellular matrix: association of heparan and chondroitin sulfates with fibronectin-procollagen fibers. Cell. 1982 Mar;28(3):663–671. doi: 10.1016/0092-8674(82)90221-5. [DOI] [PubMed] [Google Scholar]
  6. Hughes R. C., Butters T. D., Aplin J. D. Cell surface molecules involved in fibronectin-mediated adhesion. A study using specific antisera. Eur J Cell Biol. 1981 Dec;26(1):198–207. [PubMed] [Google Scholar]
  7. Johnston L. S., Keller K. L., Keller J. M. The heparan sulfates of Swiss mouse 3T3 cells. The effect of transformation. Biochim Biophys Acta. 1979 Feb 19;583(1):81–94. doi: 10.1016/0304-4165(79)90312-x. [DOI] [PubMed] [Google Scholar]
  8. Keller K. L., Keller J. M., Moy J. N. Heparan sulfates from Swiss mouse 3T3 and SV3T3 cells: O-sulfate difference. Biochemistry. 1980 May 27;19(11):2529–2536. doi: 10.1021/bi00552a035. [DOI] [PubMed] [Google Scholar]
  9. Kleinman H. K., Hewitt A. T., Murray J. C., Liotta L. A., Rennard S. I., Pennypacker J. P., McGoodwin E. B., Martin G. R., Fishman P. H. Cellular and metabolic specificity in the interaction of adhesion proteins with collagen and with cells. J Supramol Struct. 1979;11(1):69–78. doi: 10.1002/jss.400110108. [DOI] [PubMed] [Google Scholar]
  10. Kraemer P. M. Heparan sulfates of cultured cells. II. Acid-soluble and -precipitable species of different cell lines. Biochemistry. 1971 Apr 13;10(8):1445–1451. doi: 10.1021/bi00784a027. [DOI] [PubMed] [Google Scholar]
  11. Laemmli U. K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. doi: 10.1038/227680a0. [DOI] [PubMed] [Google Scholar]
  12. Laterra J., Ansbacher R., Culp L. A. Glycosaminoglycans that bind cold-insoluble globulin in cell-substratum adhesion sites of murine fibroblasts. Proc Natl Acad Sci U S A. 1980 Nov;77(11):6662–6666. doi: 10.1073/pnas.77.11.6662. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Laterra J., Silbert J. E., Culp L. A. Cell surface heparan sulfate mediates some adhesive responses to glycosaminoglycan-binding matrices, including fibronectin. J Cell Biol. 1983 Jan;96(1):112–123. doi: 10.1083/jcb.96.1.112. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Lindahl U., Hök M. Glycosaminoglycans and their binding to biological macromolecules. Annu Rev Biochem. 1978;47:385–417. doi: 10.1146/annurev.bi.47.070178.002125. [DOI] [PubMed] [Google Scholar]
  15. Norling B., Glimelius B., Wasteson A. Heparan sulfate proteoglycan of cultured cells: demonstration of a lipid- and a matrix-associated form. Biochem Biophys Res Commun. 1981 Dec 31;103(4):1265–1272. doi: 10.1016/0006-291x(81)90259-x. [DOI] [PubMed] [Google Scholar]
  16. Pierschbacher M. D., Hayman E. G., Ruoslahti E. Location of the cell-attachment site in fibronectin with monoclonal antibodies and proteolytic fragments of the molecule. Cell. 1981 Oct;26(2 Pt 2):259–267. doi: 10.1016/0092-8674(81)90308-1. [DOI] [PubMed] [Google Scholar]
  17. Rollins B. J., Culp L. A. Glycosaminoglycans in the substrate adhesion sites of normal and virus-transformed murine cells. Biochemistry. 1979 Jan 9;18(1):141–148. doi: 10.1021/bi00568a022. [DOI] [PubMed] [Google Scholar]
  18. Rubin K., Oldberg A., Hök M., Obrink B. Adhesion of rat hepatocytes to collagen. Exp Cell Res. 1978 Nov;117(1):165–177. doi: 10.1016/0014-4827(78)90439-1. [DOI] [PubMed] [Google Scholar]
  19. Ruoslahti E., Hayman E. G., Pierschbacher M., Engvall E. Fibronectin: purification, immunochemical properties, and biological activities. Methods Enzymol. 1982;82(Pt A):803–831. doi: 10.1016/0076-6879(82)82103-4. [DOI] [PubMed] [Google Scholar]
  20. Schubert D., LaCorbiere M. A role of secreted glycosaminoglycans in cell-substratum adhesion. J Biol Chem. 1980 Dec 10;255(23):11564–11569. [PubMed] [Google Scholar]
  21. Stamatoglou S. C., Keller J. M. Interactions of cellular glycosaminoglycans with plasma fibronectin and collagen. Biochim Biophys Acta. 1982 Oct 28;719(1):90–97. doi: 10.1016/0304-4165(82)90311-7. [DOI] [PubMed] [Google Scholar]
  22. Underhill C. B., Keller J. M. A transformation-dependent difference in the heparan sulfate associated with the cell surface. Biochem Biophys Res Commun. 1975 Mar 17;63(2):448–454. doi: 10.1016/0006-291x(75)90708-1. [DOI] [PubMed] [Google Scholar]
  23. Winterbourne D. J., Mora P. T. Cells selected for high tumorigenicity or transformed by simian virus 40 synthesize heparan sulfate with reduced degree of sulfation. J Biol Chem. 1981 May 10;256(9):4310–4320. [PubMed] [Google Scholar]

Articles from The Journal of Cell Biology are provided here courtesy of The Rockefeller University Press

RESOURCES