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. 1986 Feb 1;102(2):403–412. doi: 10.1083/jcb.102.2.403

A heparin-binding domain from N-CAM is involved in neural cell- substratum adhesion

PMCID: PMC2114085  PMID: 2418031

Abstract

Cell-substratum adhesion in the embryonic chicken nervous system has been shown to be mediated in part by a 170,000-mol-wt polypeptide that is a component of adherons. Attachment of retinal cells to the 170,000- mol-wt protein is inhibited by the C1H3 monoclonal antibody and by heparan sulfate (Cole, G. J., D. Schubert, and L. Glaser, 1985, J. Cell Biol., 100:1192-1199). In the present study we have demonstrated that the 170,000-mol-wt C1H3 polypeptide is immunologically identical to the neural cell adhesion molecule N-CAM, and that the 170,000-mol-wt component of N-CAM is preferentially secreted by cells as a component of adherons. We have identified a monoclonal antibody, designated B1A3, that inhibits heparin binding to N-CAM and cell-to-substratum adhesion. A 25,000-mol-wt heparin (heparan sulfate)-binding domain of N-CAM has been identified by limited proteolysis, and this fragment promotes cell attachment when bound to glass surfaces. The fragment also partially inhibits cell binding to adherons when bound to retinal cells, and the B1A3 monoclonal antibody inhibits retinal cell attachment to substrata composed of intact N-CAM or the heparin-binding domain. These data are the first evidence that N-CAM is a multifunctional protein that contains both cell-and heparin (heparan sulfate)-binding domains.

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Selected References

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  1. Bottenstein J. E., Sato G. H. Growth of a rat neuroblastoma cell line in serum-free supplemented medium. Proc Natl Acad Sci U S A. 1979 Jan;76(1):514–517. doi: 10.1073/pnas.76.1.514. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Boucaut J. C., Darribère T., Poole T. J., Aoyama H., Yamada K. M., Thiery J. P. Biologically active synthetic peptides as probes of embryonic development: a competitive peptide inhibitor of fibronectin function inhibits gastrulation in amphibian embryos and neural crest cell migration in avian embryos. J Cell Biol. 1984 Nov;99(5):1822–1830. doi: 10.1083/jcb.99.5.1822. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Cole G. J., Glaser L. Cell-substratum adhesion in embryonic chick central nervous system is mediated by a 170,000-mol-wt neural-specific polypeptide. J Cell Biol. 1984 Nov;99(5):1605–1612. doi: 10.1083/jcb.99.5.1605. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Cole G. J., Glaser L. Identification of novel neural- and neural retina-specific antigens with a monoclonal antibody. Proc Natl Acad Sci U S A. 1984 Apr;81(7):2260–2264. doi: 10.1073/pnas.81.7.2260. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Cole G. J., Glaser L. Inhibition of embryonic neural retina cell-substratum adhesion with a monoclonal antibody. J Biol Chem. 1984 Apr 10;259(7):4031–4034. [PubMed] [Google Scholar]
  6. Cole G. J., Schubert D., Glaser L. Cell-substratum adhesion in chick neural retina depends upon protein-heparan sulfate interactions. J Cell Biol. 1985 Apr;100(4):1192–1199. doi: 10.1083/jcb.100.4.1192. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Edelman G. M. Cell adhesion molecules. Science. 1983 Feb 4;219(4584):450–457. doi: 10.1126/science.6823544. [DOI] [PubMed] [Google Scholar]
  8. Edgar D., Timpl R., Thoenen H. The heparin-binding domain of laminin is responsible for its effects on neurite outgrowth and neuronal survival. EMBO J. 1984 Jul;3(7):1463–1468. doi: 10.1002/j.1460-2075.1984.tb01997.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Friedlander D. R., Brackenbury R., Edelman G. M. Conversion of embryonic form to adult forms of N-CAM in vitro: results from de novo synthesis of adult forms. J Cell Biol. 1985 Aug;101(2):412–419. doi: 10.1083/jcb.101.2.412. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Grumet M., Hoffman S., Edelman G. M. Two antigenically related neuronal cell adhesion molecules of different specificities mediate neuron-neuron and neuron-glia adhesion. Proc Natl Acad Sci U S A. 1984 Jan;81(1):267–271. doi: 10.1073/pnas.81.1.267. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Grumet M., Rutishauser U., Edelman G. M. Neural cell adhesion molecule is on embryonic muscle cells and mediates adhesion to nerve cells in vitro. Nature. 1982 Feb 25;295(5851):693–695. doi: 10.1038/295693a0. [DOI] [PubMed] [Google Scholar]
  12. Grunwald G. B., Pratt R. S., Lilien J. Enzymic dissection of embryonic cell adhesive mechanisms. III. Immunological identification of a component of the calcium-dependent adhesive system of embryonic chick neural retina cells. J Cell Sci. 1982 Jun;55:69–83. doi: 10.1242/jcs.55.1.69. [DOI] [PubMed] [Google Scholar]
  13. Hausman R. E., Moscona A. A. Isolation of retina-specific cell-aggregating factor from membranes of embryonic neural retina tissue. Proc Natl Acad Sci U S A. 1976 Oct;73(10):3594–3598. doi: 10.1073/pnas.73.10.3594. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Hirn M., Ghandour M. S., Deagostini-Bazin H., Goridis C. Molecular heterogeneity and structural evolution during cerebellar ontogeny detected by monoclonal antibody of the mouse cell surface antigen BSP-2. Brain Res. 1983 Apr 11;265(1):87–100. doi: 10.1016/0006-8993(83)91337-9. [DOI] [PubMed] [Google Scholar]
  15. Hynes R. O., Yamada K. M. Fibronectins: multifunctional modular glycoproteins. J Cell Biol. 1982 Nov;95(2 Pt 1):369–377. doi: 10.1083/jcb.95.2.369. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Johansson S., Hök M. Substrate adhesion of rat hepatocytes: on the mechanism of attachment to fibronectin. J Cell Biol. 1984 Mar;98(3):810–817. doi: 10.1083/jcb.98.3.810. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Jørgensen O. S., Delouvée A., Thiery J. P., Edelman G. M. The nervous system specific protein D2 is involved in adhesion among neurites from cultured rat ganglia. FEBS Lett. 1980 Feb 25;111(1):39–42. doi: 10.1016/0014-5793(80)80756-3. [DOI] [PubMed] [Google Scholar]
  18. Keilhauer G., Faissner A., Schachner M. Differential inhibition of neurone-neurone, neurone-astrocyte and astrocyte-astrocyte adhesion by L1, L2 and N-CAM antibodies. Nature. 1985 Aug 22;316(6030):728–730. doi: 10.1038/316728a0. [DOI] [PubMed] [Google Scholar]
  19. Lander A. D., Fujii D. K., Reichardt L. F. Laminin is associated with the "neurite outgrowth-promoting factors" found in conditioned media. Proc Natl Acad Sci U S A. 1985 Apr;82(7):2183–2187. doi: 10.1073/pnas.82.7.2183. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Noble M., Albrechtsen M., Møller C., Lyles J., Bock E., Goridis C., Watanabe M., Rutishauser U. Glial cells express N-CAM/D2-CAM-like polypeptides in vitro. Nature. 1985 Aug 22;316(6030):725–728. doi: 10.1038/316725a0. [DOI] [PubMed] [Google Scholar]
  21. Osterlund E., Eronen I., Osterlund K., Vuento M. Secondary structure of human plasma fibronectin: conformational change induced by calf alveolar heparan sulfates. Biochemistry. 1985 May 21;24(11):2661–2667. doi: 10.1021/bi00332a011. [DOI] [PubMed] [Google Scholar]
  22. 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]
  23. Rathjen F. G., Schachner M. Immunocytological and biochemical characterization of a new neuronal cell surface component (L1 antigen) which is involved in cell adhesion. EMBO J. 1984 Jan;3(1):1–10. doi: 10.1002/j.1460-2075.1984.tb01753.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Rogers S. L., McCarthy J. B., Palm S. L., Furcht L. T., Letourneau P. C. Neuron-specific interactions with two neurite-promoting fragments of fibronectin. J Neurosci. 1985 Feb;5(2):369–378. doi: 10.1523/JNEUROSCI.05-02-00369.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Ruoslahti E., Hayman E. G., Engvall E., Cothran W. C., Butler W. T. Alignment of biologically active domains in the fibronectin molecule. J Biol Chem. 1981 Jul 25;256(14):7277–7281. [PubMed] [Google Scholar]
  26. Rutishauser U. Developmental biology of a neural cell adhesion molecule. Nature. 1984 Aug 16;310(5978):549–554. doi: 10.1038/310549a0. [DOI] [PubMed] [Google Scholar]
  27. Rutishauser U., Hoffman S., Edelman G. M. Binding properties of a cell adhesion molecule from neural tissue. Proc Natl Acad Sci U S A. 1982 Jan;79(2):685–689. doi: 10.1073/pnas.79.2.685. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Schubert D., LaCorbiere M. Isolation of a cell-surface receptor for chick neural retina adherons. J Cell Biol. 1985 Jan;100(1):56–63. doi: 10.1083/jcb.100.1.56. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Schubert D., LaCorbiere M. Isolation of an adhesion-mediating protein from chick neural retina adherons. J Cell Biol. 1985 Sep;101(3):1071–1077. doi: 10.1083/jcb.101.3.1071. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Schubert D., LaCorbiere M., Klier F. G., Birdwell C. A role for adherons in neural retina cell adhesion. J Cell Biol. 1983 Apr;96(4):990–998. doi: 10.1083/jcb.96.4.990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Silver J., Rutishauser U. Guidance of optic axons in vivo by a preformed adhesive pathway on neuroepithelial endfeet. Dev Biol. 1984 Dec;106(2):485–499. doi: 10.1016/0012-1606(84)90248-3. [DOI] [PubMed] [Google Scholar]
  32. Thiery J. P., Brackenbury R., Rutishauser U., Edelman G. M. Adhesion among neural cells of the chick embryo. II. Purification and characterization of a cell adhesion molecule from neural retina. J Biol Chem. 1977 Oct 10;252(19):6841–6845. [PubMed] [Google Scholar]
  33. Unsicker K., Skaper S. D., Davis G. E., Manthorpe M., Varon S. Comparison of the effects of laminin and the polyornithine-binding neurite promoting factor from RN22 Schwannoma cells on neurite regeneration from cultured newborn and adult rat dorsal root ganglion neurons. Brain Res. 1985 Jan;349(1-2):304–308. doi: 10.1016/0165-3806(85)90161-0. [DOI] [PubMed] [Google Scholar]
  34. Williams R. K., Goridis C., Akeson R. Individual neural cell types express immunologically distinct N-CAM forms. J Cell Biol. 1985 Jul;101(1):36–42. doi: 10.1083/jcb.101.1.36. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. Witte D., Gottlieb D. I. Time of appearance and tissue distribution of a cell surface antigen in early chick development. Brain Res. 1983 Jul;285(1):63–67. doi: 10.1016/0165-3806(83)90109-8. [DOI] [PubMed] [Google Scholar]
  36. Yamada K. M. Cell surface interactions with extracellular materials. Annu Rev Biochem. 1983;52:761–799. doi: 10.1146/annurev.bi.52.070183.003553. [DOI] [PubMed] [Google Scholar]
  37. Yamada K. M., Kennedy D. W., Kimata K., Pratt R. M. Characterization of fibronectin interactions with glycosaminoglycans and identification of active proteolytic fragments. J Biol Chem. 1980 Jul 10;255(13):6055–6063. [PubMed] [Google Scholar]

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