Molecular and Cellular Biochemistry

, Volume 55, Issue 2, pp 183–190 | Cite as

Grafting of triggering site onto lymphocytes; distribution of grafted dinitrophenyl groups on cell surface glycoproteins and glycolipids

  • Sarah Spiegel
  • Meir Wilchek


Quantitative investigation of membrane-bound sialoglycoconjugates on lymphocyte surface was performed by chemical modification of the sialic acid residues with radioactive N4-dinitrophenyl-l-2,4-diaminobutyric acid hydrazide (DNP-DABH). This labeled both glycoproteins and glycolipids with concomitant preservation of the mitogenic activity by multivalent hapten binding protein (anti-DNP antibody). Under conditions where maximum stimulation of thymocytes occurred radioactive DNP-DABH labeled 1.1 × 107 glycolipids molecules/cell but, only 3 × 106 glycoproteins molecules/cell. When B lymphocytes, which do not undergo DNP-mediated stimulation were used, glycolipids labeling could not be detected. Major differences between stimulation committed and non-committed DNP-modified lymphocytes was the amount of ligand attached to the cell surface sialoglycolipids (gangliosides).


Cell Surface Sialic Acid Chemical Modification Hydrazide Surface Glycoprotein 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.





N4-dinitrophenyl-l-2,4-diaminobutyric acid hydrazide


phytohemagglutinin from the red kidney beanPhaseolus vulgaris

Con A

concanavalin A


phosphate buffered saline, pH 7.2




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  1. 1.
    Post, G., Kirsh, R. and Fidler, I. J., 1979. Cell. Immunol. 44: 71–88.Google Scholar
  2. 2.
    Higgins, I. J., Sabatino, A. P., Remold, H. G. and David, J. R., 1978. J. Immunol. 121: 880–886.Google Scholar
  3. 3.
    Besançon, F. and Ankel, H., 1974. Nature 252: 478–480.Google Scholar
  4. 4.
    Ankel, H., Krishnamurti, C., Besançon, F., Stefanos, S. and Falcoff, E., 1980. Proc. Natl. Acad. Sci. U.S.A. 77: 2528 -2532.Google Scholar
  5. 5.
    O'Brien, K. J., Condie, R. M., Prody, C. A. and Edstrom, R. D., 1979. J. Biol. Chem. 254: 168–174.Google Scholar
  6. 6.
    Benacerraf, B. and Unanue, E. R., 1979. Immunology, Chapter 5, pp. 76–108, Williams and Wilkins, Baltimore/London.Google Scholar
  7. 7.
    Rauvala, H. and Finne, J., 1979. FEBS Lett. 97: 1–8.Google Scholar
  8. 8.
    Wilchek, M., Ravid, A. and Spiegel, S., 1979. Molecular Mechanisms of Biological Recognition (Balaban, M., ed.), pp. 299–309, Elsevier/North-Holland Biomedical Press, Amsterdam.Google Scholar
  9. 9.
    Ravid, A., Novogrodsky, A. and Wilchek, M., 1978. Eur. J. Immunol. 8: 289–294.Google Scholar
  10. 10.
    Wilchek, M. and Givol, D., 1977. Methods Enzymol. 46: 501–504.Google Scholar
  11. 11.
    Haas, E., Wilchek, M., Katchalski-Katzir, E. and Steinberg, I. Z., 1975. Proc. Natl. Acad. Sci. U.S.A. 72: 1807–1811.Google Scholar
  12. 12.
    Reisner, Y., Ravid, A. and Sharon, N., 1976. Biochem. Biophys. Res. Commun. 72: 1585–1591.Google Scholar
  13. 13.
    Greenwood, F. C., Hunter, H. H. and Glover, J. S., 1963. Biochem. J. 89: 114–123.Google Scholar
  14. 14.
    Spiegel, S., Ravid, A. and Wilchek, M., 1979. Proc. Natl. Acad. Sci. U.S.A. 76: 5277–5281.Google Scholar
  15. 15.
    Yates, A. J., Mattison, S. L. and Whisler, R. L., 1980. Biochem. Biophys. Res. Commun. 96: 211–218.Google Scholar
  16. 16.
    Stobo, J. D., Rosenthal, A. S. and Paul, W. E., 1972. J. Immunol. 108: 1–17.Google Scholar
  17. 17.
    Greaves, M. F., Bauminger, S. and Janossy, G., 1972. Clin. Exptl. Immunol. 10: 537–554.Google Scholar
  18. 18.
    Greaves, M. F. and Bauminger, S., 1972. Nature New Biol. 235: 67–70.Google Scholar
  19. 19.
    Andersson, J., Edelman, G. M., Möller, G. and Sjöberg, O., 1972. Eur. J. Immunol. 2: 233–235.Google Scholar
  20. 20.
    Merritt, W. D. and Jelsema, C. L., 1980. Eur. J. Cell. Biol. 22: 274.Google Scholar
  21. 21.
    Rosenfelder, G. and Muhlradt, P. E. C., 1978. Immunitaetsforsch. 155: 54.Google Scholar

Copyright information

© Martinus Nijhoff Publishers 1983

Authors and Affiliations

  • Sarah Spiegel
    • 1
  • Meir Wilchek
    • 1
  1. 1.Department of BiophysicsThe Weizmann Institute of ScienceRehovotIsrael

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