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In vitro inhibition of human erythrocyte acetylcholinesterase (EC3.1.1.7) by an antineoplastic drug methotrexate

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Abstract

This work addresses the kinetic analysis of the interaction of methotrexate (MTX) with human erythrocyte membrane-bound acetylcholinesterase (AChE, EC 3.1. 1.7). It was found that the MTX effect was independent of time of incubation with AChE before the addition of substrate which proves its reversible action. The IC50 was determined, by three methods, to be 0.73 mM. The Michaelis-Menten constant (Ks) for the hydrolysis of acetylthiocholine iodide (ASCh) by AChE was 0.13 mM in the control system, a value decreased by 30–61% in the MTX treated systems. The Vmax was 1.27μtmole/min/mg protein for the control system while it was decreased by 44–77% in the MTX treated systems. The Linexveaver-Buck plot, Dixon plot, and their secondary replots indicated that the nature of the inhibition was of the linear mixed type, i.e. uncompetitive and noncompetitive. The values of Ki(slope) and KI(tntecept) were estimated as 1.67 and 0.34 mM, respectively.

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Abbreviations

AChE:

acetylcholinesterase

ASCh:

acetylthiocholine

Ks :

Michaelis-Menten constant

Vmax :

the limiting maximal velocity

Ki :

inhibition constant

MTX:

methotrexate

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Authors and Affiliations

  1. Department of Biochemistry, College of Science, King Saud University, P.O. Box 2455, 11451, Riyadh, Saudi Arabia

    Mohammad A. Kamal & Abdulaziz A. Al-Jafari

  2. Department of Pharmacy, Islamia University, Bahawalpur, Pakistan

    Faizul H. Nasim

Authors
  1. Mohammad A. Kamal
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  2. Faizul H. Nasim
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  3. Abdulaziz A. Al-Jafari
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Kamal, M.A., Nasim, F.H. & Al-Jafari, A.A. In vitro inhibition of human erythrocyte acetylcholinesterase (EC3.1.1.7) by an antineoplastic drug methotrexate. Mol Cell Biochem 159, 47–53 (1996). https://doi.org/10.1007/BF00226062

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  • DOI: https://doi.org/10.1007/BF00226062

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