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. 1973 Jun;70(6):1625–1629. doi: 10.1073/pnas.70.6.1625

A Comparative Evaluation of the Distribution of Concanavalin A-Binding Sites on the Surfaces of Normal, Virally-Transformed, and Protease-Treated Fibroblasts

J Z Rosenblith *, T E Ukena †, H H Yin †, R D Berlin †, M J Karnovsky *
PMCID: PMC433559  PMID: 4352644

Abstract

The topographical distributions of concanavalin A-binding sites on the surfaces of 3T3, proteasetreated 3T3, and simian virus 40-transformed 3T3 cultured mouse fibroblasts appear to be different, as shown by a shadow-cast replica technique using concanavalin A and a hemocyanin marker, or as shown previously on isolated membranes with concanavalin A coupled to ferritin. However, chemical fixation of cells before labeling with concanavalin A and hemocyanin, or labeling exclusively at 4°, allows one to distinguish between inherent concanavalin A-binding-site topography and potential rearrangement of sites induced by the action of the multivalent concanavalin A molecule itself. The inherent distribution of binding sites on 3T3, protease-treated 3T3, and transformed cells is actually the same on all cells, i.e., dispersed and random. Treatment of unfixed transformed or protease-treated 3T3 cells, but not normal 3T3 cells, with concanavalin A and hemocyanin at 37° (or at 4° with subsequent warming to 37°), however, results in clustering of binding sites, presumably due to crosslinking of neighboring lectin-binding sites by the quadrivalent concanavalin A. Thus, the underlying difference between concanavalin A-binding sites on normal as compared with transformed or protease-treated normal cells lies not in the inherent topography of binding sites, but rather in the susceptibility of the sites to aggregation by concanavalin A. The latter may reflect an increased mobility of lectin-binding sites on transformed or protease-treated cells.

Keywords: mouse fibroblasts, hemocyanin simian virus 40

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. AUB J. C., TIESLAU C., LANKESTER A. REACTIONS OF NORMAL AND TUMOR CELL SURFACES TO ENZYMES. I. WHEAT-GERM LIPASE AND ASSOCIATED MUCOPOLYSACCHARIDES. Proc Natl Acad Sci U S A. 1963 Oct;50:613–619. doi: 10.1073/pnas.50.4.613. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Arndt-Jovin D. J., Berg P. Quantitative binding of 125 I-concanavalin A to normal and transformed cells. J Virol. 1971 Nov;8(5):716–721. doi: 10.1128/jvi.8.5.716-721.1971. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Burger M. M., Goldberg A. R. Identification of a tumor-specific determinant on neoplastic cell surfaces. Proc Natl Acad Sci U S A. 1967 Feb;57(2):359–366. doi: 10.1073/pnas.57.2.359. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Burger M. M. Proteolytic enzymes initiating cell division and escape from contact inhibition of growth. Nature. 1970 Jul 11;227(5254):170–171. doi: 10.1038/227170a0. [DOI] [PubMed] [Google Scholar]
  5. Cline M. J., Livingston D. C. Binding of 3 H-concanavalin A by normal and transformed cells. Nat New Biol. 1971 Aug 4;232(31):155–156. doi: 10.1038/newbio232155a0. [DOI] [PubMed] [Google Scholar]
  6. Edelman G. M., Cunningham B. A., Reeke G. N., Jr, Becker J. W., Waxdal M. J., Wang J. L. The covalent and three-dimensional structure of concanavalin A. Proc Natl Acad Sci U S A. 1972 Sep;69(9):2580–2584. doi: 10.1073/pnas.69.9.2580. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. HELMKAMP R. W., GOODLAND R. L., BALE W. F., SPAR I. L., MUTSCHLER L. E. High specific activity iodination of gamma-globulin with iodine-131 monochloride. Cancer Res. 1960 Nov;20:1495–1500. [PubMed] [Google Scholar]
  8. 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]
  9. 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]
  10. Kalb A. J., Lustig A. The molecular weight of concanavalin A. Biochim Biophys Acta. 1968 Oct 21;168(2):366–367. doi: 10.1016/0005-2795(68)90161-x. [DOI] [PubMed] [Google Scholar]
  11. Karnovsky M. J., Unanue E. R., Leventhal M. Ligand-induced movement of lymphocyte membrane macromolecules. II. Mapping of surface moieties. J Exp Med. 1972 Oct 1;136(4):907–930. doi: 10.1084/jem.136.4.907. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. LOWRY O. H., ROSEBROUGH N. J., FARR A. L., RANDALL R. J. Protein measurement with the Folin phenol reagent. J Biol Chem. 1951 Nov;193(1):265–275. [PubMed] [Google Scholar]
  13. 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]
  14. Nicolson G. L. Topography of membrane concanavalin A sites modified by proteolysis. Nat New Biol. 1972 Oct 18;239(94):193–197. doi: 10.1038/newbio239193a0. [DOI] [PubMed] [Google Scholar]
  15. Ozanne B., Sambrook J. Binding of radioactively labelled concanavalin A and wheat germ agglutinin to normal and virus-transformed cells. Nat New Biol. 1971 Aug 4;232(31):156–160. doi: 10.1038/newbio232156a0. [DOI] [PubMed] [Google Scholar]
  16. Pinto da Silva P. Translational mobility of the membrane intercalated particles of human erythrocyte ghosts. pH-dependent, reversible aggregation. J Cell Biol. 1972 Jun;53(3):777–787. doi: 10.1083/jcb.53.3.777. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Sharon N., Lis H. Lectins: cell-agglutinating and sugar-specific proteins. Science. 1972 Sep 15;177(4053):949–959. doi: 10.1126/science.177.4053.949. [DOI] [PubMed] [Google Scholar]
  18. 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]
  19. Smith S. B., Revel J. P. Mapping of concanavalin A binding sites on the surface of several cell types. Dev Biol. 1972 Mar;27(3):434–441. doi: 10.1016/0012-1606(72)90183-2. [DOI] [PubMed] [Google Scholar]

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