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. 1986 Nov 1;103(5):1729–1737. doi: 10.1083/jcb.103.5.1729

Topography of N-CAM structural and functional determinants. II. Placement of monoclonal antibody epitopes

PMCID: PMC2114365  PMID: 2430977

Abstract

The accompanying report (Watanabe, M., A. L. Frelinger III, and U. Rutishauser, 1986, J. Cell Biol., 103:1721-1727) describes a set of monoclonal antibodies (mAbs) directed against N-CAM epitopes representing the known major structural and functional domains of the molecule. In this study, we have generated and separated a variety of peptide fragments from N-CAM, and then used their size and reactivity with each antibody to position the antigenic sites along the peptide chain. This epitope map, together with the biological properties of the antibodies and previous studies on N-CAM, have been used to construct a topographical model for the molecule in the cell membrane.

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Selected References

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  1. Cole G. J., Glaser L. A heparin-binding domain from N-CAM is involved in neural cell-substratum adhesion. J Cell Biol. 1986 Feb;102(2):403–412. doi: 10.1083/jcb.102.2.403. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Crossin K. L., Edelman G. M., Cunningham B. A. Mapping of three carbohydrate attachment sites in embryonic and adult forms of the neural cell adhesion molecule. J Cell Biol. 1984 Nov;99(5):1848–1855. doi: 10.1083/jcb.99.5.1848. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Cunningham B. A., Hoffman S., Rutishauser U., Hemperly J. J., Edelman G. M. Molecular topography of the neural cell adhesion molecule N-CAM: surface orientation and location of sialic acid-rich and binding regions. Proc Natl Acad Sci U S A. 1983 May;80(10):3116–3120. doi: 10.1073/pnas.80.10.3116. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Edelman G. M. Modulation of cell adhesion during induction, histogenesis, and perinatal development of the nervous system. Annu Rev Neurosci. 1984;7:339–377. doi: 10.1146/annurev.ne.07.030184.002011. [DOI] [PubMed] [Google Scholar]
  5. Finne J., Finne U., Deagostini-Bazin H., Goridis C. Occurrence of alpha 2-8 linked polysialosyl units in a neural cell adhesion molecule. Biochem Biophys Res Commun. 1983 Apr 29;112(2):482–487. doi: 10.1016/0006-291x(83)91490-0. [DOI] [PubMed] [Google Scholar]
  6. Finne J., Mäkelä P. H. Cleavage of the polysialosyl units of brain glycoproteins by a bacteriophage endosialidase. Involvement of a long oligosaccharide segment in molecular interactions of polysialic acid. J Biol Chem. 1985 Jan 25;260(2):1265–1270. [PubMed] [Google Scholar]
  7. Flicker P. F., Peltz G., Sheetz M. P., Parham P., Spudich J. A. Site-specific inhibition of myosin-mediated motility in vitro by monoclonal antibodies. J Cell Biol. 1985 Apr;100(4):1024–1030. doi: 10.1083/jcb.100.4.1024. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Gennarini G., Hirn M., Deagostini-Bazin H., Goridis C. Studies on the transmembrane disposition of the neural cell adhesion molecule N-CAM. The use of liposome-inserted radioiodinated N-CAM to study its transbilayer orientation. Eur J Biochem. 1984 Jul 2;142(1):65–73. doi: 10.1111/j.1432-1033.1984.tb08251.x. [DOI] [PubMed] [Google Scholar]
  9. Gennarini G., Rougon G., Deagostini-Bazin H., Hirn M., Goridis C. Studies on the transmembrane disposition of the neural cell adhesion molecule N-CAM. A monoclonal antibody recognizing a cytoplasmic domain and evidence for the presence of phosphoserine residues. Eur J Biochem. 1984 Jul 2;142(1):57–64. doi: 10.1111/j.1432-1033.1984.tb08250.x. [DOI] [PubMed] [Google Scholar]
  10. Glenney J. R., Jr, Glenney P., Weber K. Mapping the fodrin molecule with monoclonal antibodies. A general approach for rod-like multidomain proteins. J Mol Biol. 1983 Jun 25;167(2):275–293. doi: 10.1016/s0022-2836(83)80336-2. [DOI] [PubMed] [Google Scholar]
  11. Goridis C., Hirn M., Santoni M. J., Gennarini G., Deagostini-Bazin H., Jordan B. R., Kiefer M., Steinmetz M. Isolation of mouse N-CAM-related cDNA: detection and cloning using monoclonal antibodies. EMBO J. 1985 Mar;4(3):631–635. doi: 10.1002/j.1460-2075.1985.tb03676.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Hemperly J. J., Murray B. A., Edelman G. M., Cunningham B. A. Sequence of a cDNA clone encoding the polysialic acid-rich and cytoplasmic domains of the neural cell adhesion molecule N-CAM. Proc Natl Acad Sci U S A. 1986 May;83(9):3037–3041. doi: 10.1073/pnas.83.9.3037. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. 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]
  14. Hoffman S., Sorkin B. C., White P. C., Brackenbury R., Mailhammer R., Rutishauser U., Cunningham B. A., Edelman G. M. Chemical characterization of a neural cell adhesion molecule purified from embryonic brain membranes. J Biol Chem. 1982 Jul 10;257(13):7720–7729. [PubMed] [Google Scholar]
  15. Ibsen S., Berezin V., Nørgaard-Pedersen B., Bock E. Enzyme-linked immunosorbent assay of the D2-glycoprotein. J Neurochem. 1983 Aug;41(2):356–362. doi: 10.1111/j.1471-4159.1983.tb04750.x. [DOI] [PubMed] [Google Scholar]
  16. Kiehart D. P., Kaiser D. A., Pollard T. D. Monoclonal antibodies demonstrate limited structural homology between myosin isozymes from Acanthamoeba. J Cell Biol. 1984 Sep;99(3):1002–1014. doi: 10.1083/jcb.99.3.1002. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Murray B. A., Hemperly J. J., Gallin W. J., MacGregor J. S., Edelman G. M., Cunningham B. A. Isolation of cDNA clones for the chicken neural cell adhesion molecule (N-CAM). Proc Natl Acad Sci U S A. 1984 Sep;81(17):5584–5588. doi: 10.1073/pnas.81.17.5584. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Murray B. A., Hemperly J. J., Prediger E. A., Edelman G. M., Cunningham B. A. Alternatively spliced mRNAs code for different polypeptide chains of the chicken neural cell adhesion molecule (N-CAM). J Cell Biol. 1986 Jan;102(1):189–193. doi: 10.1083/jcb.102.1.189. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Rothbard J. B., Brackenbury R., Cunningham B. A., Edelman G. M. Differences in the carbohydrate structures of neural cell-adhesion molecules from adult and embryonic chicken brains. J Biol Chem. 1982 Sep 25;257(18):11064–11069. [PubMed] [Google Scholar]
  20. Rougon G., Deagostini-Bazin H., Hirn M., Goridis C. Tissue- and developmental stage-specific forms of a neural cell surface antigen linked to differences in glycosylation of a common polypeptide. EMBO J. 1982;1(10):1239–1244. doi: 10.1002/j.1460-2075.1982.tb00019.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. 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]
  22. Rutishauser U., Watanabe M., Silver J., Troy F. A., Vimr E. R. Specific alteration of NCAM-mediated cell adhesion by an endoneuraminidase. J Cell Biol. 1985 Nov;101(5 Pt 1):1842–1849. doi: 10.1083/jcb.101.5.1842. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Sorkin B. C., Hoffman S., Edelman G. M., Cunningham B. A. Sulfation and phosphorylation of the neural cell adhesion molecule, N-CAM. Science. 1984 Sep 28;225(4669):1476–1478. doi: 10.1126/science.6474186. [DOI] [PubMed] [Google Scholar]
  24. Watanabe M., Frelinger A. L., 3rd, Rutishauser U. Topography of N-CAM structural and functional determinants. I. Classification of monoclonal antibody epitopes. J Cell Biol. 1986 Nov;103(5):1721–1727. doi: 10.1083/jcb.103.5.1721. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. 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]

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