Reduced acetylcholinesterase activity in erythrocyte membranes from patients with phenylketonuria

Abstract

Objective: a) To evaluate acetylcholinesterase (AChE) activities in erythrocyte membranes from phenylketonuric (PKU) patients and controls and to correlate with their plasma phenylalanine (Phe), tyrosine (Tyr), alanine (Ala) and dopamine (DA) levels. b) To determine the in vitro effects of Phe, Ala and Phe plus Ala on their AChE activities. Design and Methods: AChE activities were determined spectrophotometrically in erythrocyte membranes from PKU children ( n = 12) adhering to their diet (group A), from 11 “off diet” (group B) and from 23 controls. Their plasma amino acids were evaluated with an amino acid analyser and DA with an HPLC method. Ala (1.8 mM) and/or Phe (1.8 mM) were added in the enzyme incubation medium from controls, whereas only Ala was added in that from group B. Results: AChE activity (1.19 ± 0.05 Δ OD/min × mg protein), Tyr (46 ± 17 μmol/L) and DA (56 ± 18 μmol/L) were remarkably decreased by about 60% in group B as compared to those of group A ( 3.01 ± 0.18 Δ OD/min × mg protein, 115 ± 39 μmol/L, 137 ± 29 μmol/L, respectively, p < 0.001) and controls ( 3.13 ± 0.16 Δ OD/min × mg protein, 117 ± 44 μmol/L, 142 ± 22 μmol/L, respectively, p < 0.001). Phe negatively correlated with AChE activity and positively with plasma Tyr and DA. Ala reversed the inhibited AChE by Phe in erythrocyte membranes from healthy children to control values, whereas no reverse effect was observed on the enzyme activity from PKU patients. Conclusions: a) The low levels of DA and its precursor Tyr are due to the high Phe blood levels, as a consequence of the decreased activity of Phe-hydroxylase in the liver of our patients. So, high Phe blood levels inhibit AChE in PKU patients, probably resulting in higher acetylcholine concentrations. b) Determination of AChE in erythrocyte membranes from PKU could be a useful marker for the neurotoxic effects of Phe.