Interactions between peroxiredoxin 2, hemichrome and the erythrocyte membrane.
Peroxiredoxin 2 (Prx2) is an abundant antioxidant protein in erythrocytes that protects against hemolytic anemia resulting from hemoglobin oxidation and Heinz body formation. A small fraction of Prx2 is bound to the cell membrane, but the mechanism and relevance of binding are not clear. We have investigated Prx2 interactions with the erythrocyte membrane and oxidized hemoglobin and whether these interactions are dependent on Prx2 redox state. Membrane binding of Prx2 in erythrocytes decreased when the cells were treated with H2O2, but studies with purified Prx2 and isolated ghosts showed that the interaction was independent of Prx2 redox state. Hemoglobin oxidation leads to the formation of hemichrome, a denatured form of the protein that binds to Band3 protein in the cell membrane as part of the senescence process and is a precursor of Heinz bodies. Hemichrome competed with Prx2 and decreased Prx2 binding to the membrane, potentially explaining the decreased binding in oxidant-exposed cells. The increased membrane binding of Prx2 seen with increasing intracellular calcium was less sensitive to H2O2or hemichrome, suggesting an alternative mode of binding. Prx2 was also shown to exhibit chaperone-like activity by retarding the precipitation of pre-formed hemichrome. Our results suggest that Prx2, by restricting membrane binding of hemichrome, could impede Band3 clustering and exposure of senescence antigens. This mechanism, plus the observed chaperone activity for oxidized hemoglobin, may help protect against hemolytic anemia.
a Department of Pathology , Centre for Free Radical Research, University of Otago , Christchurch , New Zealand.
Simone B
SB
Bayer
a Department of Pathology , Centre for Free Radical Research, University of Otago , Christchurch , New Zealand.
Felicia M
FM
Low
a Department of Pathology , Centre for Free Radical Research, University of Otago , Christchurch , New Zealand.
Mark B
MB
Hampton
a Department of Pathology , Centre for Free Radical Research, University of Otago , Christchurch , New Zealand.
Christine C
CC
Winterbourn
D000975:Antioxidants; D004910:Erythrocyte Membrane / Q000187:drug effects*; D006420:Hemeproteins / Q000378:metabolism*; D006454:Hemoglobins / Q000378:metabolism; D054464:Peroxiredoxins / Q000378:metabolism*
D000975:Antioxidants; D006420:Hemeproteins; D006454:Hemoglobins; C001332:hemichrome; D054464:Peroxiredoxins
Antioxidant defense; Heinz body; chaperone; hemoglobin oxidation; phenylhydrazine