Abstract
In the present work we investigated the in vitro effects of homocysteine (Hcy) and methionine (Met), metabolites accumulated in homocystinuria, on butyrylcholinesterase (BuChE) activity in rat serum. We also studied the kinetics of the inhibition of BuChE activity caused by Hcy. For determination of BuChE we used serum of 60-day-old Wistar rats, which was incubated in the absence (control) or presence of Hcy (0.01–0.5 mM) or Met (0.2–2.0 mM). The kinetics of the interaction of Hcy and BuChE was determined using the Lineweaver–Burk double reciprocal plot. Results showed that serum BuChE activity was not altered by Met, but it was significantly inhibited (37%) by 500 μM Hcy, a concentration similar to those found in blood of homocystinuric patients. The apparent K m values, in the absence and presence of 500 μM of Hcy, were 0.034 and 0.142 mM, respectively, and V max of BuChE for acetylcholine (ACh) as substrate was 1.25 μmol ACSCh/h/mg of protein. The K i value obtained was 120 μM, and the inhibition was of the competitive type, suggesting a common binding site for Hcy and ACh. It is proposed that inhibition of cholinesterase activity may be one of the mechanisms involved in the neurological dysfunction observed in homocystinuria.
This is a preview of subscription content, log in via an institution to check access.
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.
Benzi, G., and Moretti, A. (1998). Is there a ratinale for the use of acetylcholinesterase inhibitors in the therapy of Alzheimer's disease? Eur. J. Pharmacol. 346:1-13.
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.
Chatonnet, A., and Lockridge, O. (1989). Comparison of butyrylcholinesterase and acetylcholinesterase. Biochem. J. 260:625-634.
Cokugras, A.N., and Teczan, F. (1997). Amitriptyline: A potent inhibitor of butyrylcholinesterase from human serum. Gen. Pharmacol. 29:835-838.
Cummings, J.L. (2000). The role of cholinergic agents in the management of behavioral disturbances in Alzheimer's disease. Int. J. Neuropsychopharmacol. 3:21-29.
De Franchis, R., Sperandeo, M.P., Sebastio, G., and Andria, G. (1998). Clinical aspects of cystathionine β-synthase: How wide is the spectrum? Eur. J. Pediatr. 157 (Suppl. 2):S67-S70.
Dubovy, P. (1991). Histochemical study of non-specific cholinesterase in PNS as a tool for the recognition of relationships between axon and Schwann cells. In Cholinesterases, Structure, Function, Mechanism, Genetics, and Cell Biology, p. 122.
Dubovy, P., and Haninec, P. (1990). Non-specific cholinesterase activity of the developing peripheral nerves and its possible function in cells in intimate contact with growing axons of chick embryo. Ont. J. Dev. Neurosci. 8:589-602.
Ekholm, M. (2001). Predicting relative binding free energies of substrate and inhibitors of acetylcholin-and butyrylcholinesterases. J. Mol. Struct. 572:25-34.
Ellman, G.L., Courtney, K.D., Andres, V., and Featherstone, R.M. (1961). A new and rapid colorimetric determination of acetylcholinesterase activity. Biochem. Pharmacol. 7:88-95.
Giacobini, E. (2001). Selective inhibitors of butyrylcholinesterase, a valid alternative for therapy of Alzheimer's disease? Drugs Aging 18:891-898.
Gómez-Ramos, P., and Morán, M.A. (1997). Ultraestructural localization of butyrylcholinesterase in senile plaques in the brains of aged and Alzheimer's disease patients. Mol. Chem. Neuropathol. 30:161-173.
Harel, M., Schalk, I., Ehret-Sabatier, L., Bouet, F., Goeldner, M., Hirth, C., Axelsen, P.H., Silman, I., and Sussman, J.L. (1993). Quaternary ligand binding to aromatic residues in the active-site gorge of acetylcholinesterase. Proc. Natl. Acad. Sci. U.S.A. 90:9031-9035.
Karczmar, A. (1998). Anticholinesterase: Dramatic aspects of their use and misuse. Neurochem. Int. 32:401-411.
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.
Kraus, J.P. (1998). Biochemistry and molecular genetics of cystathionine β-synthase deficiency. Eur. J. Pediatr. 157(Suppl. 2):S50-S53.
Kubová, H., Folbergrová, J., and Mares, P. (1995). Seizures induced by homocysteine in rats during ontogenesis. Epilepsia 36:750-756.
Kuhn, W., Roebroek, R., Blom, H., Van Oppenraaij, D., Przuntek, H., Kretschmer, A., Buttner, T., Woitalla, D., and Muller, T. (1998). Elevated plasma levels of homocysteine in Parkinson's disease. Eur. Neurol. 40:225-227.
Jaganathan, L., and Boopathy, R. (2000). Distinct effect of benzalkonium chloride on the esterase and aryl acylamidase activities of butyrylcholinesterase. Bioorg. Chem. 28:242-251.
Law, A., Gauthier, S., and Quirion, R. (2001). Say NO Alzheimer's disease: The putative links between nitric oxide and dementia of the Alzheimer's type. Brain Res. Rev. 35:73-96.
Leblhuber, F., Walli, J., Artner-Dworzak, E., Vrecko, K., Widner, B., Reibnegger, G., and Fuchs, D. (2000). Hyperhomocysteinemia in dementia. J. Neural. Transm. 107:343-353.
Lineweaver, H., and Burk, D. (1934). The determination of enzyme dissociation constants. J. Am. Chem. Soc. 56:658-666.
Mack, A., and Robitzki, A. (2000). The key role of butyrylcholinesterase during neurogenesis and neural disorders: An antisense-5′-butyrylcholinesterase-DNA study. Prog. Neurobiol. 60:607-628.
Malinow, M.R. (1990). Hyperhomocyst(e)inemia. A common and easily reversible risk factor for occlusive atherosclerosis. Circulation 81:2004-2006.
Massoulié, J., Pezzementi, L., Bom, S., Krejci, E., and Vallete, F.-M. (1993). Molecular and cellular biology of cholinesterases. Prog. Neurobiol. 41:31-91.
Mattson, M.P., Kruman, I.I., and Duan, W. (2002). Folic acid and homocysteine in age-related disease. Ageing Res. Rev. 1:95-111.
Mcllwain, H., and Poll, J.D. (1985). Interaction between adenosine generated endogenously in neocortical tissues, and homocysteine and its thiolactone. Neurochem. Int. 7:103-105.
Mesulam, M.-M., Guillozet, A., Shaw, P., Levey, A., Duysen, E.G., and Lockridge, O. (2002). Acetylcholinesterase knockouts establish central cholinergic pathways and can use butyrylcholinesterase to hydrolyze acetylcholine. Neuroscience 110:627-639.
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, Vol. 2, McGraw-Hill, New York, pp. 1279-1327.
Reis, E.A., Zugno, A.I., Franzon, R., Tagliari, B., Matté, C., Lamers, M.L. Netto, C. A., and Wyse, A.T.S. (2002). Pretreatment with vitamins E and C prevent the impairment of memory caused by homocysteine administration in rats. Metab. Brain Dis. 17:211-217.
Rocha, J.B.T., Emanuelli, T., and Pereira, M.E. (1993). Effects of early undernutrition on kinetic parameters of brain acetylcholinesterase from adult rats. Acta Neurobiol. Exp. 53:431-437.
Seshadri, S., Beiser, A., Selhub, J., Jacques, P.F., Rosenberg, I.H., D'Agostino, R.B., Wilson, P.W., and Wolf, P.A. (2002). Plasma homocysteine as a risk for dementia and Alzheimer's disease. New Engl. J. Med. 346:476-483.
Streck, E.L., Zugno, A.I., Tagliari, B., Franzon, R., Wannmacher, C.M.D., Wajner, M., and Wyse, A.T.S. (2001). Inhibition of rat brain Na+,K+-ATPase activity induced by homocysteine is probably mediated by oxidative stress. Neurochem. Res. 26:1195-1200.
Streck, E.L., Zugno, A.I., Tagliari, B., Wannmacher, C.M.D., Wajner, M., and Wyse, A.T.S. (2002a). Inhibition of Na+,K+-ATPase activity by the metabolites accumulating in homocystinuria. Metab. Brain Dis. 17:83-91.
Streck, E.L., Matté, C., Vieira, P.S., Rombaldi, F., Wannmacher, C.M.D., Wajner, M., and Wyse, A.T.S. (2002b). Reduction of Na+,K+-ATPase activity in hippocampus of rats subjected to chemically induced hyperhomocysteinemia. Neurochem. Res. 27:1593-1598.
Szegletes, T., Mallander, W.D., Thomas, P.J., and Rosenberry, T.L. (1999). Substrate binding to the peripheral site of acetylcholinesterase initiates enzymatic catalysis: Substrate inhibition arises as a secondary effect. Biochemistry 38:122-133.
Temple, M.E., Luzier, A.B., and Kaazierad, D.J. (2000). Homocysteine as a risk factor for atherosclerosis. Ann. Pharmacol. 34:57-65.
Tohgi, H., Abe, T., Kimura, M., Saheki, M., and Takahashi, S. (1996). Cerebrospinal fluid acetylcholine and choline in vascular dementia of Binswanger and multiple small infarct types as compared with Alzheimer-type dementia. J. Neural. Transm. 103:1211-1220.
Tsakiris, S., Angelogianni, P., Schulpis, K.H., and Stavridis, J.C. (2000). Protective effect of L-phenylalanine on rat brain acetylcholinesterase inhibition induced by free radicals. Clin. Biochem. 33:103-106.
White, A.R., Huang, X., Jobling, M.F., Barrow, C.J., Beyreuther, K., Masters, C.L., Bsh, A.I., and Coppai, R. (2001). Homocysteine potentiates copper-and amyloid beta peptide-mediated toxicity in primary neuronal cultures: Possible risk factors in the Alzheimer's-type neurodegenerative pathways. J. Neurochem. 76:1509-1520.
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Stefanello, F.M., Zugno, A.I., Wannmacher, C.M.D. et al. Homocysteine Inhibits Butyrylcholinesterase Activity in Rat Serum. Metab Brain Dis 18, 187–194 (2003). https://doi.org/10.1023/A:1025551031767
Issue Date:
DOI: https://doi.org/10.1023/A:1025551031767