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Selective changes in the phosphorylation of endogenous proteins in subcellular fractions from cyclic amp-induced differentiated neuroblastoma cells

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Abstract

The endogenous phosphorylation of specific proteins was studied in subcellular fractions from proliferating and cAMP-induced differentiated neuroblastoma cells. Fractions containing nuclear, membrane-bound, and cytosolic proteins were incubated with [γ-32P]ATP, in the presence and absence of added cyclic nucleotides. Phosphate incorporation into specific proteins was determined by slab-gel electrophoresis of sodium dodecyl sulfate-solubilized reaction products. Cytosol fractions from differentiated cells demonstrated a twofold increase in cAMP-dependent phosphorylation of a specific protein with apparent mol wt of 59,000 daltons and a comparable decrease in cAMP-independent phosphorylation of another protein (97,000). The nuclear fraction of differentiated cells showed an increase in the cAMP-independent phosphorylation of two nonhistone proteins (110,000 and 102,000). Membrane fractions from differentiated cells exhibited a differential decrease in endogenous phosphorylation of specific proteins. Selective alterations in the phosphorylation of specific proteins in various subcellular components may be important biochemical events associated with the increased levels of differentiated functions in neuroblastoma cells in culture.

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References

  1. Hamprecht, B., Traber, J., andHamprecht, E., 1974. Dopamine beta-hydroxylase activity in cholinergic neuroblastoma x glioma hybrid cells; increase of activity by dibutyryl cyclic AMP. FEBS Lett. 42:221–226.

    Google Scholar 

  2. Furmanski, P., Silverman, D. J., andLubins, M., 1971. Expression of differentiated functions in mouse neuroblastoma mediated by dibutyryl cyclic adenosine monophosphate. Nature (London), New Biol. 232:413–415.

    Google Scholar 

  3. Macintyre, E. H., Perkins, J. P., Wintersgil, C. J., andVatter, A. W., 1971. The response in culture of human tumor astrocytes and neuroblasts to N6O′ 2-dibutyryl adenosine 3′,5′-cyclic monophosphoric acid. J. Cell Sci., 11:639–667.

    Google Scholar 

  4. Richelson, E., 1973. Stimulation of tyrosine hydroxylase activity in an adrenergic clone of mouse neuroblastoma by dibutyryl cyclic AMP. Nature (London), New Biol. 242:175–177.

    Google Scholar 

  5. Prasad, K. N., 1975. Differentiation of neuroblastoma cells in culture. Biol. Rev. 50:129–165.

    Google Scholar 

  6. Prasad, K. N., Sinha, P. K., Sahu, S. K., andBrown, J. L., 1975. Binding of cyclic nucleotides with soluble proteins increased in differentiated neuroblastoma cells. Biochem Biophys. Res. Commun. 66:131–138.

    Google Scholar 

  7. Prasad, K. N., Sinha, P. K., Sahu, S. K., andBrown, J. L., 1976. Binding of cyclic nucleotides with proteins in malignant and cAMP-induced differentiated neuroblastoma cells. Cancer Res. 36:2290–2296.

    Google Scholar 

  8. Prashad, N., Wischmeyer, B., Evetts, C., Baskin, F., andRosenberg, R., 1977. Dibutyryl cAMP-induced protein changes in differentiating mouse neuroblastoma cells. Cell Differ. 6:147–157.

    Google Scholar 

  9. Ehrlich, Y. H., Brunngraber, E. G., Sinha, P. K., andPrasad, K. N., 1977. Specific alterations in phosphorylation of cytosol proteins from differentiating neuroblastoma cells. Nature (London) 265:238–240.

    Google Scholar 

  10. Prasad, K. N., andHsie, A. W., 1971. Morphological differentiation of mouse neuroblastoma cells induced in vitro by dibutyryl adenosine 3′∶5′-cyclic monophosphate. Nature (London), New Biol. 233:141–142.

    Google Scholar 

  11. Prasad, K. N., Mandal, B., Waymire, J. C., Lees, G. J., Vernadakis, A., andWeiner, N., 1973. Basal levels of neurotransmitter synthesizing enzymes and effect of cyclic AMP agents on the morphological differentiation of isolated neuroblastoma clones. Nature (London), New Biol. 241:117–119.

    Google Scholar 

  12. Sheppard, H., Wiggan, G., andTsien, W. H., 1972. Structure-activity relationships for inhibitors of phosphodiesterase from erythrocytes and other tissues. Pages 102–112,in Greengard, P., Robinson, G. A., and Paolette, R. (eds.), Advances in Cyclic Nucleotide Research, Raven Press, New York.

    Google Scholar 

  13. Bondy, S. C., Prasad, K. N., andPurdy, J. L., 1974. Neuroblastoma: Drug-induced differentiation increases proportion of cytoplasmic RNA that contains polyadenylic acid. Science 186:359–361.

    Google Scholar 

  14. Ehrlich, Y. H., andRouttenberg, A., 1974. Cyclic AMP regulates phosphorylation of three proteins of rat cerebral cortex membranes for thirty minutes. FEBS Lett. 45:237–243.

    Google Scholar 

  15. Ehrlich, Y. H., Davis, L. G., Gilfoil, T., andBrunngraber, E. G., 1977. Distribution of endogenously phosphorylated proteins in subcellular fractions of rat cerebral cortex. Neurochem. Res. 2:533–548.

    Google Scholar 

  16. Routtenberg, A., andEhrlich, Y. H., 1975. Endogenous phosphorylation of four proteins in cerebral cortex membranes. Role of cyclic nucleotides, ATP and divalent cations. Brain Res. 92:415–430.

    Google Scholar 

  17. Lowry, O. H., Rosebrough, N. J., Farr, A. L., andRandall, R. J., 1951. Protein measurement with the Folin phenol reagent. J. Biol. Chem. 93:265–275.

    Google Scholar 

  18. Ueda, T., Maeno, H., andGreengard, P., 1973. Regulation of endogenous phosphorylation of specific proteins in synaptic membrane fractions from rat brain by adenosine 3′,5′-monophosphate. J. Biol. Chem. 248:8295–8305.

    Google Scholar 

  19. Ehrlich, Y. H., Bonnet, K. A., Davis, L. G., andBrunngraber, E. G., 1977. Protein phosphorylation in neostriatal membranes: Involvement of specific phosphoproteins in narcotic addiction. Pages 273–278,in Lajtha, A., Roberts, S., and Gypsen, W. (eds.), Developments in Neuroscience, Vol. 2, Elsevier, Amsterdam.

    Google Scholar 

  20. Prasad, K. N., Fogleman, D., Gaschier, M., Sinha, P. K., andBrown, J. L., 1975. Cyclic nucleotide-dependent protein kinase activity in malignant and cyclic AMP-induced differentiated neuroblastoma cells in culture. Biochem. Biophys. Res. Commun. 68:1248–1255.

    Google Scholar 

  21. Dutton, G. R., andMahler, H. R., 1968. In vitro RNA synthesis by intact rat brain nuclei. J. Neurochem. 15:765–780.

    Google Scholar 

  22. Hollenbeck, R. A., Chuang, D. M., andCosta, E., 1978. Protein kinase activation and translocation in neuroblastoma. Trans. Am. Soc. Neurochem. 9:75 (abstract).

    Google Scholar 

  23. Ehrlich, Y. H., Davis, L. G., andBrunngraber, E. G., 1978. Effects of decapitation stress on protein phosphorylation in cortical membranes. Brain Res. Bull. 3:251–256.

    Google Scholar 

  24. Ehrlich, Y. H., Rabjohns, R. R., andRouttenberg, A., 1977. Experiential input alters the phosphorylation of specific proteins in brain membranes. Biochem. Pharmacol. Behav. 6:169–174.

    Google Scholar 

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Authors and Affiliations

  1. Biochemistry Laboratories, Missouri Institute of Psychiatry, University of Missouri-Columbia, School of Medicine, 5400 Arsenal Street, 63139, St. Louis, Missouri

    Y. H. Ehrlich, L. G. Davis & E. G. Brunngraber

  2. Department of Radiology, University of Colorado Medical Center, 80262, Denver, Colorado

    K. N. Prasad & P. K. Sinha

Authors
  1. Y. H. Ehrlich
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  2. K. N. Prasad
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  3. P. K. Sinha
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  4. L. G. Davis
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  5. E. G. Brunngraber
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Ehrlich, Y.H., Prasad, K.N., Sinha, P.K. et al. Selective changes in the phosphorylation of endogenous proteins in subcellular fractions from cyclic amp-induced differentiated neuroblastoma cells. Neurochem Res 3, 803–813 (1978). https://doi.org/10.1007/BF00966002

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  • DOI: https://doi.org/10.1007/BF00966002

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