Band 3 tyr-phosphorylation in normal and glucose-6-phospate dehydrogenase-deficient human erythrocytes.
Haemolysis is usually episodic in glucose-6-phosphate dehydrogenase (G6PD) deficiency, often triggered by a period of oxidative stress. In the present work, we investigate a possible biochemical mechanism underlying the enhanced susceptibility of G6PD deficient red blood cells (RBC) to oxidative stress. We analysed eight male subjects with Mediterranean glucose-6P-dehydrogenase deficiency (G6PDd), class II, for their ability in phosphorylating erythrocyte membrane band 3 following oxidative and osmotic stress. Our findings show that this sensitivity is connected to an early membrane band 3 Tyr-phosphorylation in the presence of diamide. However, since both Syk, and Lyn kinases, and SHP-2 phosphatase, mostly implicated in the band 3 P-Tyr level regulation, are alike in content and activity in normal and patient erythrocytes, an alteration in the membrane organization is likely the cause of the anomalous response to the oxidant. We report, in fact, that hypertonic-induced morphological change in G6PDd erythrocyte induces a higher membrane band 3 Tyr-phosphorylation, suggesting a pre-existing membrane alteration, likely due to the chronic lowering of the redox systems in patients. We also report that 1-chloro-2,4-dinitrobenzene-pre-treatment of normal red cells can alter the normal protein-protein and protein-membrane interaction under hypertonic rather than oxidative stress, thus partially resembling the response in patients, and that RBC may utilize a wider range of redox defence, under oxidative conditions, including, but not exclusively, NADPH and glutathione. On the whole, these results would encourage a different approach to the evaluation of the effects of pharmacological administration to patients, giving more attention to the possible drug-induced membrane alteration evidenced by the abnormal band 3 Tyr-phosphorylation.
Bordin
Luciana
L
Department of Biological Chemistry, University of Padova, Viale G. Colombo, 3, 35121, Padova, Italy.
Zen
Francesco
F
Ion-Popa
Florina
F
Barbetta
Micaela
M
Baggio
Bruno
B
Clari
Giulio
G
D000738:Androsterone / Q000494:pharmacology; D001457:Anion Exchange Protein 1, Erythrocyte / Q000378:metabolism*; D000975:Antioxidants / Q000378:metabolism; D003958:Diamide / Q000494:pharmacology; D004137:Dinitrochlorobenzene / Q000494:pharmacology; D004912:Erythrocytes / Q000187:drug effects / Q000201:enzymology*; D005955:Glucosephosphate Dehydrogenase Deficiency / Q000201:enzymology*; D005978:Glutathione / Q000378:metabolism; D006801:Humans; D008297:Male; D009249:NADP / Q000378:metabolism; D009997:Osmotic Pressure; D018384:Oxidative Stress; D010766:Phosphorylation; D017027:Protein Tyrosine Phosphatases / Q000378:metabolism; D014443:Tyrosine / Q000378:metabolism*
D001457:Anion Exchange Protein 1, Erythrocyte; D000975:Antioxidants; D004137:Dinitrochlorobenzene; D003958:Diamide; D014443:Tyrosine; D009249:NADP; D000738:Androsterone; D017027:Protein Tyrosine Phosphatases; D005978:Glutathione