The role of oxidized nicotinamide adenine dinucleotide in fluoride inhibition of active sodium transport in human erythrocytes.
The rate coefficient for (22)Na release from previously labeled human erythrocytes was determined in the presence of 0.1-10 mM sodium fluoride (F). The oxidized nicotinamide adenine dinucleotide (NAD(+)) level at the end of 2 hr of incubation in tris(hydroxymethyl)aminomethane (Tris)-Ringer medium was also measured. Both parameters decreased proportionately as F concentration was raised. Both F-induced changes were immediate and were reversed by 10 mM pyruvate. The decrease in NAD(+) concentration following enolase inhibition by F is attributed to a diminished rate of formation in the reaction catalyzed by lactic dehydrogenase (LDH) with undiminished continued utilization in the reaction catalyzed by glyceraldehyde-3-phosphate dehydrogenase (GAPDH). It is postulated that the NAD(+) lowering limited the GAPDH step, resulting in proportionate decreases in the rates of phosphoglycerate kinase (PGK) and Na,K-dependent adenosine triphosphatase (Na,K-ATPase), a reaction sequence thought to link glycolysis with active Na extrusion. Adding pyruvate with F increased NAD(+) production at the LDH step, thus reactivating GAPDH, PGK, and Na,K-ATPase and leading to the observed restoration of (22)Na release. The results suggest, therefore, that F inhibits active Na transport in intact human erythrocytes indirectly through a lowering of NAD(+), although, direct inhibition of the Na,K-ATPase by F may possibly occur simultaneously.
D000251:Adenosine Triphosphatases / Q000037:antagonists & inhibitors; D000328:Adult; D001693:Biological Transport, Active; D004912:Erythrocytes / Q000187:drug effects / Q000201:enzymology / Q000378:metabolism*; D005459:Fluorides / Q000494:pharmacology*; D006019:Glycolysis; D006801:Humans; D008297:Male; D009243:NAD / Q000032:analysis*; D010042:Ouabain / Q000494:pharmacology; D010751:Phosphopyruvate Hydratase / Q000037:antagonists & inhibitors; D011773:Pyruvates / Q000494:pharmacology; D012964:Sodium / Q000378:metabolism*; D012975:Sodium Isotopes; D013997:Time Factors
D011773:Pyruvates; D012975:Sodium Isotopes; D009243:NAD; D010042:Ouabain; D012964:Sodium; D000251:Adenosine Triphosphatases; D010751:Phosphopyruvate Hydratase; D005459:Fluorides