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The formation of spectrin-hemoglobin complex following treatment of red cells with hydrogen peroxide (H2O2) has previously been shown to be associated with alterations in cell shape, decreased membrane deformability and increased recognition of modified cells by anti-IgM immunoglobulin in a phagocytic assay by monocytes. Prior treatment with carbon monoxide completely inhibited the H2O2-associated membrane changes, indicating a role for oxidized hemoglobin in the complex formation. Also, in a cell-free system, blockage of sulfhydryl (SH) groups on purified spectrin by N-ethylmaleimide significantly reduced the complex formation, suggesting a role for SH groups of spectrin in crosslinking process. The present study was undertaken to examine the role of SH blockade by N-ethylmaleimide on intact red cells undergoing oxidative damage. Pretreatment of erythrocytes with N-ethylmaleimide at concentrations ranging from 0.1 to 0.2 mM resulted in decreased lipid peroxidation and spectrin hemoglobin crosslinking. Moreover, pretreatment with N-ethylmaleimide resulted in less marked alterations in cell shape and membrane deformability as well as reduced recognition of peroxidized cells by antiglobulin serum. N-Ethylmaleimide treatment had no effect on methemoglobin formation. Studies with 14C-labeled N-ethylmaleimide showed that over 50% of N-ethylmaleimide was incorporated into spectrin. Pretreatment of cells with higher concentrations of N-ethylmaleimide (over 0.2 mM) was associated with membrane dysfunction independent of H2O2. These results imply that blocking of reactive SH groups leads to reduced interaction of spectrin with oxidized globin. These data, along with our prior observations, indicate that SH groups on spectrin play an important role in hemoglobin oxidation-induced formation of spectrin-hemoglobin complex and the resultant deleterious effects on membrane properties.
L M Snyder, and
N L Fortier, and
L Leb, and
J McKenney, and
J Trainor, and
H Sheerin, and
N Mohandas
July 1967,
Biochimica et biophysica acta,
L M Snyder, and
N L Fortier, and
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J McKenney, and
J Trainor, and
H Sheerin, and
N Mohandas
August 1985,
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L M Snyder, and
N L Fortier, and
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H Sheerin, and
N Mohandas
February 1996,
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L M Snyder, and
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J McKenney, and
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N Mohandas
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L M Snyder, and
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July 2003,
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L M Snyder, and
N L Fortier, and
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N Mohandas
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L M Snyder, and
N L Fortier, and
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J Trainor, and
H Sheerin, and
N Mohandas
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L M Snyder, and
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May 1991,
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L M Snyder, and
N L Fortier, and
L Leb, and
J McKenney, and
J Trainor, and
H Sheerin, and
N Mohandas
January 1968,
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L M Snyder, and
N L Fortier, and
L Leb, and
J McKenney, and
J Trainor, and
H Sheerin, and
N Mohandas
July 1971,
Biochemical and biophysical research communications,
