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Acetylcholinesterase inhibition by 1-methyl-4-phenylpyridinium ion, a bioactivated metabolite of MPTP

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Abstract

The effect of the neurotoxicant, 1-methyl-4-phenylpyridinium ion (MPP+) on acetylcholinesterase (AchE) activity was investigated. The MPP+ was found to inactivate the enzyme in a dose dependent manner. The kinetic parameter, Km for the substrate (acetylthiocholine), was found to be 0.216 mM and Ki for MPP+ for the inactivation of AChE was found to be 0.197 mM. It was found that MPP+ is neither a substrate of AChE nor the time-dependent inactivator. The studies of reaction kinetics indicate inactivation of AChE to be a linear mixed-type inhibition. The inactivation of AChE by MPP+ was partially recovered by either dilution or gel exclusion chromatography. These data suggest that once MPP+ enters the basal ganglia of the brain, it can inactivate the AChE and thereby increase the acetylcholine level in the basal ganglia, leading to potential cell dysfunction. It appears likely that the nigrostriatal toxicity by MPP+ leading to Parkinson's disease-like syndrome may, in part, be mediated via the AChE inactivation.

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References

  1. Burnhan WM: Drugs acting on the basal ganglia. In: Principles of Medical Pharmacology. B.C. Decker Inc., Philadelphia, 1989, pp 198–202

    Google Scholar 

  2. Francis GS: Modulation of peripheral sympathetic nerve transmission. J Am Coll Cardiol 12:250–254, 1988

    Google Scholar 

  3. Flattery KV, Spero L: Autonomic nervous system neurotransmitters. In: Principles of Medical Pharmacology. B.C. Decker Inc., Toronto, Philadelphia, 1989, pp 198–202

    Google Scholar 

  4. Ballinger J-C, Levy-Serpier J, Debord J, Penicaut B: Acetylcholinesterase inhibition by two phosphoric 4-nitroanilides. J Enz Inhib 3:211–217, 1990

    Google Scholar 

  5. Fest C, Schmidt KJ: The chemistry of organophosphorus pesticides. Springer Verlag, Berlin-Heidelberg, 1973

    Google Scholar 

  6. Eto M: Organophosphate pesticides: Organic and biological chemistry, Cleveland. The Chemical Rubber Company, 1979

  7. Langston JW, Ballard P, Tetrud JW, Irwin I: Chronic parkinsonism in humans due to a product of meperidine-analog synthesis. Science 219:979–980, 1983

    Google Scholar 

  8. Burns RS, Chiueh CC, Markey SP, Ebert MH, Jacobowitz DM, Kopin IJ: A primate model of parkinsonism: selective destruction of dopaminergic neurons in the pars compactra of the substantia nigra by N-methyl-4-phenyl-1,2,3,6-tetrahydropyridine. Proc Natl Acad Sci USA 80:4546–4550, 1983

    Google Scholar 

  9. Heikkila RE, Manzino L, Cabbat FS, Duvoisin RC: Protection against the dopaminergic neurotoxicity of 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine in mice. Nature (London) 311:467–469, 1984

    Google Scholar 

  10. Langston JW, Irwin I, Langston EB, Forno LS: 1-methyl-4-phenylpyridinium ion (MPP+): Identification of a metabolite of MPTP, a toxin selective to the substantia nigra. Neurosci Lett 48:87–92, 1984

    Google Scholar 

  11. Chiba K, Trevor A, Castagnoli N: Metabolism of the neurotoxic amine, MPTP, by brain monoamine oxidase. Biochem Biophys Res Commun 120:574–579, 1984

    Google Scholar 

  12. Chiba K, Peterson LA, Castagnoli KP, Trevor AJ, Castagnoli NE Jr Studies on the molecular mechanism of bioactivation of the selective nigrostriatal toxin 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine. Drug Metab Dispos 13:342–347, 1985

    Google Scholar 

  13. Gessner W, Bross A, Shen R-S, Abell CW: Further insight into the mode of action of the neurotoxin 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP). FEBS Lett 183:345–348, 1985

    Google Scholar 

  14. Heikkila RE, Cabbat FS, Manzino L, Duvoisin RC: Effect of 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine on nigrostriatal dopamine in mice. Neuropharmacol 23:711–713, 1984

    Google Scholar 

  15. Salach JI, Singer TP, Castagnoli N Jr, Travor A: Oxidation of the neurotoxic amine 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP) by monoamine oxidases A and B and suicide inactivation of the enzymes by MPTP. Biochem Biophys Res Commun 125: 831–825, 1984

    Google Scholar 

  16. Youdim MBH, Finberg PM: New directions in monoamine oxidase A and B selective inhibitors and substrates. Biochem Pharmacol 41:155–162, 1991

    Google Scholar 

  17. Castagnoli N Jr, Sparatore A, Ottoboni S, Leung L, Trevor A: Mechanistic studies on the MPTP inactivation of MAO-B. 10th Intl. Congress of Pharmacology. M.J. Rand and C. Raper (eds), 1987, pp 875–878

  18. Lewin R: Brain enzyme is the target of drug toxin: A chemical known as MPTP causes a Parkinson-like state in humans and monkeys; biochemical and autoradiographical studies are closing in on the mechanism. Science 225:1460–1462, 1984

    Google Scholar 

  19. Riederer P, Jellinger K, Seemann D: In: KF Tipton, P Dostert, M Strolin Benedetti (eds) Monoamine oxidase and disease. Prospects for therapy with reversible inhibitors. Academic Press, New York, 1984, pp 403–415

    Google Scholar 

  20. Ellman GL, Courtney KD, Andres V Jr, Featherstone RM: A new and rapid colorimetric determination of acetylcholinesterase activity. Biochem Pharmacology 7:88–95, 1961

    Google Scholar 

  21. Ellman GL: Tissue sulfhydryl groups. Arch Biochem Biophys 82: 70–77, 1959

    Google Scholar 

  22. Branchini BR, Lajiness E: Inactivation of acetylcholinesterase with a bretylium tosylate photoaffinity probe. Biochim Biophys Acta 884:135–141, 1986

    Google Scholar 

  23. Dixon M, Webb E, Thorne CJR, Tipton KF: Enzyme inhibition and activation. In: Enzymes. Academic Press, New York, 1979, pp 332–467

    Google Scholar 

  24. Ghag S, Wright A, Moudgil VK: Inactivation of rat brain acetylcholinesterase by pyridoxal 5′-phosphate. Biochim Biophys Acta 881:30–37, 1986

    Google Scholar 

  25. Segel IH: Enzyme inhibition. In: Biochemical Calculations. Published by John Wiley & Sons, Inc., 1976, pp 246–272

  26. Wecker L, Laskowski MB, Dettbarn WD: Neuromuscular dysfunction induced by acetylcholinesterase inhibition. Fed Proc 37: 2818–2822, 1978

    Google Scholar 

  27. Kuhn DE, Logan DM, Rathbone MP: Altered acetylcholinesterase isozyme patterns in mice with hereditary muscular dystrophy. J Exp Zool 216:213–233, 1981

    Google Scholar 

  28. Bonham JR, Gowenlock AH, Timothy JA: Acetylcholinesterase and butyrylcholinesterase measurement in the pre-natal detection of neural tube defects and other fetal malformations. Clin Chim Acta 115:163–170, 1981

    Google Scholar 

  29. Soininen H, Halonen T, Riekkinen PJ: Acetylcholinesterase activities in cerebrospinal fluid of patients with senile dementia of Alzheimer type. Acta Neurol Scand 64:217–224, 1981

    Google Scholar 

  30. Ramsay RR, Salach JI, Singer TP: Uptake of the neurotoxin 1-methyl-4-phenylpyridine (MPP+) and its relation to the inhibition of mitochondrial NADH-linked substrates by MPP+. Biochem Biophys Res Commun 134:743–747, 1986

    Google Scholar 

  31. Ramsay RR, Salach JI, Dadgar J, Singer TP: Inhibition of mitochondrial NADH dehydrogenase by pyridine derivatives and its possible relation to idiopathic Parkinsonism. Biochem Biophys Res Commun 135:269–274, 1986

    Google Scholar 

  32. Ramsay RR, McKeowown KA, Johnson EA, Booth RG, Singer TP: Inhibition of NADH oxidation by pyridine derivatives. Biochem Biophys Res Commun 146:53–57, 1987

    Google Scholar 

  33. Ramsay RR, Youngster SK, Nicklas WJ, McKeown KA, Jin Y-Z, Heikkila RE, Singer TP: Structural dependence of the inhibition of mitochondrial respirtion and of NADH oxidase by 1-methyl-4-phenylpyridinium (MPP+) analogs and their energized accumulation by mitochondria. Proc Natl Acad Sci USA 86:9168–9172, 1989

    Google Scholar 

  34. Spinedi A, Pacini L, Limatola C, Luly P, Farias RN: A study of human erythrocyte acetylcholinesterase inhibition by chlorpromazine. Biochem J 278:461–463, 1991

    Google Scholar 

  35. Stoops JK, Bender ML: A spectrophotometric assay for determining the rate constants of acetylcholinesterase inhibitions. Anal Biochem 63:543–554, 1975

    Google Scholar 

  36. Vallette FM, Marsh DJ, Muller F, Massoulie J: Comparative affinity chromatography of acetylcholinesterase from five vertebrate species. J Chromatog 257:285–296, 1983

    Google Scholar 

  37. Edwards JA, Brimijoin S: Thermal inactivation of the molecular forms of acetylcholinesterase and butyrylcholinesterase. Biochim Biophys Acta 742:509–516, 1983

    Google Scholar 

  38. Das YT, Brown HD, Chattopadhyay SK: Microcalorrimetric determination of binding sites of acetylcholinesterase. Biochim Biophys Acta 745:107–110, 1983

    Google Scholar 

  39. Forberg A, Puu G: Kinetics for the inhibition of acetylcholinesterase from the electric eel by some organophosphates and carbamates. J Biochem 140:153–156, 1984

    Google Scholar 

  40. De Jong LPA, Kossen SP: Stereospecific reactivation of human brain and erythrocyte acetylcholinesterase inhibited by 1,2,2-trimethylpropyl methylphonofluoridate (soman). Biochim Biophys Acta 830:345–348, 1985

    Google Scholar 

  41. Kovach IM: Structure and dynamics of serine hydrolase-organophosphate adducts. J Enz Inhib 2:199–208, 1988

    Google Scholar 

  42. Ennis M, Shipley MT: Tonic activation of locus coeruleus neurons by systemic or intracoerulear microinjection of an irreversible acetylcholinesterase inhibitor: increased discharge rate and induction of C-fos. Exper Neur 118:164–177, 1992

    Google Scholar 

  43. Sidek HM, Nyquist-Battie C, Vanderkooi G: Inhibition of synaptosomd enzymes by local anesthetics. Biochim Biophys Acta 801:26–31, 1984

    Google Scholar 

  44. Spinedi A, Pacini L, Luly P: A study of the mechanism by which some amphiphilic drugs affect human erythrocyte acetylcholinesterase activity. Biochem J 261:569–573, 1989

    Google Scholar 

  45. Spinedi A, Pacini L, Limatola C, Luly P, Farias RN: Phenothiazines inhibit acetylcholinesterase by concentration-dependenttype kinetics. Biochem Pharmacol 44:1511–1514, 1992

    Google Scholar 

  46. Sussman JL, Harel M, Frolow F, Oefner C, Goldman A, Toker L, Silmam I: Atomic structure of acetylcholinesterase fromTorpedo californica: A prototypic acetylcholine-binding protein. Science 253:872–879, 1991

    Google Scholar 

  47. Dougherty DA, Stauffer DA: Acetylcholin binding by a synthetic receptor: Implications for biological recognition. Science 250: 1558–1560, 1990

    Google Scholar 

  48. Dennis M, Giraudat J, Kotzyba-Hibert F, Goeldner M, Hirth C, Chang J-Y, Lazure C, Chretien M, Cahngeux J-P: Amino acids of the Torpedo marmorata acetylcholine receptor a subunit labeled by a photoaffinity ligand for the acetylcholine binding site. Biochemistry 27:2346–2357, 1988

    Google Scholar 

  49. Galzi JL, Revah F, Black D, Goeldener M, Hirth C, Changeux J-P: Identification of a novel amino acid a-tyrosine 93 within the cholinergic ligands-binding sites of the acetylcholine receptor by photoaffinity labeling. J Biol Chem 265:10430–10437, 1990

    Google Scholar 

  50. Davie DR, Metzger H: Structure basis of antibody functure. Annu Rev Immunol 1:87–117, 1983

    Google Scholar 

  51. Adams LM, Foote SL: Effects of lacally infused pharmacological agents on spontaneous and sensory-evoked activity of locus coeruleus neurons. Brain Res Bull 21:395–400, 1988

    Google Scholar 

  52. Trevor AJ, Castagnoli N Jr, Singer TP: The formation of reactive intermediates in the MAO-catalyzed oxidation of the nigrostriatal toxin 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP). Toxicology 49:513–519, 1988

    Google Scholar 

  53. Albanese A, Butcher LL: Locus ceruleus somata contain both acetylcholinesterase and norepinephrine: direct histochemical demonstration on the same tissue section. Neurosci Lett 14:101–104

  54. Javitch JA, D'Amato RJ, Strittmatter SM, Snyder SH: Parkinsonism-inducing neurotoxin, N-methyl-4-phenyl-1,2,3,6-tetrahydropyridine: uptake of the metabolite N-methyl-4-phenylpyridine by dopamine neurons explains selective toxicity. Proc Natl Acad Sci USA 82:2173–2177, 1985

    Google Scholar 

  55. D'Amato RJ, Alexander GM, Schwartzman RJ, Kitt CA, Price DL, Sner SH: Neuromelanin: a role in MPTP-induced neurotoxicity. Life Sci 40:705–712, 1986

    Google Scholar 

  56. Zang LY, Misra HP: Superoxide radical production during the autoxidation of 1-methyl-4-phenyl-2,3-dihydropyridinium perchlorate. J Biol Chem 267:17547–17552, 1992

    Google Scholar 

  57. Zang LY, Misra HP: EPR kinetic studies of superoxide radicals generated during the autoxidation of 1-methyl-4-phenyl-2,3-dihydro-pyridinium, a bioactivated intermediate of Parkinsonian-inducing neurotoxin 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine. J Biol Chem 267:23601–23608, 1992

    Google Scholar 

  58. Finkelstein E, Rosen GM, Rauckman EJ: Production of hydroxyl radical by decomposition of superoxide spin adducts. Mol Pharmacol 21:262–265, 1982

    Google Scholar 

  59. Zang LY, Misra HP: Generation of reactive oxygen species during the monoamine oxidase-catalyzed oxidation of the neurotoxin, 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine. J Biol Chem, 1993, in press

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  1. Department of Biomedical Sciences, Virginia-Maryland Regional College of Veterinary Medicine, Virginia Polytechnic Institute and State University, 24061-0442, Blacksburg, VA, USA

    Lun-Yi Zang & Hara P. Misra

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  1. Lun-Yi Zang
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Zang, LY., Misra, H.P. Acetylcholinesterase inhibition by 1-methyl-4-phenylpyridinium ion, a bioactivated metabolite of MPTP. Mol Cell Biochem 126, 93–100 (1993). https://doi.org/10.1007/BF00925686

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