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. 1992 Oct 1;119(1):191–202. doi: 10.1083/jcb.119.1.191

Homophilic and heterophilic binding activities of Nr-CAM, a nervous system cell adhesion molecule

PMCID: PMC2289630  PMID: 1527169

Abstract

Nr-CAM is a membrane glycoprotein that is expressed on neurons. It is structurally related to members of the N-CAM superfamily of neural cell adhesion molecules having six immunoglobulin-like domains and five fibronectin type III repeats in the extracellular region. We have found that the aggregation of chick brain cells was inhibited by anti-Nr-CAM Fab' fragments, indicating that Nr-CAM can act as a cell adhesion molecule. To clarify the mode of action of Nr-CAM, a mouse fibroblast cell line L-M(TK-) (or L cells) was transfected with a DNA expression construct encoding an entire chicken Nr-CAM cDNA sequence. After transfection, L cells expressed Nr-CAM on their surface and aggregated. Aggregation was specifically inhibited by anti-Nr-CAM Fab' fragments. To check the specificity of this aggregation, a fusion protein (FGTNr) consisting of glutathione S-transferase linked to the six immunoglobulin domains and the first fibronectin type III repeat of Nr- CAM was expressed in Escherichia coli. Addition of FGTNr to the transfected cells blocked their aggregation. Further analysis using a combination of cell aggregation assays, binding of cells to FGTNr- coated substrates, aggregation of FGTNr-coated Covaspheres and binding of FGTNr-coated Covaspheres to FGTNr-coated substrates revealed that Nr- CAM mediates two types of cell interactions: a homophilic, divalent cation-independent binding, and a heterophilic, divalent cation- dependent binding. Homophilic binding was demonstrated between transfected L cells, between chick embryo brain cells and FGTNr, and between Covaspheres to which FGTNr was covalently attached. Heterophilic binding was shown to occur between transfected and untransfected L cells, and between FGTNr and primary chick embryo fibroblasts; in all cases, it was dependent on the presence of either calcium or magnesium. Primary chick embryo glia or a human glial cell line did not bind to FGTNr-coated substrates. The results indicate that Nr-CAM is a cell adhesion molecule of the nervous system that can bind by two distinct mechanisms, a homophilic mechanism that can mediate interactions between neurons and a heterophilic mechanism that can mediate binding between neurons and other cells such as fibroblasts.

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Selected References

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  1. Afar D. E., Marius R. M., Salzer J. L., Stanners C. P., Braun P. E., Bell J. C. Cell adhesion properties of myelin-associated glycoprotein in L cell fibroblasts. J Neurosci Res. 1991 Aug;29(4):429–436. doi: 10.1002/jnr.490290403. [DOI] [PubMed] [Google Scholar]
  2. Albelda S. M., Buck C. A. Integrins and other cell adhesion molecules. FASEB J. 1990 Aug;4(11):2868–2880. [PubMed] [Google Scholar]
  3. Albelda S. M., Muller W. A., Buck C. A., Newman P. J. Molecular and cellular properties of PECAM-1 (endoCAM/CD31): a novel vascular cell-cell adhesion molecule. J Cell Biol. 1991 Sep;114(5):1059–1068. doi: 10.1083/jcb.114.5.1059. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Arquint M., Roder J., Chia L. S., Down J., Wilkinson D., Bayley H., Braun P., Dunn R. Molecular cloning and primary structure of myelin-associated glycoprotein. Proc Natl Acad Sci U S A. 1987 Jan;84(2):600–604. doi: 10.1073/pnas.84.2.600. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Bieber A. J., Snow P. M., Hortsch M., Patel N. H., Jacobs J. R., Traquina Z. R., Schilling J., Goodman C. S. Drosophila neuroglian: a member of the immunoglobulin superfamily with extensive homology to the vertebrate neural adhesion molecule L1. Cell. 1989 Nov 3;59(3):447–460. doi: 10.1016/0092-8674(89)90029-9. [DOI] [PubMed] [Google Scholar]
  6. Brackenbury R., Rutishauser U., Edelman G. M. Distinct calcium-independent and calcium-dependent adhesion systems of chicken embryo cells. Proc Natl Acad Sci U S A. 1981 Jan;78(1):387–391. doi: 10.1073/pnas.78.1.387. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Brackenbury R., Thiery J. P., Rutishauser U., Edelman G. M. Adhesion among neural cells of the chick embryo. I. An immunological assay for molecules involved in cell-cell binding. J Biol Chem. 1977 Oct 10;252(19):6835–6840. [PubMed] [Google Scholar]
  8. Brümmendorf T., Wolff J. M., Frank R., Rathjen F. G. Neural cell recognition molecule F11: homology with fibronectin type III and immunoglobulin type C domains. Neuron. 1989 Apr;2(4):1351–1361. doi: 10.1016/0896-6273(89)90073-1. [DOI] [PubMed] [Google Scholar]
  9. Burgoon M. P., Grumet M., Mauro V., Edelman G. M., Cunningham B. A. Structure of the chicken neuron-glia cell adhesion molecule, Ng-CAM: origin of the polypeptides and relation to the Ig superfamily. J Cell Biol. 1991 Mar;112(5):1017–1029. doi: 10.1083/jcb.112.5.1017. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Crossin K. L., Chuong C. M., Edelman G. M. Expression sequences of cell adhesion molecules. Proc Natl Acad Sci U S A. 1985 Oct;82(20):6942–6946. doi: 10.1073/pnas.82.20.6942. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. D'Urso D., Brophy P. J., Staugaitis S. M., Gillespie C. S., Frey A. B., Stempak J. G., Colman D. R. Protein zero of peripheral nerve myelin: biosynthesis, membrane insertion, and evidence for homotypic interaction. Neuron. 1990 Mar;4(3):449–460. doi: 10.1016/0896-6273(90)90057-m. [DOI] [PubMed] [Google Scholar]
  12. Daniloff J. K., Chuong C. M., Levi G., Edelman G. M. Differential distribution of cell adhesion molecules during histogenesis of the chick nervous system. J Neurosci. 1986 Mar;6(3):739–758. doi: 10.1523/JNEUROSCI.06-03-00739.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Dulac C., Tropak M. B., Cameron-Curry P., Rossier J., Marshak D. R., Roder J., Le Douarin N. M. Molecular characterization of the Schwann cell myelin protein, SMP: structural similarities within the immunoglobulin superfamily. Neuron. 1992 Feb;8(2):323–334. doi: 10.1016/0896-6273(92)90298-r. [DOI] [PubMed] [Google Scholar]
  14. Edelman G. M., Crossin K. L. Cell adhesion molecules: implications for a molecular histology. Annu Rev Biochem. 1991;60:155–190. doi: 10.1146/annurev.bi.60.070191.001103. [DOI] [PubMed] [Google Scholar]
  15. Edelman G. M., Murray B. A., Mege R. M., Cunningham B. A., Gallin W. J. Cellular expression of liver and neural cell adhesion molecules after transfection with their cDNAs results in specific cell-cell binding. Proc Natl Acad Sci U S A. 1987 Dec;84(23):8502–8506. doi: 10.1073/pnas.84.23.8502. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Edelman G. M. Surface modulation in cell recognition and cell growth. Science. 1976 Apr 16;192(4236):218–226. doi: 10.1126/science.769162. [DOI] [PubMed] [Google Scholar]
  17. Elices M. J., Osborn L., Takada Y., Crouse C., Luhowskyj S., Hemler M. E., Lobb R. R. VCAM-1 on activated endothelium interacts with the leukocyte integrin VLA-4 at a site distinct from the VLA-4/fibronectin binding site. Cell. 1990 Feb 23;60(4):577–584. doi: 10.1016/0092-8674(90)90661-w. [DOI] [PubMed] [Google Scholar]
  18. Filbin M. T., Walsh F. S., Trapp B. D., Pizzey J. A., Tennekoon G. I. Role of myelin P0 protein as a homophilic adhesion molecule. Nature. 1990 Apr 26;344(6269):871–872. doi: 10.1038/344871a0. [DOI] [PubMed] [Google Scholar]
  19. Fischer G., Künemund V., Schachner M. Neurite outgrowth patterns in cerebellar microexplant cultures are affected by antibodies to the cell surface glycoprotein L1. J Neurosci. 1986 Feb;6(2):605–612. doi: 10.1523/JNEUROSCI.06-02-00605.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Friedlander D. R., Hoffman S., Edelman G. M. Functional mapping of cytotactin: proteolytic fragments active in cell-substrate adhesion. J Cell Biol. 1988 Dec;107(6 Pt 1):2329–2340. doi: 10.1083/jcb.107.6.2329. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Furley A. J., Morton S. B., Manalo D., Karagogeos D., Dodd J., Jessell T. M. The axonal glycoprotein TAG-1 is an immunoglobulin superfamily member with neurite outgrowth-promoting activity. Cell. 1990 Apr 6;61(1):157–170. doi: 10.1016/0092-8674(90)90223-2. [DOI] [PubMed] [Google Scholar]
  22. Gennarini G., Cibelli G., Rougon G., Mattei M. G., Goridis C. The mouse neuronal cell surface protein F3: a phosphatidylinositol-anchored member of the immunoglobulin superfamily related to chicken contactin. J Cell Biol. 1989 Aug;109(2):775–788. doi: 10.1083/jcb.109.2.775. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Gorman C. M., Howard B. H., Reeves R. Expression of recombinant plasmids in mammalian cells is enhanced by sodium butyrate. Nucleic Acids Res. 1983 Nov 11;11(21):7631–7648. doi: 10.1093/nar/11.21.7631. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Grumet M., Edelman G. M. Heterotypic binding between neuronal membrane vesicles and glial cells is mediated by a specific cell adhesion molecule. J Cell Biol. 1984 May;98(5):1746–1756. doi: 10.1083/jcb.98.5.1746. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Grumet M., Edelman G. M. Neuron-glia cell adhesion molecule interacts with neurons and astroglia via different binding mechanisms. J Cell Biol. 1988 Feb;106(2):487–503. doi: 10.1083/jcb.106.2.487. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Grumet M., Hoffman S., Chuong C. M., Edelman G. M. Polypeptide components and binding functions of neuron-glia cell adhesion molecules. Proc Natl Acad Sci U S A. 1984 Dec;81(24):7989–7993. doi: 10.1073/pnas.81.24.7989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Grumet M., Mauro V., Burgoon M. P., Edelman G. M., Cunningham B. A. Structure of a new nervous system glycoprotein, Nr-CAM, and its relationship to subgroups of neural cell adhesion molecules. J Cell Biol. 1991 Jun;113(6):1399–1412. doi: 10.1083/jcb.113.6.1399. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Grumet M. Structure, expression, and function of Ng-CAM, a member of the immunoglobulin superfamily involved in neuron-neuron and neuron-glia adhesion. J Neurosci Res. 1992 Jan;31(1):1–13. doi: 10.1002/jnr.490310102. [DOI] [PubMed] [Google Scholar]
  29. Harrelson A. L., Goodman C. S. Growth cone guidance in insects: fasciclin II is a member of the immunoglobulin superfamily. Science. 1988 Nov 4;242(4879):700–708. doi: 10.1126/science.3187519. [DOI] [PubMed] [Google Scholar]
  30. Hoffman S., Crossin K. L., Edelman G. M. Molecular forms, binding functions, and developmental expression patterns of cytotactin and cytotactin-binding proteoglycan, an interactive pair of extracellular matrix molecules. J Cell Biol. 1988 Feb;106(2):519–532. doi: 10.1083/jcb.106.2.519. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Hoffman S., Edelman G. M. Kinetics of homophilic binding by embryonic and adult forms of the neural cell adhesion molecule. Proc Natl Acad Sci U S A. 1983 Sep;80(18):5762–5766. doi: 10.1073/pnas.80.18.5762. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Hoffman S., Friedlander D. R., Chuong C. M., Grumet M., Edelman G. M. Differential contributions of Ng-CAM and N-CAM to cell adhesion in different neural regions. J Cell Biol. 1986 Jul;103(1):145–158. doi: 10.1083/jcb.103.1.145. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Hortsch M., Bieber A. J., Patel N. H., Goodman C. S. Differential splicing generates a nervous system-specific form of Drosophila neuroglian. Neuron. 1990 May;4(5):697–709. doi: 10.1016/0896-6273(90)90196-m. [DOI] [PubMed] [Google Scholar]
  34. 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]
  35. Kuhn T. B., Stoeckli E. T., Condrau M. A., Rathjen F. G., Sonderegger P. Neurite outgrowth on immobilized axonin-1 is mediated by a heterophilic interaction with L1(G4). J Cell Biol. 1991 Nov;115(4):1113–1126. doi: 10.1083/jcb.115.4.1113. [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. Lemke G., Axel R. Isolation and sequence of a cDNA encoding the major structural protein of peripheral myelin. Cell. 1985 Mar;40(3):501–508. doi: 10.1016/0092-8674(85)90198-9. [DOI] [PubMed] [Google Scholar]
  37. Levi G., Crossin K. L., Edelman G. M. Expression sequences and distribution of two primary cell adhesion molecules during embryonic development of Xenopus laevis. J Cell Biol. 1987 Nov;105(5):2359–2372. doi: 10.1083/jcb.105.5.2359. [DOI] [PMC free article] [PubMed] [Google Scholar]
  38. Marlin S. D., Springer T. A. Purified intercellular adhesion molecule-1 (ICAM-1) is a ligand for lymphocyte function-associated antigen 1 (LFA-1). Cell. 1987 Dec 4;51(5):813–819. doi: 10.1016/0092-8674(87)90104-8. [DOI] [PubMed] [Google Scholar]
  39. Moos M., Tacke R., Scherer H., Teplow D., Früh K., Schachner M. Neural adhesion molecule L1 as a member of the immunoglobulin superfamily with binding domains similar to fibronectin. Nature. 1988 Aug 25;334(6184):701–703. doi: 10.1038/334701a0. [DOI] [PubMed] [Google Scholar]
  40. Muller W. A., Berman M. E., Newman P. J., DeLisser H. M., Albelda S. M. A heterophilic adhesion mechanism for platelet/endothelial cell adhesion molecule 1 (CD31). J Exp Med. 1992 May 1;175(5):1401–1404. doi: 10.1084/jem.175.5.1401. [DOI] [PMC free article] [PubMed] [Google Scholar]
  41. Oikawa S., Inuzuka C., Kuroki M., Arakawa F., Matsuoka Y., Kosaki G., Nakazato H. A specific heterotypic cell adhesion activity between members of carcinoembryonic antigen family, W272 and NCA, is mediated by N-domains. J Biol Chem. 1991 May 5;266(13):7995–8001. [PubMed] [Google Scholar]
  42. Prince J. T., Milona N., Stallcup W. B. Characterization of a partial cDNA clone for the NILE glycoprotein and identification of the encoded polypeptide domain. J Neurosci. 1989 Mar;9(3):876–883. doi: 10.1523/JNEUROSCI.09-03-00876.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  43. Ranscht B. Sequence of contactin, a 130-kD glycoprotein concentrated in areas of interneuronal contact, defines a new member of the immunoglobulin supergene family in the nervous system. J Cell Biol. 1988 Oct;107(4):1561–1573. doi: 10.1083/jcb.107.4.1561. [DOI] [PMC free article] [PubMed] [Google Scholar]
  44. Salzer J. L., Holmes W. P., Colman D. R. The amino acid sequences of the myelin-associated glycoproteins: homology to the immunoglobulin gene superfamily. J Cell Biol. 1987 Apr;104(4):957–965. doi: 10.1083/jcb.104.4.957. [DOI] [PMC free article] [PubMed] [Google Scholar]
  45. Schneider-Schaulies J., von Brunn A., Schachner M. Recombinant peripheral myelin protein P0 confers both adhesion and neurite outgrowth-promoting properties. J Neurosci Res. 1990 Nov;27(3):286–297. doi: 10.1002/jnr.490270307. [DOI] [PubMed] [Google Scholar]
  46. Smith D. B., Johnson K. S. Single-step purification of polypeptides expressed in Escherichia coli as fusions with glutathione S-transferase. Gene. 1988 Jul 15;67(1):31–40. doi: 10.1016/0378-1119(88)90005-4. [DOI] [PubMed] [Google Scholar]
  47. Staunton D. E., Marlin S. D., Stratowa C., Dustin M. L., Springer T. A. Primary structure of ICAM-1 demonstrates interaction between members of the immunoglobulin and integrin supergene families. Cell. 1988 Mar 25;52(6):925–933. doi: 10.1016/0092-8674(88)90434-5. [DOI] [PubMed] [Google Scholar]
  48. Tanaka H., Matsui T., Agata A., Tomura M., Kubota I., McFarland K. C., Kohr B., Lee A., Phillips H. S., Shelton D. L. Molecular cloning and expression of a novel adhesion molecule, SC1. Neuron. 1991 Oct;7(4):535–545. doi: 10.1016/0896-6273(91)90366-8. [DOI] [PubMed] [Google Scholar]
  49. Thiery J. P., Delouvée A., Grumet M., Edelman G. M. Initial appearance and regional distribution of the neuron-glia cell adhesion molecule in the chick embryo. J Cell Biol. 1985 Feb;100(2):442–456. doi: 10.1083/jcb.100.2.442. [DOI] [PMC free article] [PubMed] [Google Scholar]
  50. Thiery J. P., Duband J. L., Rutishauser U., Edelman G. M. Cell adhesion molecules in early chicken embryogenesis. Proc Natl Acad Sci U S A. 1982 Nov;79(21):6737–6741. doi: 10.1073/pnas.79.21.6737. [DOI] [PMC free article] [PubMed] [Google Scholar]
  51. Zuellig R. A., Rader C., Schroeder A., Kalousek M. B., Von Bohlen und Halbach F., Osterwalder T., Inan C., Stoeckli E. T., Affolter H. U., Fritz A. The axonally secreted cell adhesion molecule, axonin-1. Primary structure, immunoglobulin-like and fibronectin-type-III-like domains and glycosyl-phosphatidylinositol anchorage. Eur J Biochem. 1992 Mar 1;204(2):453–463. doi: 10.1111/j.1432-1033.1992.tb16655.x. [DOI] [PubMed] [Google Scholar]
  52. de la Rosa E. J., Kayyem J. F., Roman J. M., Stierhof Y. D., Dreyer W. J., Schwarz U. Topologically restricted appearance in the developing chick retinotectal system of Bravo, a neural surface protein: experimental modulation by environmental cues. J Cell Biol. 1990 Dec;111(6 Pt 2):3087–3096. doi: 10.1083/jcb.111.6.3087. [DOI] [PMC free article] [PubMed] [Google Scholar]

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