Linkage of cytosolic peroxiredoxin 2 to erythrocyte membrane imposed by hydrogen peroxide-induced oxidative stress.
Human erythrocyte peroxiredoxin 2 (Prx2) is a typical 2-cys cytosolic peroxiredoxin with thiol-dependent hydrogen peroxide scavenger activity. In a previous work, we reported Prx2 erythrocyte membrane linkage in some Hereditary Spherocytosis patients and that it seemed to be related to oxidative stress. The aim of the present work was to determine if Prx2 linkage to erythrocyte membrane could be induced by oxidative stress mediated by H(2)O(2) and to further understand how and why this process occurs. We performed in vitro assays in which catalase or both Hb autoxidation and catalase were inhibited, under H(2)O(2)-induced oxidative stress conditions. Erythrocyte membrane linked Prx2 was detected by immunoblotting and quantified by densitometry. As oxidative stress markers, we determined membrane bound hemoglobin and lipid peroxidation, and we found that their values increased with H(2)O(2) concentration. Prx2 linkage to the membrane also rose with increasing H(2)O(2) concentration, and was only observed when the oxidized form of the enzyme was present in the cytosol. Oxidized Hb and Prx2 membrane linkages appear to be independent processes, although, both result from oxidative stress and may be useful as oxidative stress and/or erythrocyte damage/senescence markers.
Faculdade de Farmácia, Universidade do Porto, Porto, Portugal.
Susana
S
Rocha
Elísio
E
Costa
Susana
S
Coimbra
Henrique
H
Nascimento
Cristina
C
Catarino
Petronila
P
Rocha-Pereira
Alexandre
A
Quintanilha
Luís
L
Belo
Alice
A
Santos-Silva
D002374:Catalase / Q000037:antagonists & inhibitors; D004910:Erythrocyte Membrane / Q000737:chemistry* / Q000187:drug effects*; D004912:Erythrocytes / Q000187:drug effects / Q000378:metabolism; D006454:Hemoglobins / Q000032:analysis / Q000378:metabolism; D006801:Humans; D006861:Hydrogen Peroxide / Q000494:pharmacology*; D015151:Immunoblotting; D015227:Lipid Peroxidation / Q000187:drug effects; D010084:Oxidation-Reduction; D018384:Oxidative Stress; D054464:Peroxiredoxins / Q000032:analysis* / Q000378:metabolism*
D006454:Hemoglobins; D006861:Hydrogen Peroxide; D054464:Peroxiredoxins; D002374:Catalase