The role of membrane protein sulfhydryl groups in hydrogen peroxide-mediated membrane damage in human erythrocytes.
The role of membrane protein sulfhydryl groups in hydrogen peroxide-mediated membrane damage in human erythrocytes.
复制标题
膜蛋白巯基在过氧化氢介导的人红细胞膜损伤中的作用。
DOI：
10.1016/0005-2736(88)90245-3
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发表时间：
1988
期刊：
影响因子：
--
通讯作者：
Mohandas,N
中科院分区：
文献类型：
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作者：
Snyder,LM;Fortier,NL;Leb,L;McKenney,J;Trainor,J;Sheerin,H;Mohandas,N
用过氧化氢(H_2O_2)处理红细胞后，血影蛋白-血红蛋白复合体的形成与细胞形状的改变、膜变形性的降低以及单核细胞吞噬实验中抗IgM免疫球蛋白对修饰细胞的识别增加有关。预先用一氧化碳处理可完全抑制H_2O_2相关膜的变化，表明氧化血红蛋白在复合体的形成中起作用。此外，在无细胞体系中，N-乙基马来酰亚胺对纯化的血影蛋白的巯基(SH)的封闭显著减少了络合物的形成，这表明光影蛋白的SH基团在交联过程中起到了作用。本研究旨在探讨N-乙基马来酰亚胺阻断SH对氧化损伤的完整红细胞的作用。用浓度为0.1-0.2 mM的N-乙基马来酰亚胺处理红细胞，可降低脂质过氧化和血影蛋白的交联度。此外，N-乙基马来酰亚胺对细胞形态和膜变形性的影响较小，抗球蛋白血清对过氧化细胞的识别能力降低。N-乙基马来酰亚胺处理对高铁血红蛋白的形成没有影响。对14C标记的N-乙基马来酰亚胺的研究表明，超过50%的N-乙基马来酰亚胺被引入到血影蛋白中。用较高浓度的N-乙基马来酰亚胺(>0.2 mM)处理细胞后，膜功能障碍不依赖于H_2O_2。这些结果表明，阻断反应性SH基会降低血影蛋白与氧化珠蛋白的相互作用。这些数据和我们先前的观察表明，血影蛋白表面的SH基团在血红蛋白氧化诱导的血影蛋白-血红蛋白复合体的形成以及由此产生的对膜性能的有害影响中起着重要作用。
The formation of spectrin-hemoglobin complex following treatment of red cells with hydrogen peroxide (H 2 O 2) 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 H 2 O 2-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 14 C-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 H 2 O 2. 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.