Abstract
12 distinct neural cell adhesion molecule (N-CAM) epitopes, each recognized by a different monoclonal antibody (mAb), have been characterized in terms of the major structural and functional features of the molecule. Seven antibodies, each recognizing the amino-terminal region of the molecule, altered the rate of N-CAM-mediated adhesion. Four of these were inhibitors, two of which also recognized a heparin- binding N-CAM fragment. The other three antibodies specifically enhanced the rate of N-CAM-mediated adhesion. Three epitopes, one polypeptide- and two carbohydrate-dependent, were associated with the sialic acid-rich central portion of the molecule. The remaining two antibodies were found to react with intracellular determinants, and are specific for the largest of the three major N-CAM polypeptide forms. Studies on the ability of one antibody to hinder recognition of native N-CAM by another antibody suggested that the epitopes associated with N- CAM binding functions are in close proximity compared with the other determinants. The classification of these mAb epitopes has allowed the topographical placement of key N-CAM features, as described in the following paper, and provides valuable probes for analysis of both the structure and function of N-CAM.
Full Text
The Full Text of this article is available as a PDF (1.4 MB).
Selected References
These references are in PubMed. This may not be the complete list of references from this article.
- Brodeur B. R., Tsang P., Larose Y. Parameters affecting ascites tumour formation in mice and monoclonal antibody production. J Immunol Methods. 1984 Jul 6;71(2):265–272. doi: 10.1016/0022-1759(84)90073-5. [DOI] [PubMed] [Google Scholar]
- Buskirk D. R., Thiery J. P., Rutishauser U., Edelman G. M. Antibodies to a neural cell adhesion molecule disrupt histogenesis in cultured chick retinae. Nature. 1980 Jun 12;285(5765):488–489. doi: 10.1038/285488a0. [DOI] [PubMed] [Google Scholar]
- 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]
- Cole G. J., Loewy A., Glaser L. Neuronal cell-cell adhesion depends on interactions of N-CAM with heparin-like molecules. Nature. 1986 Apr 3;320(6061):445–447. doi: 10.1038/320445a0. [DOI] [PubMed] [Google Scholar]
- 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]
- Elder J. H., Alexander S. endo-beta-N-acetylglucosaminidase F: endoglycosidase from Flavobacterium meningosepticum that cleaves both high-mannose and complex glycoproteins. Proc Natl Acad Sci U S A. 1982 Aug;79(15):4540–4544. doi: 10.1073/pnas.79.15.4540. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 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]
- 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]
- Frelinger A. L., 3rd, Rutishauser U. Topography of N-CAM structural and functional determinants. II. Placement of monoclonal antibody epitopes. J Cell Biol. 1986 Nov;103(5):1729–1737. doi: 10.1083/jcb.103.5.1729. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 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]
- 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]
- Hahn H. J., Hellman B., Lernmark A., Sehlin J., Täljedal I. B. The pancreatic beta-cell recognition of insulin secretogogues. Influence of neuraminidase treatment on the release of insulin and the islet content of insulin, sialic acid, and cyclic adenosine 3':5'-monophosphate. J Biol Chem. 1974 Aug 25;249(16):5275–5284. [PubMed] [Google Scholar]
- Hall A. K., Rutishauser U. Phylogeny of a neural cell adhesion molecule. Dev Biol. 1985 Jul;110(1):39–46. doi: 10.1016/0012-1606(85)90061-2. [DOI] [PubMed] [Google Scholar]
- Hawkes R., Niday E., Gordon J. A dot-immunobinding assay for monoclonal and other antibodies. Anal Biochem. 1982 Jan 1;119(1):142–147. doi: 10.1016/0003-2697(82)90677-7. [DOI] [PubMed] [Google Scholar]
- 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]
- 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]
- 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]
- Lemmon V., Staros E. B., Perry H. E., Gottlieb D. I. A monoclonal antibody which binds to the surface of chick brain cells and myotubes: cell selectivity and properties of the antigen. Brain Res. 1982 Mar;255(3):349–360. doi: 10.1016/0165-3806(82)90003-7. [DOI] [PubMed] [Google Scholar]
- Mage M. G. Preparation of Fab fragments from IgGs of different animal species. Methods Enzymol. 1980;70(A):142–150. doi: 10.1016/s0076-6879(80)70045-9. [DOI] [PubMed] [Google Scholar]
- Oakley B. R., Kirsch D. R., Morris N. R. A simplified ultrasensitive silver stain for detecting proteins in polyacrylamide gels. Anal Biochem. 1980 Jul 1;105(2):361–363. doi: 10.1016/0003-2697(80)90470-4. [DOI] [PubMed] [Google Scholar]
- Rutishauser U., Grumet M., Edelman G. M. Neural cell adhesion molecule mediates initial interactions between spinal cord neurons and muscle cells in culture. J Cell Biol. 1983 Jul;97(1):145–152. doi: 10.1083/jcb.97.1.145. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 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]
- Rutishauser U., Thiery J. P., Brackenbury R., Edelman G. M. Adhesion among neural cells of the chick embryo. III. Relationship of the surface molecule CAM to cell adhesion and the development of histotypic patterns. J Cell Biol. 1978 Nov;79(2 Pt 1):371–381. doi: 10.1083/jcb.79.2.371. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 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]
- 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]
- Swanstrom R., Shank P. R. X-Ray Intensifying Screens Greatly Enhance the Detection by Autoradiography of the Radioactive Isotopes 32P and 125I. Anal Biochem. 1978 May;86(1):184–192. doi: 10.1016/0003-2697(78)90333-0. [DOI] [PubMed] [Google Scholar]
- Thanos S., Bonhoeffer F., Rutishauser U. Fiber-fiber interaction and tectal cues influence the development of the chicken retinotectal projection. Proc Natl Acad Sci U S A. 1984 Mar;81(6):1906–1910. doi: 10.1073/pnas.81.6.1906. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Tosney K. W., Watanabe M., Landmesser L., Rutishauser U. The distribution of NCAM in the chick hindlimb during axon outgrowth and synaptogenesis. Dev Biol. 1986 Apr;114(2):437–452. doi: 10.1016/0012-1606(86)90208-3. [DOI] [PubMed] [Google Scholar]
- Towbin H., Staehelin T., Gordon J. Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications. Proc Natl Acad Sci U S A. 1979 Sep;76(9):4350–4354. doi: 10.1073/pnas.76.9.4350. [DOI] [PMC free article] [PubMed] [Google Scholar]
- WARREN L. Thiobarbituric acid spray reaction for deoxy sugars and sialic acids. Nature. 1960 Apr 16;186:237–237. doi: 10.1038/186237a0. [DOI] [PubMed] [Google Scholar]