Abstract
Homocystinuria is an inborn error of sulfur amino acid metabolism characterized predominantly by vascular and nervous system dysfunction. In this study we determined the in vitro effects of homocysteine and methionine, metabolites which accumulate in homocystinuria, on Na+, K+-ATPase, and Mg2+-ATPase activities in synaptic membranes from the hippocampus of rats. The results showed that both metabolites significantly inhibit Na+, K+-ATPase but not Mg2+-ATPase activity at concentrations usually observed in plasma of homocystinuric patients. Furthermore, incubation of hippocampal homogenates with homocysteine also elicited an inhibition of the enzyme activity which was however prevented by the simultaneous addition of cysteine to the medium. In addition, cysteine or methionine per se did not modify the two enzymatic activities. These findings indicate that oxidation of critical groups in the enzyme may possibly be involved in homocysteine inhibitory effect. Moreover, kinetic studies performed to investigate the interaction between homocysteine and methionine on Na+, K+-ATPase inhibition suggested a common site for the two amino acids in the enzyme. Considering the critical role exerted by Na+, K+-ATPase in brain, it is proposed that the inhibition provoked by homocysteine and methionine on the enzyme activity may be possibly related to the brain dysfunction characteristic of homocystinuria.
Similar content being viewed by others
REFERENCES
Allen, I.C., Grieve, A., and Griffiths, R. (1986). Differential changes in the content of amino acid neurotransmitters in discrete regions of the rat brain prior to the onset and during the course of homocysteine-induced seizures. J. Neurochem. 46:1582–1592.
Bradford, M.M. (1976). A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye-binding. Anal. Biochem. 72:248–254.
Chan, K.M., Delfer, D., and Junger, K.D. (1986). A direct colorimetric assay for Ca2+-stimulated ATPase activity. Anal. Biochem. 157:375–380.
Chevillard, C., Cárdenas, M.L., and Cornish-Bowden, A. (1993). The competition plot: A simple test of whether two reactions occur at the same active site. Biochem. J. 289:599–604.
Ericinska, M., and Silver, I.A. (1994). Silver, ions and energy in mammalian brain. Prog. Neurobiol. 16:37–71.
Gaull, G.E., Bender, A.N., Vulovic, D., Tallan, H.H., and Schaffner, F. (1980). Methioninemia and myopathy: A new disorder. Ann. Neurol. 9:423–432.
Grisar, T. (1984). Glial and neuronal Na+-K+ pump in epilepsy. Ann. Neurol. 16(Suppl.):128–134.
Hattori, H., and Kanfer, J.N. (1984). Inhibition of rat brain microsomal Na+,K+-ATPase by S-adenosylmethionine. J. Neurochem. 42:204–208.
Henneberry, R.L., Novelli, A., Cox, J.A., and Lysko, P.G. (1989). Neurotoxicity at the N-methyl-D-aspartate receptor in energy-compromised neurons. An hypothesis for cell death in aging and disease. Ann. NY Acad. Sci. 568:225–233.
Hogg, N. (1999). The effect of cyst(e)ine on the auto-oxidation of homocysteine. Free Radic. Biol. Med. 27:28–33.
Jones, D.H., and Matus, A.I. (1974). Isolation of plasma synaptic membrane from brain by combination flotationsedimentation density gradient centrifugation. Biochim. Biophys. Acta 356:276–287.
Kim, J.P., Koh, J.-Y., and Choi, D.W. (1987). L-homocysteate is a potent neurotoxin on cultured cortical neurons. Brain Res. 437:103–110.
Kim, W.-K., and Pae, Y.-S. (1996). Involvement of N-Methyl-D-aspartate receptor and free radical in homocysteine-mediated toxicity on rat cerebellar granule cells in culture. Neurosci. Lett. 216:117–120.
Kubová, H., Folbergrová, J., and Mares, P. (1995). Seizures induced by homocysteine in rats during ontogenesis. Epilepsia 36:750–756.
Labrune, P., Perignon, J.L., Rault, M., Brunet, C., Lutun, H., Charpentier, C., Sdubray, J.M., and Odievre, M. (1990). Familial hypermethioninemia partially responsive to dietary restriction. J. Pediatr. 117:220–226.
Lees, G.J. (1993). Contributory mechanisms in the causation of neurodegenerative disorders. Neuroscience 54:287–322.
Malinow, M.R. (1990). Hyperhomocyst(e)inemia. A common and easily reversible risk factor for occlusive atherosclerosis. Circulation 81:2004–2006.
McIlwain, H., and Poll, J.D. (1985). Interation between adenosine generated endogenously in neocortical tissues, and homocysteine and its thiolactone. Neurochem. Int. 7:103–105.
Mudd, S.H., Levy, H.L., and Skovby, F. (2001). Disorders of transsulfuration. In (C.R. Scriver, A.L. Beaudet, W.S. Sly, and D. Valle, eds.), The Metabolic and Molecular Bases of Inherited Disease, 8th ed., McGraw-Hill, New York, pp. 1279–1327.
Olney, J.W., Prince, M.T., Salles, K.S., Labruyere, J., Ryerson, R., Mahan, K., Frierdich, G., and Samson, L. (1987). L-Homocysteic acid: An endogenous excitotoxic ligand of the NMDA receptor. Brain Res. Bull. 19:597–601.
Pontes, Z.E.L., Oliveira, L.S., Bavaresco, C.S., Streck, E.L., Dutra-Filho, C.S., Wannmacher, C.M.D., and Wyse, A.T.S. (1999). Proline administration decreases Na, K-ATPase activity in the synaptic plasma membrane from cerebral cortex of rats. Metab. Brain Dis. 14:265–273.
Renkawek, K., Renier, W.O., De Pont, J.J., Vogels, O.J., and Gabreels, F.J. (1992). Neonatal status convulsivus, spongiform encephalopathy, and low activity of Na+/K(+)-ATPase in the brain. Epilepsy 33:58–64.
Silva, C.G., Parolo, E., Streck, E.L., Wajner, M., Wannmacher, C.M.D., and Wyse, A.T.S. (1999). In vitro inhibition of Na+, K+-ATPase activity from cerebral cortex by guanidino compounds accumulating in hyperargininemia. Brain Res. 838:78–84.
Streck, E.L., Edom, P.T., Noriler, M.E., Borges, L.F., Pontes, Z.L., Parolo, E., Dutra-Filho, C.S., Wannmacher, C.M.D, and Wyse, A.T.S. (2000). Effect of phenylalanine and p-chlorophenylalanine on Na+,K+-ATPase activity in the synaptic plasma membrane from the cerebral cortex of rats. Metab. Brain Dis. 15:105–114.
Tsakiris, S., and Deliconstantinos, G. (1984). Influence of phosphatidylserine on (Na++ K+)-stimulated ATPase and acetylcholinesterase activities of dog brain synaptossomal plasma membranes. Biochem. J. 22:301–307.
Tsakiris, S., Angelogianni, P., Schulpis, K.H., and Behrakis, P. (2000). Protective effect of L-cysteine and glutathione on rat brain Na+, K+-ATPase inhibition induced by free radicals. Z. Naturforsch. 55:271–277.
Van der Hijden, H.T., Schuurmans Stekhoven, F.M., and De Pont, J.J. (1989). Sidedness of the effect of amines on the steady-state phosphorylation level of reconstituted Na+/K+-ATPase. Biochim. Biophys. Acta 987:75–82.
Welch, G.N., Upchurch, G.R., and Loscalzo, J. (1997). Homocysteine, oxidative stress, and vascular disease. Hosp. Pract. 32:81–92.
Wyse, A.T.S., Wajner, M., and Wannmacher, C.M.D. (1998). Kinetics of alanine reversal on the inhibition of Na+, K+-ATPase activity by phenylalanine and phenyllactate in the synaptic plasma membrane from the cerebral cortex of rats. Med. Sci. Res. 26:141–143.
Wyse, A.T.S., Noriler, M.E., Borges, L.F., Floriano, P.J., Silva, C.G., Wajner, M., and Wannmacher, C.M.D. (1999). Alanine prevents the decrease of Na+, K+-ATPase activity in experimental phenylketonuria. Metab. Brain Dis. 14:95–101.
Wyse, A.T.S., Streck, E.L., Worm, P., Wajner, A., Ritter, F., and Netto, C.A. (2000). Preconditioning prevents the inhibition of Na+, K+-ATPase activity after brain ischemia. Neurochem. Res. 25:969–973.
Xu, K.Y., Zweier, J.L., and Becker, L.C. (1997). Oxygen-free radicals directly attack he ATP binding site of the cardiac Na+, K+-ATPase. Ann. NY Acad. Sci. 834:680–683.
Yufu, K., Itoh, T., Edamatsu, R., Mori, A., and Hirakawa, M. (1993). Effect of hyperbaric oxygenation on the Na+, K(+)-ATPase and membrane fluidity of cerebrocortical membranes after experimental subarachnoid hemorrhage. Neurochem. Res. 16:1033–1039.
Author information
Authors and Affiliations
Corresponding author
Rights and permissions
About this article
Cite this article
Streck, E.L., Zugno, A.I., Tagliari, B. et al. Inhibition of Na+, K+-ATPase Activity by the Metabolites Accumulating in Homocystinuria. Metab Brain Dis 17, 83–91 (2002). https://doi.org/10.1023/A:1015594111778
Issue Date:
DOI: https://doi.org/10.1023/A:1015594111778