Original Article

European Journal of Clinical Nutrition (2006) 60, 593–597. doi:10.1038/sj.ejcn.1602355; published online 14 December 2005

The effect of L-cysteine and glutathione on inhibition of Na+, K+-ATPase activity by aspartame metabolites in human erythrocyte membrane

Guarantors: KH Schulpis and S Tsakiris.

Contributors: KHS, ST, IP, TP and ST.

K H Schulpis1, I Papassotiriou2, T Parthimos3, T Tsakiris3 and S Tsakiris3

  1. 1Institute of Child Health, Research Center, 'Aghia Sophia' Children's Hospital, Athens, Greece
  2. 2Department of Clinical Biochemistry, 'Aghia Sophia' Children's Hospital, Athens, Greece
  3. 3Department of Experimental Physiology, Medical School, University of Athens, Athens, Greece

Correspondence: Professor S Tsakiris, Department of Experimental Physiology, Medical School, University of Athens, PO Box 65257, GR 15401 Athens, Greece. E-mail: stsakir@cc.uoa.gr

Received 27 June 2005; Revised 31 August 2005; Accepted 14 September 2005; Published online 14 December 2005.

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Abstract

Background:

 

Reports have implicated Aspartame (N-L-a-aspartyl-L-phenylalanine methyl ester, ASP) in neurological problems.

Aim:

 

To evaluate Na+, K+-ATPase activities in human erythrocyte membranes after incubation with the ASP metabolites, phenylalanine (Phe), methanol (MeOH) and aspartic acid (Asp).

Methods:

 

Erythrocyte membranes were obtained from 12 healthy individuals and were incubated at 37°C for 1 h with the sum or each of the ASP metabolites separately, which are commonly measured in blood after ASP ingestion. Na+, K+-ATPase and Mg2+-ATPase activities were measured spectrophotometrically.

Results:

 

Membrane Mg2+-ATPase activity was not altered. The sum of ASP metabolite concentrations corresponding to 34, 150 or 200 mg/kg of the sweetener ingestion resulted in an inhibition of the membrane Na+, K+-ATPase by -30, -40, -48%, respectively. MeOH concentrations of 0.14, 0.60 or 0.80 mM decreased the enzyme activity by -25, -38, -43%, respectively. Asp concentrations of 2.80, 7.60 or 10.0 mM inhibited membrane Na+, K+-ATPase by -26, -40, -46%, respectively. Phe concentrations of 0.14, 0.35 or 0.50 mM reduced the enzyme activity by -24, -44, -48%, respectively. Preincubation with L-cysteine or reduced glutathione (GSH) completely or partially restored the inhibited membrane Na+, K+-ATPase activity by high or toxic ASP metabolite concentrations.

Conclusions:

 

Low concentrations of ASP metabolites had no effect on Na+, K+-ATPase activity. High or abuse concentrations of ASP hydrolysis products significantly decreased the membrane enzyme activity, which was completely or partially prevented by L-cysteine or reduced GSH.

Keywords:

Aspartame, methanol, phenylalanine, aspartic acid, Na+, K+-ATPase, antioxidants

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