Peroxidative membrane damage in human erythrocytes induced by a concerted action of iodoacetate, vanadate and ferricyanide.
Human erythrocytes incubated without substrate in the presence of iodoacetate (0.2 mM), vanadate (0.5 mM) and ferricyanide (5 mM) form aqueous membrane leaks of equivalent radii of 0.5-0.8 nm leading to complete colloid-osmotic lysis within 180 min. All three components are indispensable for the effect. Inosine but not glucose markedly enhances the rate of hemolysis. These effects are due to oxidative damage, as indicated by concomitant destruction of polyunsaturated fatty acids and suppression of both effects by radical scavengers. Hemoglobin is not oxidized under these conditions. GSH and membrane SH levels remain almost normal, and no crosslinking or irreversible aggregation of membrane proteins is observed. In the absence of O2 no membrane damage can be observed. It is proposed that radical formation originates from reduction of O2 by NADPH, analogous to processes described in microsomal membranes. NADH seems not to be involved, since leak formation occurs in spite of the blockage of NADH formation by iodoacetate. Vanadate and ferricyanide are probably required to amplify the peroxidative reaction sufficiently to overcome the cellular antioxidative capacity.
D004910:Erythrocyte Membrane / Q000187:drug effects* / Q000378:metabolism; D005231:Fatty Acids, Unsaturated / Q000097:blood; D005292:Ferricyanides / Q000494:pharmacology*; D005609:Free Radicals; D006461:Hemolysis / Q000187:drug effects; D006801:Humans; D007288:Inosine / Q000494:pharmacology; D007461:Iodoacetates / Q000494:pharmacology*; D019807:Iodoacetic Acid; D008051:Lipid Bilayers / Q000097:blood; D008563:Membrane Lipids / Q000097:blood; D010084:Oxidation-Reduction; D010100:Oxygen / Q000097:blood; D010545:Peroxides / Q000097:blood*; D014639:Vanadium / Q000494:pharmacology*
D005231:Fatty Acids, Unsaturated; D005292:Ferricyanides; D005609:Free Radicals; D007461:Iodoacetates; D008051:Lipid Bilayers; D008563:Membrane Lipids; D010545:Peroxides; D014639:Vanadium; D007288:Inosine; D010100:Oxygen; C028033:potassium ferricyanide; D019807:Iodoacetic Acid