Abstract
The effect of eight different acetylcholinesterase inhibitors (AChEIs) on the activity of acetylcholinesterase (AChE) molecular forms was investigated. Aqueous-soluble and detergent-soluble AChE molecular forms were separated from rat brain homogenate by sucrose density sedimentation. The bulk of soluble AChE corresponds to globular tetrameric (G4), and monomeric (G1) forms. Heptylphysostigmine (HEP) and diisopropylfluorophosphate were more selective for the G1 than for the G4 form in aqueous-soluble extract. Neostigmine showed slightly more selectivity for the G1 form both in aqueous- and detergent-soluble extracts. Other drugs such as physostigmine, echothiophate, BW284C51, tetrahydroaminoacridine, and metrifonate inhibited both aqueous- and detergent-soluble AChE molecular forms with similar potency. Inhibition of aqueous-soluble AChE by HEP was highly competitive with Triton X-100 in a gradient, indicating that HEP may bind to a detergent-sensitive non-catalytic site of AChE. These results suggest a differential sensitivity among AChE molecular forms to inhibition by drugs through an allosteric mechanism. The application of these properties in developing AChEIs for treatment of Alzheimer disease is considered.
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Massouliè, J., and Bon, S. 1982. The molecular forms of cholinesterase and acetylcholinesterase in vertebrates. Ann. Rev. Neurosci. 5:57–106.
Brimijoin, S. 1983. Molecular forms of acetylcholinesterase in brain, nerve and muscle: nature, localization and dynamics. Prog. Neurobiol. 21:291–322.
Rakonczay, Z. 1986. Mammalian brain acetylcholinesterase. Pages 319–360,in Neuromethods. Neurotransmitter enzymes, Vol. 5, Humana Press Inc., New Jersey.
Inestrosa, N. C., Reiness, C. G., Reichardt, L. F., and Hall, Z. W. 1981. Cellular localization of the molecular forms of acetylcholinesterase in rat pheochromocytoma PC12 cells treated with nerve growth factor. J. Neurosci. 1:1260–1267.
Di Giamberardino, L., and Couraud, J. Y. 1978. Rapid accumulation of high molecular weight acetylcholinesterase in transected sciatic nerve. Nature (London) 271:170–172.
Rieger, F., and Vigny, M. 1976. Solubilization and physicochemical characterization of rat brain acetylcholinesterase: development and maturation of its molecular forms. J. Neurochem. 27:121–129.
Vigny, M., Bon, S., Massouliè, J., and Leterrier, F. 1978. Active-site catalytic efficiency of acetylcholinesterase molecular forms in Electrophorus, Torpedo, rat and chicken. Eur. J. Biochem. 85:317–323.
Bon, S., and Massouliè, J. 1976. An active monomeric form of Electrophorus electricus acetylcholinesterase. FEBS Lett. 67:99–103.
Gentinetta, R., and Brodbeck, U. 1976. Differences in subunit activities in acetylcholinesterase as possible cause for apparent deviation from normal Michaelis-Menten kinetics. Biochem. Biophys. Acta. 438:437–448.
Inestrosa, N. C., Roberts, W. L., Marshall, T. L., and Rosenberry, T. L. 1986. Acetylcholinesterase from bovine caudate nucleus is attached to membranes by a novel subunit distinct from those of acetylcholinesterases in other tissues. J. Biol. Chem. 262:4441–4444.
Andres, C., Mourabit, M., Stutz, C., Mark, J., and Waksman, A. 1990. Are soluble and membrane-bound rat brain acetylcholinesterase different? Neurochem. Res. 15:1065–1072.
Skau, K. A. 1981. Ethopropazine inhibition of AChE molecular forms. Pharmacologist 23:224.
Skau, K. A. 1982. Differential pharmacology of acetylcholinesterase molecular forms. Pharmacologist 24:221.
Volpe, M. T., Bisso, G. M., and Michalek, H. 1990. In vivo and in vitro effects of diisopropylfluorophosphate and Paraxon on individual molecular forms of rat brain acetylcholinesterase. Neurochem. Res. 15:975–979.
Becker, R. E., and Giacobini, E. 1988. Mechanisms of cholinesterase inhibition in senile dementia of the Alzheimer type: clinical, pharmacological and therapeutic aspects. Drug Development. Res. 12:163–195.
Pomponi, M., Giacobini, E., and Brufani, M. 1990. Present state and future development of the therapy of Alzheimer disease. Aging 2:125–153.
Atack, J. R., Perry, E. K., Bonham, J. R., Perry, R. H., Tomlinson, B. E., Blessed, G., and Fairbairn, A. 1983. Molecular forms of acetylcholinesterase in senile dementia of Alzheimer type: selective loss of the intermediate (10 s) form. Neurosci. Lett. 40:199–204.
Fishman, E. B., Siek, G. C., MacCallum, R. D., Bird, E. D., Volicer, L., and Marquis, J. K. 1986. Distribution of the molecular forms of acetylcholinesterase in human brain: alterations in dementia of the Alzheimer type. Ann. Neurol. 19:246–252.
Younkin, S. G., Goodridge, B., Katz, J., Lockett, G., Nafziger, D., Usiak, M. F., and Younkin, L. H. 1986. Molecular forms of acetylcholinesterases in Alzheimer's disease. Federation Proc. 45:2982–2988.
Siek, G. C., Katz, L. S., Fishman, E. B., Koros, T. S., and Marquis, J. K. 1990. Molecular forms of acetylcholinesterase in subcortical areas of normal and Alzheimer disease brain. Biol. Psychiatry 27:573–580.
Nakano, S., Kato, T., Nakamura, S., and Kameyama, M. 1986. Acetylcholinesterase activity in cerebrospinal fluid of patients with Alzheimer's disease and senile dementia. J. Neurol. Sci. 75:213–223.
Bisso, G. M., Diana, G., Fortuna, S., Meneguz, A., and Michalek, H. 1988. Change in the distribution of acetylcholinesterase molecular forms in frontoparietal cortex of the rat following nucleus basalis lesion with kainic acid. Brain Res. 449:391–394.
Brufani, M., Marta, M., and Pomponi, M. 1986. Anticholinesterase activity of a new carbamate, heptyl-physostigmine (C8) in view of its use in patients with Alzheimer-type dementia. Eur. J. Biochem. 157:115–120.
DeSarno, P., Pomponi, M., Giacobini, E., Tang, X. C., and Williams, E. 1989. The effect of heptyl-physostigmine, a new cholinesterase inhibitor, on the central cholinergic system of the rat. Neurochem. Res. 14:971–977.
Johnson, C. D., and Russell, R. L. 1975. A rapid, simple radiometric assay for cholinesterase, suitable for multiple determinations. Analyt. Biochem. 64:229–238.
Grassi, J., Vigny, M., and Massouliè, J. 1982. Molecular forms of acetylcholinesterase in bovine caudate nucleus and superior cervical ganglion: solubility properties and hydrophobic character. J. Neurochem. 38:457–469.
Lazer, M., and Vigny, M. 1980. Modulation of the distribution of acetylcholinesterase molecular forms in a murine neuroblastoma x sympathetic ganglion cell hybrid cell line. J. Neurochem. 35:1067–1079.
Taylor, P. B., Rieger, F., Shelanski, M. L., and Greene, L. A. 1981. Cellular localization of the multiple molecular forms of acetylcholinesterase in cultured neuronal cells. J. Biol. Chem. 256:3827–3830.
Ferrand, C., Clarous, D., Delteil, C., and Weber, M. J. 1986. Cellular localization of the molecular forms of acetylcholinesterase in primary cultures of rat sympathetic neurons and analysis of the secreted enzyme. J. Neurochem. 46:349–358.
Gisinger, V., and Vigny, M. 1977. A specific form of acetylcho-linesterase is secreted by rat sympathetic ganglia. FEBS Lett. 84:253–256.
Changeux, J. P. 1966. Responses of acetylcholinesterase from Torpedo marmorata to salts and curarizing drugs. Mol. Pharmacol. 2:369–392.
Zonetta, J. P., Raconczay, Z., Reeber, A., and Vincedon, G. 1981. Antibodies against the membrane-bound acetylcholinesterase from adult rat brain. FEBS Lett. 129:293–296.
Sørensen, K., Gentinetta, R., and Brodbeck, U. 1982. An amphiphile-dependent form of human brain caudate nucleus acetylcholinesterase: purification and properties. J. Neurochem. 39:1050–1060.
Roufogalis, B. D., and Quist, E. E. 1971. Relative binding sites of pharmacologically active ligand on bovine erythrocyte acetylcholinesterase. Mol. Pharmacol. 8:41–49.
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Special issue dedicated to Dr. Morris H. Aprison.
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Ogane, N., Giacobini, E. & Messamore, E. Preferential inhibition of acetylcholinesterase molecular forms in rat brain. Neurochem Res 17, 489–495 (1992). https://doi.org/10.1007/BF00969897
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DOI: https://doi.org/10.1007/BF00969897