Skip to main content
NCBI home page
Proceedings of the National Academy of Sciences of the United States of America logoLink to Proceedings of the National Academy of Sciences of the United States of America
. 1973 Jun;70(6):1672–1676. doi: 10.1073/pnas.70.6.1672

Cell Contact-Dependent Increase in Membrane D-Galactopyranosyl-Like Residues on Normal, But Not Virus- or Spontaneously-Transformed, Murine Fibroblasts

Garth L Nicolson 1, Monique Lacorbiere 1
PMCID: PMC433570  PMID: 4352648

Abstract

Ricinus communis agglutinin (a lectin specific for β-D-galactopyranosyl-like oligosaccharide residues) was used to investigate differences in lectin-binding sites on normal 3T3 and transformed SV3T3 and 3T12 cells grown to various cell densities in culture. The agglutinability of SV3T3 cells by R. communis agglutinin decreased when cells were grown to confluency or to a point where the cells were in contact. The agglutinability of 3T3 cells also decreased at confluency, but this result was variable and not as dramatic as with SV3T3 cells. Saturation-binding experiments performed at 4° with 125I-labeled R. communis agglutinin were used to monitor the number of lectin-binding sites on cells grown to different densities. Transformed cells (SV3T3 and 3T12) did not show cell-culture density-dependent changes in R. communis sites; sparse, touching, and confluent transformed cells had equivalent numbers of sites. However, touching (or confluent) 3T3 cells possessed 2.5-times the number of R. communis sites per cell compared to growing, sparsely populated 3T3 cells. When growing sparse 3T3 cells are compared to growing SV3T3 or 3T12 cells, there are 5-times R. communis sites per unit surface area on the transformed cells. The change in R. communis sites during cell growth in culture is discussed in relation to membrane fluidity and topography of plasma membrane oligosaccharides and glycosyl transferases, which can modify cell surfaces at contact.

Keywords: 3T3, SV3T3, and 3T12 cells; 125I-labeled Ricinus communis agglutinin; lectins; cell agglutination; plasma membrane oligosaccharides

Full text

PDF
1672

Images in this article

1674Image
on p.1674

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. Aaronson S. A., Todaro G. J. Basis for the acquisition of malignant potential by mouse cells cultivated in vitro. Science. 1968 Nov 29;162(3857):1024–1026. doi: 10.1126/science.162.3857.1024. [DOI] [PubMed] [Google Scholar]
  3. Aaronson S. A., Todaro G. J. Development of 3T3-like lines from Balb-c mouse embryo cultures: transformation susceptibility to SV40. J Cell Physiol. 1968 Oct;72(2):141–148. doi: 10.1002/jcp.1040720208. [DOI] [PubMed] [Google Scholar]
  4. Agrawal B. B., Goldstein I. J. Protein-carbohydrate interaction. VI. Isolation of concanavalin A by specific adsorption on cross-linked dextran gels. Biochim Biophys Acta. 1967 Oct 23;147(2):262–271. [PubMed] [Google Scholar]
  5. 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]
  6. Bretton R., Wicker R., Bernhard W. Ultrastructural localization of concanavalin A receptors in normal and SV 40 -transformed hamster and rat cells. Int J Cancer. 1972 Sep 15;10(2):397–410. doi: 10.1002/ijc.2910100222. [DOI] [PubMed] [Google Scholar]
  7. Burger M. M. A difference in the architecture of the surface membrane of normal and virally transformed cells. Proc Natl Acad Sci U S A. 1969 Mar;62(3):994–1001. doi: 10.1073/pnas.62.3.994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. 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]
  9. Burger M. M., Martin G. S. Agglutination of cells transformed by Rous sarcoma virus by wheat germ agglutinin and concanavalin A. Nat New Biol. 1972 May 3;237(70):9–12. doi: 10.1038/newbio237009a0. [DOI] [PubMed] [Google Scholar]
  10. 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]
  11. Castor L. N. Flattening, movement and control of division of epithelial-like cells. J Cell Physiol. 1970 Feb;75(1):57–64. doi: 10.1002/jcp.1040750107. [DOI] [PubMed] [Google Scholar]
  12. Ceccarini C., Eagle H. pH as a determinant of cellular growth and contact inhibition. Proc Natl Acad Sci U S A. 1971 Jan;68(1):229–233. doi: 10.1073/pnas.68.1.229. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Clarke G. D., Stoker M. G., Ludlow A., Thornton M. Requirement of serum for DNA synthesis in BHK 21 cells: effects of density, suspension and virus transformation. Nature. 1970 Aug 22;227(5260):798–801. doi: 10.1038/227798a0. [DOI] [PubMed] [Google Scholar]
  14. 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]
  15. Cuatrecasas P. Protein purification by affinity chromatography. Derivatizations of agarose and polyacrylamide beads. J Biol Chem. 1970 Jun;245(12):3059–3065. [PubMed] [Google Scholar]
  16. Drysdale R. G., Herrick P. R., Franks D. The specificity of the haemagglutinin of the Castor bean, Ricinus communis. Vox Sang. 1968;15(3):194–202. doi: 10.1111/j.1423-0410.1968.tb01749.x. [DOI] [PubMed] [Google Scholar]
  17. Dulbecco R., Stoker M. G. Conditions determining initiation of DNA synthesis in 3T3 cells. Proc Natl Acad Sci U S A. 1970 May;66(1):204–210. doi: 10.1073/pnas.66.1.204. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Dulbecco R. Topoinhibition and serum requirement of transformed and untransformed cells. Nature. 1970 Aug 22;227(5260):802–806. doi: 10.1038/227802a0. [DOI] [PubMed] [Google Scholar]
  19. GOLDSTEIN I. J., HOLLERMAN C. E., SMITH E. E. PROTEIN-CARBOHYDRATE INTERACTION. II. INHIBITION STUDIES ON THE INTERACTION OF CONCANAVALIN A WITH POLYSACCHARIDES. Biochemistry. 1965 May;4:876–883. doi: 10.1021/bi00881a013. [DOI] [PubMed] [Google Scholar]
  20. Goto M., Kataoka Y., Goto K., Yokoyama T., Sato H. Decrease in agglutinability of cultured tumor cells to concanavalin-A at the plateau of cell growth. Gan. 1972 Aug;63(4):505–506. [PubMed] [Google Scholar]
  21. Griffiths J. B. Role of serum, insulin and amino acid concentration in contact inhibition of growth of human cells in culture. Exp Cell Res. 1972 Nov;75(1):47–56. doi: 10.1016/0014-4827(72)90518-6. [DOI] [PubMed] [Google Scholar]
  22. Hakomori S. Cell density-dependent changes of glycolipid concentrations in fibroblasts, and loss of this response in virus-transformed cells. Proc Natl Acad Sci U S A. 1970 Dec;67(4):1741–1747. doi: 10.1073/pnas.67.4.1741. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Hakomori S., Kijimoto S. Forssman reactivity and cell contacts in cultured hamster cells. Nat New Biol. 1972 Sep 20;239(90):87–88. doi: 10.1038/newbio239087a0. [DOI] [PubMed] [Google Scholar]
  24. Henkart P., Humphreys T. Cell aggregation in small volumes on a gyratory shaker. Exp Cell Res. 1970 Nov;63(1):224–227. doi: 10.1016/0014-4827(70)90358-7. [DOI] [PubMed] [Google Scholar]
  25. Holley R. W., Kiernan J. A. "Contact inhibition" of cell division in 3T3 cells. Proc Natl Acad Sci U S A. 1968 May;60(1):300–304. doi: 10.1073/pnas.60.1.300. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. 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]
  27. Kijimoto S., Hakomori S. -I. Contact-dependent enhancement of net synthesis of forssman glycolipid antigen and hematoside in nil cells at the early stage of cell-to-cell contact. FEBS Lett. 1972 Sep 1;25(1):38–42. doi: 10.1016/0014-5793(72)80448-4. [DOI] [PubMed] [Google Scholar]
  28. LeVine D., Kaplan M. J., Greenaway P. J. The purification and characterization of wheat-germ agglutinin. Biochem J. 1972 Oct;129(4):847–856. doi: 10.1042/bj1290847. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Lipton A., Klinger I., Paul D., Holley R. W. Migration of mouse 3T3 fibroblasts in response to a serum factor. Proc Natl Acad Sci U S A. 1971 Nov;68(11):2799–2801. doi: 10.1073/pnas.68.11.2799. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Macieira-Coelho A., Avrameas S. Modulation of cell behavior in vitro by the substratum in fibroblastic and leukemic mouse cell lines. Proc Natl Acad Sci U S A. 1972 Sep;69(9):2469–2473. doi: 10.1073/pnas.69.9.2469. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Macieira-Coelho A., Avrameas S. Variations in cell overlapping and growth depending on the substratum. Proc Soc Exp Biol Med. 1972 Apr;139(4):1374–1378. doi: 10.3181/00379727-139-36366. [DOI] [PubMed] [Google Scholar]
  32. Martinez-Palomo A., Wicker R., Bernhard W. Ultrastructural detection of concanavalin surface receptors in normal and in polyoma-transformed cells. Int J Cancer. 1972 May 15;9(3):676–684. doi: 10.1002/ijc.2910090326. [DOI] [PubMed] [Google Scholar]
  33. McFARLANE A. S. Efficient trace-labelling of proteins with iodine. Nature. 1958 Jul 5;182(4627):53–53. doi: 10.1038/182053a0. [DOI] [PubMed] [Google Scholar]
  34. Nicolson G. L., Blaustein J. The interaction of Ricinus communis agglutinin with normal and tumor cell surfaces. Biochim Biophys Acta. 1972 May 9;266(2):543–547. doi: 10.1016/0005-2736(72)90109-5. [DOI] [PubMed] [Google Scholar]
  35. 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]
  36. Noonan K. D., Renger H. C., Basilico C., Burger M. M. Surface changes in temperature-sensitive Simian virus 40-transformed cells. Proc Natl Acad Sci U S A. 1973 Feb;70(2):347–349. doi: 10.1073/pnas.70.2.347. [DOI] [PMC free article] [PubMed] [Google Scholar]
  37. 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]
  38. Pohjanpelto P., Raina A. Identification of a growth factor produced by human fibroblasts in vitro as putrescine. Nat New Biol. 1972 Feb 23;235(60):247–249. doi: 10.1038/newbio235247a0. [DOI] [PubMed] [Google Scholar]
  39. Robbins P. W., Macpherson I. A. Glycolipid synthesis in normal and transformed animal cells. Proc R Soc Lond B Biol Sci. 1971 Feb 16;177(1046):49–58. doi: 10.1098/rspb.1971.0014. [DOI] [PubMed] [Google Scholar]
  40. Roth S., White D. Intercellular contact and cell-surface galactosyl transferase activity (cell culture-mouse-radioautography-contact inhibition-cis-and trans-galactosylation). Proc Natl Acad Sci U S A. 1972 Feb;69(2):485–489. doi: 10.1073/pnas.69.2.485. [DOI] [PMC free article] [PubMed] [Google Scholar]
  41. Rubin H. pH and population density in the regulation of animal cell multiplication. J Cell Biol. 1971 Dec;51(3):686–702. doi: 10.1083/jcb.51.3.686. [DOI] [PMC free article] [PubMed] [Google Scholar]
  42. Sefton B. M., Rubin H. Release from density dependent growth inhibition by proteolytic enzymes. Nature. 1970 Aug 22;227(5260):843–845. doi: 10.1038/227843a0. [DOI] [PubMed] [Google Scholar]
  43. Sela B. A., Lis H., Sharon N., Sachs L. Quantitation of N-acetyl-D-galactosamine-like sites on the surface membrane of normal and transformed mammalian cells. Biochim Biophys Acta. 1971 Dec 3;249(2):564–568. doi: 10.1016/0005-2736(71)90132-5. [DOI] [PubMed] [Google Scholar]
  44. 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]
  45. Sivak A., Ray F., Van Duuren B. L. Phorbol ester tumor-promoting agents and membrane stability. Cancer Res. 1969 Mar;29(3):624–630. [PubMed] [Google Scholar]
  46. Stoker M. G., Rubin H. Density dependent inhibition of cell growth in culture. Nature. 1967 Jul 8;215(5097):171–172. doi: 10.1038/215171a0. [DOI] [PubMed] [Google Scholar]
  47. Stoker M. G., Rubin H. Density dependent inhibition of cell growth in culture. Nature. 1967 Jul 8;215(5097):171–172. doi: 10.1038/215171a0. [DOI] [PubMed] [Google Scholar]
  48. Todaro G. J., Lazar G. K., Green H. The initiation of cell division in a contact-inhibited mammalian cell line. J Cell Physiol. 1965 Dec;66(3):325–333. doi: 10.1002/jcp.1030660310. [DOI] [PubMed] [Google Scholar]
  49. Tomita M., Kurokawa T., Onozaki K., Ichiki N., Osawa T., Ukita T. Purification of galactose-binding phytoagglutinins and phytotoxin by affinity column chromatography using sepharose. Experientia. 1972 Jan 15;28(1):84–85. doi: 10.1007/BF01928278. [DOI] [PubMed] [Google Scholar]
  50. Unanue E. R., Perkins W. D., Karnovsky M. J. Ligand-induced movement of lymphocyte membrane macromolecules. I. Analysis by immunofluorescence and ultrastructural radioautography. J Exp Med. 1972 Oct 1;136(4):885–906. doi: 10.1084/jem.136.4.885. [DOI] [PMC free article] [PubMed] [Google Scholar]
  51. VOGT M., DULBECCO R. Steps in the neoplastic transformation of hamster embryo cells by polyoma virus. Proc Natl Acad Sci U S A. 1963 Feb 15;49:171–179. doi: 10.1073/pnas.49.2.171. [DOI] [PMC free article] [PubMed] [Google Scholar]
  52. Vaheri A., Rucoslahti E., Nordling S. Neuraminidase stimulates division and sugar uptake in density-inhibited cell cultures. Nat New Biol. 1972 Aug 16;238(85):211–212. doi: 10.1038/newbio238211a0. [DOI] [PubMed] [Google Scholar]
  53. Vasiliev J. M., Gelfand I. M., Guelstein V. I., Fetisova E. K. Stimulation of DNA synthesis in cultures of mouse embryo fibroblast-like cells. J Cell Physiol. 1970 Jun;75(3):305–313. doi: 10.1002/jcp.1040750307. [DOI] [PubMed] [Google Scholar]

Articles from Proceedings of the National Academy of Sciences of the United States of America are provided here courtesy of National Academy of Sciences

RESOURCES