Abstract
Endogenous phosphorylation of proteins from rat brain synaptosomal plasma membranes was studied in vitro. Cyclic AMP (cAMP) markedly stimulated32P incorporation in three protein bands with molecular weights of 75,000, 57,000, and 54,000, respectively. The effect of the behaviorally active peptide ACTH1–24 on this endogenous phosphorylation in vitro was studied using peptide concentrations from 10−10 to 10−4 M. In a number of protein bands, a biphasic effect of ACTH1–24 was observed: in concentrations of 10−4–10−5 M, a reduced amount of32P was found; in concentrations of 10−6–10−7 M, hardly any effect could be detected, whereas consistently at concentrations around 10−8 M, a significant decrease was again observed. The phosphoprotein bands affected by in vitro addition of ACTH1–24 were of a smaller molecular weight than those affected by in vitro addition of cAMP.
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Walton, K. G., DeLorenzo, R. J., Curran, P. F., andGreengard, P. 1975. Regulation of protein phosphorylation and sodium transport in toad bladder. J. Gen. Phys. 65:153–177.
Greengard, P. 1976. Possible role for cyclic nucleotides and phosphorylated membrane proteins in postsynaptic actions of neurotransmitters. Nature (London) 260:101–108.
Glassman, E., Gispen, W. H., Perumal, R., Machlus, B., andWilson, J. E. 1972. The effect of short experiences on the incorporation of phosphate into synaptosomal and non-histone acid extractable nuclear proteins from rat and mouse. 5th International Congress on Pharmacology, San Francisco, July 23–28th, p. 265 (abstract).
Perumal, R., Gispen, W. H., Wilson, J. E., andGlassman, E. 1975, Phosphorylation of proteins from the brains of mice subjected to short-term behavioral experiences. Pages 201–207,in Progress in Brain Research,Gispen, W. H., Van Wimersma Greidanus, Tj. B., Bohus, B., andDe Wied, D. (eds.), Vol. 42, Elsevier Publishing Co., Amsterdam.
Routtenberg, A., Ehrlich, Y. H., andRabjohns, R. R. 1975. Effect of a training experience of phosphorylation of a specific protein in neocortical and subcortical membrane preparations. Fed. Proc. Fed. Am. Soc. Exp. Biol. 34:17.
De Wied, D. 1974. Pituitary-adrenal system hormones and behavior. Pages 653–666,in The Neurosciences, Third Study Program,Schmitt, F. O., andWorden, F. G. (eds.), Rockefeller University Press, New York.
Schotman, P., Reith, M. E. A., Van Wimersma Greidanus, Tj. B., Gispen, W. H., andDe Wied, D. 1976. Hypothalamic and pituitary peptide hormones and the central nervous system: with special reference to the neurochemical effects of ACTH. Pages 309–344,in Molecular and Functional Neurobiology,Gispen, W. H. (ed.), Elsevier Publishing Co., Amsterdam.
Terenius, L. 1973. Stereospecific interaction between narcotic analgesics and a synaptic plasma membrane fraction of rat cerebral cortex. Acta Pharmacol. Toxicol. 32:317.
Routtenberg, A., andEhrlich, Y. H. 1975. Endogenous phosphorylation of four cerebral cortical membrane proteins: Role of cyclic nucleotides, ATP and divalent cations. Brain Res. 92:415–430.
Lowry, O. H., Rosebrough, N. J., Farr, A. L., andRandall, R. J. 1951. Protein measurement with the Folin phenol reagent. J. Biol. Chem. 193:265–275.
Lugtenberg, B., Meijers, J., Peters, R., van der Hoek, P., andvan Alphen, L. 1975. Electrophoretic resolution of the major outer membrane protein ofEscherichia coli K12 into four bands. FEBS Lett. 58:254–258.
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.
Ehrlich, Y. H., andRouttenberg, A. 1974. Cyclic AMP regulates phosphorylation of three protein components of rat cerebral cortex membranes for thirty minutes. FEBS Lett. 45(1):237–243.
Schotman, P., Gispen, W. H., Jansz, H. S., andDe Wied, D. 1972. Effects of ACTH analogues on macromolecule metabolism in the brain stem of hypophysectomized rats. Brain Res. 46:349–362.
Reith, M. E. A., Schotman, P., andGispen, W. H. 1974. Hypophysectomy, ACTH1–10 and in vitro protein synthesis in rat brain stem slices. Brain Res. 81:571–575.
Reith, M. E. A., Schotman, P., andGispen, W. H. 1975. Incorporation of [3H]leucine into brain stem protein fraction: The effect of a behaviorally active,N-terminal fragment of ACTH in hypophysectomized rats. Neurobiology 5:355–368.
Reith, M. E. A., Schotman, P., andGispen, W. H. 1975. The neurotropic action of ACTH: Effects of ACTH-like peptides on the incorporation of leucine into protein of brain stem slices from hypophysectomized rats. Neurosci. Lett. 1:55–59.
Rudman, D., Scott, J. W., Del Rio, A. E., Houser, D. H., andSheen, S. 1974. Effect of melanotropic peptides on protein synthesis in mouse brain. Am. J. Physiol. 226:687–692.
Rees, H. D., Brogan, L. L., Entingh, D. J., Dunn, A. J., Shinkman, P. G., Damstra-Entingh, T., Wilson, J. E., andGlassman, E. 1974. Effect of sensory stimulation on the uptake and incorporation of radioactive lysine into protein of mouse brain and liver. Brain Res. 68:143–156.
Versteeg, D. H. G. 1973. Effect of two ACTH analogues on noradrenaline metabolism in rat brain. Brain Res. 49:483–485.
Wiegant, V. M., andGispen, W. H. 1975. Behaviorally active ACTH analogs and brain cyclic AMP. Exp. Brain Res. 23(Suppl.):219.
Rudman, D., andIsaacs, J. W. 1975. Effect of intrathecal injection of melanotropiclipolytic peptides on the concentration of 3′,5′-cyclic adenosine monophosphate in cerebrospinal fluid. Endocrinology 97:1476–1480.
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Zwiers, H., Veldhuis, H.D., Schotman, P. et al. ACTH, cyclic nucleotides, and brain protein phosphorylation in vitro. Neurochem Res 1, 669–677 (1976). https://doi.org/10.1007/BF00965607
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DOI: https://doi.org/10.1007/BF00965607