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. 1974 Aug 1;140(2):437–451. doi: 10.1084/jem.140.2.437

SPECIFIC BINDING OF NERVE GROWTH FACTOR (NGF) BY MURINE C 1300 NEUROBLASTOMA CELLS

R Revoltella 1, L Bertouni 1, M Pediconi 1, E Vigneti 1
PMCID: PMC2139591  PMID: 4211021

Abstract

Murine C 1300 neuroblastoma cells bind with high avidity on their membrane surface the nerve growth factor (NGF), a protein capable of inducing differentiation of sympathetic nerve cells. The total binding capacity of NGF by the cells was quantitatively measured by a radioimmunoassay technique, using 125I-labeled NGF. An average number of about 106 molecules of NGF could be bound, at saturation, by each cell with an average relative association constant of about 107 liters/mol. Using synchronized cells, it was found, however, that either the number of molecules of ligand bound or the avidity of the binding interaction between NGF and cells varied depending upon their growth cycle, the maximal-binding occurring during the G1 and early S phase. Binding of [125I]NGF was suppressed by trypsin treatment of the cells, however new receptor sites were rapidly replaced onto the membrane surface within 1–2 h. Cells exposed to 3 M KCl released into the supernate a protein product exhibiting similar high avidity for NGF. Acrylamide gel electrophoresis suggested a restricted molecular heterogeneity of this product, with a major component in the 52,000 mol wt region. Antibodies made specific to this protein were capable, in the absence of the complement, of inhibiting the binding of [125I]NGF by the cells and in the presence of the complement they killed them.

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Selected References

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

  1. Angeletti R. H., Bradshaw R. A., Wade R. D. Subunit structure and amino acid composition of mouse submaxillary gland nerve growth factor. Biochemistry. 1971 Feb 2;10(3):463–469. doi: 10.1021/bi00779a018. [DOI] [PubMed] [Google Scholar]
  2. Banerjee S. P., Snyder S. H., Cuatrecasas P., Greene L. A. Binding of nerve growth factor receptor in sympathetic ganglia. Proc Natl Acad Sci U S A. 1973 Sep;70(9):2519–2523. doi: 10.1073/pnas.70.9.2519. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Bocchini V., Angeletti P. U. The nerve growth factor: purification as a 30,000-molecular-weight protein. Proc Natl Acad Sci U S A. 1969 Oct;64(2):787–794. doi: 10.1073/pnas.64.2.787. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Buell D. N., Fahey J. L. Limited periods of gene expression in immunoglobulin-synthesizing cells. Science. 1969 Jun 27;164(3887):1524–1525. doi: 10.1126/science.164.3887.1524. [DOI] [PubMed] [Google Scholar]
  5. Byars N., Kidson C. Programmed synthesis and export of immunoglobulin by synchronized myeloma cells. Nature. 1970 May 16;226(5246):648–650. doi: 10.1038/226648a0. [DOI] [PubMed] [Google Scholar]
  6. Dales S. Concerning the universality of a microtubule antigen in animal cells. J Cell Biol. 1972 Mar;52(3):748–754. doi: 10.1083/jcb.52.3.748. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. GREENWOOD F. C., HUNTER W. M., GLOVER J. S. THE PREPARATION OF I-131-LABELLED HUMAN GROWTH HORMONE OF HIGH SPECIFIC RADIOACTIVITY. Biochem J. 1963 Oct;89:114–123. doi: 10.1042/bj0890114. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Haskill J. S., Elliott B. E., Kerbel R., Axelrad M. A., Eidinger D. Classification of thymus-derived and marrow-derived lymphocytes by demonstration of their antigen-binding characteristics. J Exp Med. 1972 Jun 1;135(6):1410–1415. doi: 10.1084/jem.135.6.1410. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Levi-Montalcini R., Angeletti P. U. Nerve growth factor. Physiol Rev. 1968 Jul;48(3):534–569. doi: 10.1152/physrev.1968.48.3.534. [DOI] [PubMed] [Google Scholar]
  10. Marchalonis J. J., Cone R. E., Atwell J. L. Isolation and partial characterization of lymphocyte surface immunoglobulins. J Exp Med. 1972 Apr 1;135(4):956–971. doi: 10.1084/jem.135.4.956. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Olmsted J. B., Carlson K., Klebe R., Ruddle F., Rosenbaum J. Isolation of microtubule protein from cultured mouse neuroblastoma cells. Proc Natl Acad Sci U S A. 1970 Jan;65(1):129–136. doi: 10.1073/pnas.65.1.129. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Petersen D. F., Tobey R. A., Anderson E. C. Developmental biology of normal cells: biology and biochemical aspects. Cancer Res. 1968 Sep;28(9):1821–1822. [PubMed] [Google Scholar]
  13. Prescott D. M. Regulation of cell reproduction. Cancer Res. 1968 Sep;28(9):1815–1820. [PubMed] [Google Scholar]
  14. Reisfeld R. A., Pellegrino M. A., Kahan B. D. Salt extraction of soluble HL-A antigens. Science. 1971 Jun 11;172(3988):1134–1136. doi: 10.1126/science.172.3988.1134. [DOI] [PubMed] [Google Scholar]
  15. Revoltella R., Angeletti R. H., Pediconi M., Bertolini L. Radioimmunoassay for the measurement of mass and avidity of anti-nerve growth factor antibodies. J Immunol Methods. 1974 Jan;3(1):67–82. doi: 10.1016/0022-1759(74)90035-0. [DOI] [PubMed] [Google Scholar]
  16. Revoltella R., Bertolini L., Pediconi M. Unmasking of nerve growth factor membrane-specific binding sites in synchronized murine C 1300 neuroblastoma cells. Exp Cell Res. 1974 Mar 30;85(1):89–94. doi: 10.1016/0014-4827(74)90216-x. [DOI] [PubMed] [Google Scholar]
  17. Revoltella R., Martinelli G., Osler A. G. The antigen-binding capacity of mouse lymphocytes. II. Changes during the early immune response. Cell Immunol. 1973 Feb;6(2):215–230. doi: 10.1016/0008-8749(73)90023-3. [DOI] [PubMed] [Google Scholar]
  18. Schubert D., Humphreys S., Baroni C., Cohn M. In vitro differentiation of a mouse neuroblastoma. Proc Natl Acad Sci U S A. 1969 Sep;64(1):316–323. doi: 10.1073/pnas.64.1.316. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Schubert D., Humphreys S., Jacob F., de Vitry F. Induced differentiation of a neuroblastoma. Dev Biol. 1971 Aug;25(4):514–546. doi: 10.1016/0012-1606(71)90004-2. [DOI] [PubMed] [Google Scholar]
  20. Shapiro A. L., Viñuela E., Maizel J. V., Jr Molecular weight estimation of polypeptide chains by electrophoresis in SDS-polyacrylamide gels. Biochem Biophys Res Commun. 1967 Sep 7;28(5):815–820. doi: 10.1016/0006-291x(67)90391-9. [DOI] [PubMed] [Google Scholar]
  21. Shelanski M. L., Gaskin F., Cantor C. R. Microtubule assembly in the absence of added nucleotides. Proc Natl Acad Sci U S A. 1973 Mar;70(3):765–768. doi: 10.1073/pnas.70.3.765. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Tobey R. A., Ley K. D. Isoleucine-mediated regulation of genome repliction in various mammalian cell lines. Cancer Res. 1971 Jan;31(1):46–51. [PubMed] [Google Scholar]
  23. Yamada K. M., Wessells N. K. Axon elongation. Effect of nerve growth factor on microtubule protein. Exp Cell Res. 1971 Jun;66(2):346–352. doi: 10.1016/0014-4827(71)90687-2. [DOI] [PubMed] [Google Scholar]

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