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. 2008 Jul;12(3):162-7.

Abnormal glyceraldehyde-3-phosphate dehydrogenase binding and glycolytic flux in Autosomal Dominant Polycystic Kidney Disease after a mild oxidative stress

C Dioudis  1 , G Dimitrios, T H Thomas, I C West
Affiliations

Abnormal glyceraldehyde-3-phosphate dehydrogenase binding and glycolytic flux in Autosomal Dominant Polycystic Kidney Disease after a mild oxidative stress

C Dioudis et al. Hippokratia. 2008 Jul.

Abstract

Aim: The aim of this study was, a) to investigate the effect of mild oxidative stress on glycolytic flux and glyceraldehyde-3-phosphate dehydrogenase (GAPDH) binding in erythrocytes from patients with autosomal dominant polycystic kidney disease (ADPKD), and b) to examine whether the modulation of GAPDH-binding to the red cell membrane leads to changes in glycolytic flux.

Patients and methods: The rate of lactate production in intact erythrocytes and the GAPDH/actin ratio in erythrocyte ghost membranes were measured before and after treating cells with t-butyl hydroperoxide or N-ethylmaleimide (NEM) in 13 ADPKD patients and 12 controls.

Results: t-bytyl hydro-peroxide had a significant effect on both lactate production and GAPDH/actin ratio in healthy subjects, but it had essentially no effect on ADPKD patients in which both parameters already resembled those of the peroxide-treated controls. NEM treatment after 300 sec had a very significant effect on both lactate production and GAPDH/actin ratio in both patient and control cells. However, after 10 sec the effect on GAPDH/actin ratio was only significant in the erythrocytes of ADPKD patients. In every experiment glycolytic lactate production correlated negatively with membrane-bound GAPDH/actin ratio.

Conclusions: We conclude that glycolytic flux and GAPDH binding in erythrocytes from ADPKD patients respond abnormally to both a mild oxidative stress and brief exposure to NEM.

Keywords: autosomal dominant polycystic kidney disease; glyceraldehyde-3-phosphate dehydrogenase; glycolysis; oxidative stress.

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Figures

Figure 1.
Figure 1.. The effect of different concentrations of t-butyl hydroperoxide on rate of glycolytic lactate production by intact erythrocytes from healthy subjects.
Figure 2.
Figure 2.. Time-course of the effect of 1 mM N-ethylmaleimide (NEM), at pH 6 and 0o C, on rate of glycolytic lactate production in intact healthy erythrocytes.
Figure 3.
Figure 3.. Figure 3: The effect of different concentrations of N-ethylmaleimide on lactate production rate in intact healthy erythrocytes.
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