Proteolysis in human erythrocytes is triggered only by selected oxidative stressing agents.
Human erythrocytes have been treated with different agents producing oxidative stress. Diamide, tetrathionate, chromate, cystamine and iodate preferentially influenced the cellular redox state by oxidation of free and protein thiol groups leaving the redox state of hemoglobin virtually unaffected. None of these compounds was able to stimulate the proteolysis. By contrast, phenylhydrazine, nitrite, hydrogen peroxide, ter-butylhydroperoxide, cumene hydroperoxide and copper-ascorbate caused a noticeable oxidation of hemoglobin to methemoglobin. These latter agents, except nitrite and copper-ascorbate, triggered proteolysis. Identical results have been obtained in a ghost-free hemolysate. The fraction containing the proteolytic activity was isolated from hemolysate and tested on native or oxidant-treated hemoglobin. The proteolysis was stimulated by all agents able to produce methemoglobin. It is concluded that proteolysis correlated to an unbalance of cellular redox state. The results obtained with isolated and recombined fractions suggests that increased proteolysis does not depend on the removal of the effect of protease(s) inhibitor(s). Since all agents stimulating proteolysis are able to generate free radicals, it seems that protein breakdown is triggered by the direct effect of these intermediates on proteins (mostly hemoglobin) without the involvement of radical species produced in the membranes by action of organic hydroperoxides. In addition, since nitrite and copper-ascorbate, which also oxidize hemoglobin by radical generation, are unable to stimulate proteolysis, it should be concluded that protein degradation induced by oxidative stress is a complex event induced only by specific agents.
D004912:Erythrocytes / Q000187:drug effects / Q000378:metabolism*; D005609:Free Radicals; D006454:Hemoglobins / Q000378:metabolism; D006801:Humans; D006861:Hydrogen Peroxide / Q000494:pharmacology; D066298:In Vitro Techniques; D010084:Oxidation-Reduction; D010447:Peptide Hydrolases / Q000097:blood*; D010659:Phenylhydrazines / Q000494:pharmacology; D011480:Protease Inhibitors / Q000097:blood; D013438:Sulfhydryl Compounds / Q000097:blood
D005609:Free Radicals; D006454:Hemoglobins; D010659:Phenylhydrazines; D011480:Protease Inhibitors; D013438:Sulfhydryl Compounds; C030299:phenylhydrazine; D006861:Hydrogen Peroxide; D010447:Peptide Hydrolases