13C and 31P NMR studies of glucose and 2-deoxyglucose metabolism in normal and enzyme-deficient human erythrocytes.
The flux of 13C-labeled glucose through the Embden-Meyerhof and pentose phosphate pathways was studied by 13C NMR in intact erythrocytes isolated from normal subjects or from patients suffering of glucose-6-phosphate dehydrogenase (G6PD, EC 1.1.1.49) deficiency. Similar rates of glucose catabolism and similar fluxes of the 13C-label into 2,3-bisphosphoglycerate and lactate were found, under basal conditions, in normal and in G6PD-deficient erythrocytes incubated in the presence of either [1-13C]- or D[6-13C]glucose. Exposure to oxidative stress by preincubation with tert-butylhydroperoxide induced in normal, but not in G6PD-deficient erythrocytes, a significant enhancement of glucose consumption, as well as a substantial reduction in 13C-label transfer from C1-glucose into lactate. It was also possible, by 31P NMR, to evaluate the conversion of 2-deoxyglucose to its phosphate-containing metabolites. The oxidation and subsequent decarboxylation of 2-deoxyglucose-6-phosphate was assessed in reconstituted systems and could subsequently be evidenced also in ethanolic extracts from normal (but not from G6PD-deficient) erythrocytes which had been exposed to oxidative stress. The results indicate that, in terms of glucose flux through the glycolytic pathway, there is little or no difference between normal and G6PD-deficient erythrocytes, regardless of previous exposure to oxidative stress. Faster consumption of either glucose or 2-deoxyglucose is induced, only in normal cells, by treatment with tert-butylhydroperoxide, essentially as a consequence of the activation of the pentose-phosphate pathway.
Ferretti
A
A
Laboratorio di Biologia Cellulare, Istituto Superiore di Sanità, Rome, Italy.
Bozzi
A
A
Di Vito
M
M
Podo
F
F
Strom
R
R
D019794:2,3-Diphosphoglycerate; D001786:Blood Glucose / Q000378:metabolism*; D002247:Carbon Isotopes; D002851:Chromatography, High Pressure Liquid; D003847:Deoxyglucose / Q000378:metabolism*; D004163:Diphosphoglyceric Acids / Q000097:blood; D004912:Erythrocytes / Q000187:drug effects / Q000201:enzymology*; D005955:Glucosephosphate Dehydrogenase Deficiency / Q000097:blood*; D006801:Humans; D009682:Magnetic Resonance Spectroscopy; D010545:Peroxides / Q000494:pharmacology; D010758:Phosphorus; D020122:tert-Butylhydroperoxide
D001786:Blood Glucose; D002247:Carbon Isotopes; D004163:Diphosphoglyceric Acids; D010545:Peroxides; D019794:2,3-Diphosphoglycerate; D010758:Phosphorus; D020122:tert-Butylhydroperoxide; D003847:Deoxyglucose