Skip to main content
SpringerLink
Log in

Effect of phenylalanine and its metabolites on ATP diphosphohydrolase activity in synaptosomes from rat cerebral cortex

  • Original Articles
  • Published:
Neurochemical Research Aims and scope Submit manuscript

Abstract

The in vitro effects of phenylalanine and some of its metabolites on ATP diphosphohydrolase (apyrase, EC 3.6.1.5) activity in synaptosomes from rat cerebral cortex were investigated. The enzyme activity in synaptosomes from rats subjected to experimental hyperphenylalaninemia (α-methylphenylalanine plus phenylalanine) was also studied. In the in vitro studies, a biphasic effect of phenylalanine on both enzyme substrates (ATP and ADP) was observed, with maximal inhibition at 2.0 mM and maximal activation at 5.0 mM. Inhibition of the enzyme activity was not due to calcium chelation. Moreover, phenylpyruvate, when compared with phenylalanine showed opposite effects on the enzyme activity, suggesting that phenylalanine and phenylpyruvate bind to two different sites on the enzyme. The other tested phenylalanine metabolites (phenyllactate, phenylacetate and phenylethylamine) had no effect on ATP diphosphohydrolase activity. In addition, we found that ATP diphosphohydrolase activity in synaptosomes from cerebral cortex of rats with chemically induced hyperphenylalaninemia was significantly enhanced by acute or chronic treatment. Since it is conceivable that ATPase-ADPase activities play an important role in neurotransmitter (ATP) metabolism, it is tempting to speculate that our results on the deleterious effects of phenylalanine and phenylpyruvate on ATP diphosphohydrolase activity may be related to the neurological dysfunction characteristics of naturally and chemically induced hyperphenylalaninemia.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Similar content being viewed by others

References

  1. Edwards, F. A., Gibb, A. J., and Colquhoun, D. 1992. ATP receptor-mediated synaptic currents in the central nervous system. Nature. 359:144–147.

    PubMed  Google Scholar 

  2. Evans, R. J., Derkach, V., and Suprenant, A. 1992. ATP mediates fast synaptic transmission in mammalian neurons. Nature. 357:503–505.

    PubMed  Google Scholar 

  3. Burnstock, G. 1986. Purines as cotrnsmitters in adrenergic and chollnergic neurones. Progl. Brain Res. 68:193–203.

    Google Scholar 

  4. Zimmermann, H., Dowdall, M. J., and Lane, D. A. 1979. Purine salvage at the cholinergic nerve endings of the Torpedo electric organ: the central role of adenosine. Neuroscience 4:979–983.

    PubMed  Google Scholar 

  5. Silinsky, E. M. 1975. On the association between transmitter secretion and the release of adenine nucleotides from mammalian motor nerve terminals. J. Physiol. (London). 247:145–162.

    Google Scholar 

  6. Kuroda, Y. 1978. Physiological roles of adenosine derivatives which are released during neurotransmission in mammalian brains. J. Physiol. (Paris). 74:463–470.

    Google Scholar 

  7. Richardson, P. J., and Brown, S. J. 1987. ATP release from affinity-purified rat cholinergic nerve terminals. J. Neurochem. 48:622–630.

    PubMed  Google Scholar 

  8. Sorensen, R. G., and Mahler, H. R. 1981. Calcium-stimulatd adenosine triphosphatase in synaptic membranes. J. Neurochem. 37:1407–1418.

    PubMed  Google Scholar 

  9. Nagy, A. K., Shuster, T. A., and Rosenberg, M. D. 1983. Adenosine triphosphate activity at the external surface of chicken brain synaptosomes. J. Neurochem. 40:226–234.

    PubMed  Google Scholar 

  10. Naby, A. K., Shuster, T. A., and Delgado Escueta, A. V. 1986. Ecto-ATPase of mammalian synaptosomes: Identification and enzymatic characterization. J. Neurochem. 47:976–986.

    PubMed  Google Scholar 

  11. Keller, F., and Zimmerman, H. 1983. Ecto-adenosine triphosphatase activity at this cholinergic and endings of the Torpedo marmorata electric organ. Life Science. 33:2635–2641.

    Google Scholar 

  12. Grondal, E. J., and Zimmermann, H. 1986. Ectonucleotidase activities associated with cholinergic synaptosomes isolated from Torpedo marmorata organ. J. Neurochem. 47:871–881.

    PubMed  Google Scholar 

  13. Battastini, A. M. O., Rocha, J. B. T., Barcellos, C. K., and Dias, R. D. 1991. Characterization of ATP diphophohydrolase (EC 3.6, 1.5) in synaptosomes from cerebral cortex of adult rats. Neurochem. Res. 16:1303–1310.

    PubMed  Google Scholar 

  14. Sarkis, J. J. F., and Saltó, C. 1991. Characterization of a synaptosomal ATP diphsophohydroalase from the electric organ of Torpedo mamorata. Brain Res. Bull. 26:871–876.

    PubMed  Google Scholar 

  15. Schadeck, R. J. G., Sarkis, J. J. F., Dias, R. D., Araujo, H. M. M., Souza, D. O. G. 1989. Synaptosomal apyrase in the hypothalmus of adult rats. Braz. J. Med. Biol. Res. 22:303–314.

    PubMed  Google Scholar 

  16. Rocha, J. B. T., Mello, C. F., Sarkis, J. J. F., and Dias, R. D. 1990. Undernutrition during the preweaning period changes calcium ATPase and ADPase activities of synaptosomal fractions of weanling rats. Br. J. Nutr. 63:273–283.

    PubMed  Google Scholar 

  17. Muller, J., Rocha, J. B. T., Battastini, A. M. O., Sarkis, J. J. F., and Dias, R. D. 1993. Postnatal devlopment of ATPase-ADPase activities in synaptosomal fraction from cerebral cortex of rats. Neurochem. int.-in press.

  18. Ewards, D. J., and Blau, K. 1972. Aromatic derived from phenylalamine in tissues of rats with experimentally induced phenylketoruic-like characteristic. Biochem. J. 130:495–503.

    PubMed  Google Scholar 

  19. Loo, Y.-H., Scotto, J., and Wisniewski, H. M. 1978. Myelin deficiency in experimental phenylketonuria: contribution of the aromatic acid metabolites of phenylalanine. Adv. Exp. Med. Biol. 10:453–469.

    Google Scholar 

  20. Zelnicek, E., and Poddhradsku, O. 1969. Clearance of engogenous phenylpyruvate in phenylketonuria. Clin. Chim. Acta 25:174–180.

    PubMed  Google Scholar 

  21. Tourian, A., and Sibury, J. B. 1983. Phenylketonuria and hyperphenylalanimedia. In Stanbury, J. B., Wyngarden, J. B., Frederickson, D. S. J. Inhr. Metab. Dis. 5 ed., New York, McGraw Hill, pp 270–286.

    Google Scholar 

  22. Devalle, J. A., Daniel, G., and Greengard, O. 1978. Comparison of α-methylphenylalanine and ϱ-chlorophenylalanine as inducers of chronic hyperphenylalaninaemia in developing rats. Biochem. J. 170:449–459.

    PubMed  Google Scholar 

  23. Bulfield, G. 1980. Inherited metabolic disease in laboratory animals: a review. J. Inher. Metab. Dis. 3:133–144.

    PubMed  Google Scholar 

  24. Dwivedy, D. J., and Shah, S. 1982. Effects of phenylalanine and deamined metabolites on Ns+, K+-ATPase activity in synaptosomes from rat brain. Eur. Chem. Res. 7:717–723.

    Google Scholar 

  25. Greengard, O., Yoss, M. S., and Delvalle, J. A. 1976. α-methylpehnylalanine, a new inducer of chronic hyperphenylalaninemia in suckling rats. Science. 192:1007–1008.

    PubMed  Google Scholar 

  26. Huether, G., Klaus, R., and Neuroff, V., 1985. Amino acid depletion in the blood and brain tissue of hyperphenylalaninemic rats is abolished by the administration of additional lysine: a contribution to the understanding of the metabolic defects in phenylketornuria. Biochem. Med. 33:334–341.

    PubMed  Google Scholar 

  27. Greengard, O., and Brass, C. A. 1984. Developmental changes of cerebral phenylalanine uptake from severely elevated blood levels. Neurochem. Res. 9:837–848.

    PubMed  Google Scholar 

  28. Rubin, M. A., Wannmacher, C. M. D., Valente, G. B., Camargo, M. M., and Purer, R. P. 1992. Diminished concentration of the NF-H subunit of neurofilaments in cerebral cortex of rats chronically treated with proline, methylmalonate and phenylalanine plus α-methylphenylalanine. J. Inher. Metab. Dis. 15:252–260.

    PubMed  Google Scholar 

  29. Nagy, A. K., and Delgado Escueta, A. V. 1984. Rapid preparation of synaptosomes from mammalian brain using nontoxic isoosmotic gradient material (Percoll). J. Neurochem. 43:1174–1123.

    Google Scholar 

  30. Lanzetta, P. A., Alvarez, L. J., Reinach, P. S., and Candia, O. A. 1979. An improved assay for nanomole amounts of inorganic phosphate. Anal. Blochem. 100:95–97.

    Google Scholar 

  31. Whittaker, J. T. 1969. A general colorimetric procedure for the estimation of enzymes which are linked to the NADH-NAD+ system. Clin. Chim. Acta. 24:23–37.

    PubMed  Google Scholar 

  32. Bradford, M. M. 1976. A rapid and sensitive method for the quantitation of microgram quatities of protein utlizing the principle of protein-die binding. Anal. Biochem. 72:248–254.

    PubMed  Google Scholar 

  33. Ting-Beall, H. P., and Wells, W. W. 1971. Effect of calcium chelation by L-phenylalanine on (Na+−K+)-stimulated ATPase of chick brain microsomes. FEBS Lett. 16:352–354.

    PubMed  Google Scholar 

  34. Langenbeck, V., Behbehani, A., and Mench-Malnowski, A. 1992. A synposis of the unconjugated acidic transaministration metabolites of phenylalanine in phenylketorunia. J. Inher. Metab. Dis 15:136–144.

    PubMed  Google Scholar 

  35. Harb, J., Meflat, K., Duflos, Y., and Bernad, S. 1984. Evidence for a metalloprotein structure of plasma membrane 5′ nucleotidase. FEBS Lett. 171:215–220.

    PubMed  Google Scholar 

  36. Hommes, F. A. 1991. On the mechanism of permanent brain dysfunction in hyperphenylalaninemia. Biochem. Med. Met. Biol. 46:277–287.

    Google Scholar 

  37. Changeux, J. P., and Dauchin, A. 1976. Selective stabilization of developing synapses as a mechanism for the specification of neuronal network. Nature. 264:705–712.

    PubMed  Google Scholar 

  38. Hommes, F. A., and Matsuo, K. 1987. On a possible mechanism of abnormal brain development in experimental hyperphenylalaninemia. Neurochem. Int. 11:1–10.

    Google Scholar 

  39. Nigan, M. P., and Labor, D. R. 1979. The effect of hyperphenylalaninemia on size and density of synapses in rat neocortex. Brain Res. 179:195–198.

    PubMed  Google Scholar 

  40. Bauman, M. L., and Kemper, T. L. 1982. Morphologic and histoanatomic observations of the brain in untreated human phenylketonuria. Acta Neuropath. 58:55–63.

    PubMed  Google Scholar 

  41. Bremmer, J. H., Duran, M., Kamerling, P. J., Przyrembel, H., and Wadman, K. S. 1981. In Disturbances of amino acid metabolism: clinical chemistry and diagnosis. Pages 306–325,in Urban and Schwarzenberg. (eds.) Baltimore-Munich.

Download references

Author information

Authors and Affiliations

  1. Departamento de Clências Fisiológicas, Fundacão Universidade do Rio Grande, Rio Grande, RS, Brazil

    Angela Terezinha De Souza Wyse & Clovis Milton Duval Wannmacher

  2. Departmento de Bloquímica, Instituto de Biociências, Universidade Federal do Rio Grande do Sul, Rua Sarmento Leite, 500, Porto Alegre, RS, Brazil

    João José Freitas Sarkis, João Sabino Cunha-Filho, Marcio Vieira Teixeira, Maria Rosa Schetinger & Moacir Wajner

Authors
  1. Angela Terezinha De Souza Wyse
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. João José Freitas Sarkis
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. João Sabino Cunha-Filho
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Marcio Vieira Teixeira
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Maria Rosa Schetinger
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Moacir Wajner
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Clovis Milton Duval Wannmacher
    View author publications

    You can also search for this author in PubMed Google Scholar

Rights and permissions

Reprints and permissions

About this article

Cite this article

De Souza Wyse, A.T., Freitas Sarkis, J.J., Cunha-Filho, J.S. et al. Effect of phenylalanine and its metabolites on ATP diphosphohydrolase activity in synaptosomes from rat cerebral cortex. Neurochem Res 19, 1175–1180 (1994). https://doi.org/10.1007/BF00965152

Download citation

  • Accepted:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF00965152

Key Words

Search

Navigation