Sullivan S G, Stern A
Biochim Biophys Acta. 1984 Jul 25;774(2):215-20. doi: 10.1016/0005-2736(84)90294-3.
Red cells were incubated in the presence of t-butyl hydroperoxide and effects on red cell membrane proteins were studied by SDS-polyacrylamide gel electrophoresis. t-Butyl hydroperoxide caused diminution in intensity of all major cytoskeletal bands with the concomitant formation of high molecular weight material. Membrane glycoproteins were unaffected. t-Butyl hydroperoxide increased hemoglobin binding to ghosts. After dissolution in SDS and beta-mercaptoethanol, membrane-bound hemoglobin appeared on the gels in the form of monomers and crosslinked polymers of hemoglobin or globin chains. Crosslinking was partially prevented by metabolism of t-butyl hydroperoxide by the hexose monophosphate shunt except in methemoglobin-containing red cells where reaction with methemoglobin accounted for most of the consumption of t-butyl hydroperoxide. Metal chelators, deferoxamine mesylate and diethylenetriaminepentaacetic acid, had no effect on membrane protein changes. Butylated hydroxytoluene, diphenylamine and ascorbate, compounds that inhibit t-butyl hydroperoxide-induced red cell membrane lipid peroxidation, had no effect on t-butyl hydroperoxide-induced membrane protein changes. These results suggest that membrane proteins and membrane lipids have different mechanisms of peroxidant damage.
将红细胞置于叔丁基过氧化氢存在的环境中孵育，并通过十二烷基硫酸钠-聚丙烯酰胺凝胶电泳研究其对红细胞膜蛋白的影响。叔丁基过氧化氢导致所有主要细胞骨架条带的强度减弱，同时形成高分子量物质。膜糖蛋白未受影响。叔丁基过氧化氢增加了血红蛋白与血影的结合。在十二烷基硫酸钠和β-巯基乙醇中溶解后，膜结合血红蛋白以血红蛋白或珠蛋白链的单体和交联聚合物形式出现在凝胶上。除了含高铁血红蛋白的红细胞中与高铁血红蛋白的反应占叔丁基过氧化氢消耗的大部分外，磷酸己糖旁路对叔丁基过氧化氢的代谢部分阻止了交联。金属螯合剂甲磺酸去铁胺和二乙烯三胺五乙酸对膜蛋白变化没有影响。丁基化羟基甲苯、二苯胺和抗坏血酸盐，这些抑制叔丁基过氧化氢诱导的红细胞膜脂质过氧化的化合物，对叔丁基过氧化氢诱导的膜蛋白变化没有影响。这些结果表明，膜蛋白和膜脂质具有不同的过氧化物损伤机制。
