Abstract
The binding of fluorescein-conjugated Concanavalin A to the cell surface has been studied in normal and transformed cells in interphase and mitosis. Binding to the cell surface was in the form of an incomplete ring of fluorescence, and the binding was inhibited by α-methyl-D-mannopyranoside. All the cells were fluorescent when treated with 25-250 μg/ml of fluorescent Concanavalin A. With 10 μg, the cells were all fluorescent after 30 min of binding, but after 0.5-5 min with 10 μg or 30 min with 1 or 2.5 μg, transformed interphase cells showed a higher percentage of cells with surface fluorescence than did normal interphase cells. Trypsinized normal and transformed interphase cells showed the same fluorescence. Binding with 2.5 μg at 4° instead of at 24°, gave a higher percentage of fluorescent cells with trypsinized than with untrypsinized transformed cells. Mitotic normal cells were similar to transformed interphase cells, whereas mitotic transformed cells were intermediate between normal and transformed interphase cells. The results indicate that the use of low concentrations of fluorescent Concanavalin A can show differences in surface fluorescence between normal and transformed, interphase and mitotic, and trypsinized and untrypsinized cells. It is suggested that the observed differences in fluorescence can be explained by differences in affinity of the lectin binding sites and/or differences in the clustering of sites.
Keywords: lectins, hamster, mouse 3T3 cells, agglutination, mitosis
Full text
PDF
Images in this article
Selected References
These references are in PubMed. This may not be the complete list of references from this article.
- CHANOCK R. M., HAYFLICK L., BARILE M. F. Growth on artificial medium of an agent associated with atypical pneumonia and its identification as a PPLO. Proc Natl Acad Sci U S A. 1962 Jan 15;48:41–49. doi: 10.1073/pnas.48.1.41. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Fox T. O., Sheppard J. R., Burger M. M. Cyclic membrane changes in animal cells: transformed cells permanently display a surface architecture detected in normal cells only during mitosis. Proc Natl Acad Sci U S A. 1971 Jan;68(1):244–247. doi: 10.1073/pnas.68.1.244. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Huberman E., Salzberg S., Sachs L. The in vitro induction of an increase in cell multiplication and cellular life span by the water-soluble carcinogen dimethylnitrosamine. Proc Natl Acad Sci U S A. 1968 Jan;59(1):77–82. doi: 10.1073/pnas.59.1.77. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Inbar M., Ben-Bassat H., Sachs L. A specific metabolic activity on the surface membrane in malignant cell-transformation. Proc Natl Acad Sci U S A. 1971 Nov;68(11):2748–2751. doi: 10.1073/pnas.68.11.2748. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Inbar M., Ben-Bassat H., Sachs L. Inhibition of ascites tumor development by concanavalin A. Int J Cancer. 1972 Jan 15;9(1):143–149. doi: 10.1002/ijc.2910090117. [DOI] [PubMed] [Google Scholar]
- Inbar M., Sachs L. Interaction of the carbohydrate-binding protein concanavalin A with normal and transformed cells. Proc Natl Acad Sci U S A. 1969 Aug;63(4):1418–1425. doi: 10.1073/pnas.63.4.1418. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Inbar M., Sachs L. Structural difference in sites on the surface membrane of normal and transformed cells. Nature. 1969 Aug 16;223(5207):710–712. doi: 10.1038/223710a0. [DOI] [PubMed] [Google Scholar]
- Klein E., Clifford P., Klein G., Hamberger C. A. Further studies on the membrane immunofluorescence reaction of Burkitt lymphoma cells. Int J Cancer. 1967 Jan 15;2(1):27–36. doi: 10.1002/ijc.2910020106. [DOI] [PubMed] [Google Scholar]
- Nicolson G. L., Singer S. J. Ferritin-conjugated plant agglutinins as specific saccharide stains for electron microscopy: application to saccharides bound to cell membranes. Proc Natl Acad Sci U S A. 1971 May;68(5):942–945. doi: 10.1073/pnas.68.5.942. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Shoham J., Inbar M., Sachs L. Differential toxicity on normal and transformed cells in vitro and inhibition of tumour development in vivo by concanavalin A. Nature. 1970 Sep 19;227(5264):1244–1246. doi: 10.1038/2271244a0. [DOI] [PubMed] [Google Scholar]
- Singer S. J., Nicolson G. L. The fluid mosaic model of the structure of cell membranes. Science. 1972 Feb 18;175(4023):720–731. doi: 10.1126/science.175.4023.720. [DOI] [PubMed] [Google Scholar]
- Sumner J. B., Howell S. F. Identification of Hemagglutinin of Jack Bean with Concanavalin A. J Bacteriol. 1936 Aug;32(2):227–237. doi: 10.1128/jb.32.2.227-237.1936. [DOI] [PMC free article] [PubMed] [Google Scholar]