Green F A, Jung C Y, Cuppoletti J, Owens N
Biochim Biophys Acta. 1981 Nov 6;648(2):225-30. doi: 10.1016/0005-2736(81)90038-9.
Hypertonic cryohemolysis is defined as the lysis of erythrocytes in a hypertonic environment when the temperature is lowered from above 15-18 degrees C below that temperature. This has been found to be a general phenomenon (that is, whether the solute is charged or not), to exhibit interesting temperature characteristics and to be preventable by agents such as valinomycin which tend to dissipate the concentration gradient across the cell membrane. As yet, no clear information is available to translate this phenomenon to the molecular level and to relate it to current structure/function concepts in the erythrocyte membrane. In this study, data are presented which would indicate on the basis of two entirely separate methodologies that the spectrin-actin cytoskeletal framework is involved in this phenomenon. The first of these methodologies is based on radiation-induced ablation of cryohemolysis and indicates that an intact macromolecular complex of an order of 16000 000 daltons is required for cryohemolysis with hypertonic NaCl. The second methodology is based on selective cross-linking of spectrin and actin in the agent diamide, which resulted in concentration-dependent suppression of cryohemolysis. Polyacrylamide gel electrophoresis of the erythrocyte from diamide-treated cells showed intense protein aggregation with loss of spectrin-actin and bands 4.1, 4.2. We conclude that the spectrin-actin cytoskeletal system possibly including its interaction with phospholipids is the key to the phenomenon of hypertonic cryohemolysis.
高渗性冷冻溶血被定义为当温度从15 - 18摄氏度以上降至该温度以下时，红细胞在高渗环境中的溶解。已发现这是一种普遍现象（即无论溶质是否带电），呈现出有趣的温度特性，并且可被诸如缬氨霉素之类倾向于消除跨细胞膜浓度梯度的试剂所阻止。到目前为止，尚无明确信息可将此现象转化到分子水平并将其与红细胞膜当前的结构/功能概念联系起来。在本研究中，基于两种完全不同的方法呈现的数据表明，血影蛋白 - 肌动蛋白细胞骨架框架参与了这一现象。其中第一种方法基于辐射诱导的冷冻溶血消融，表明对于用高渗氯化钠进行的冷冻溶血，需要一个16000000道尔顿量级的完整大分子复合物。第二种方法基于在二酰胺试剂中血影蛋白和肌动蛋白的选择性交联，这导致了冷冻溶血的浓度依赖性抑制。对经二酰胺处理的细胞的红细胞进行聚丙烯酰胺凝胶电泳显示，蛋白质强烈聚集，血影蛋白 - 肌动蛋白以及4.1、4.2条带消失。我们得出结论，血影蛋白 - 肌动蛋白细胞骨架系统可能包括其与磷脂的相互作用是高渗性冷冻溶血现象的关键。
