Lysko K A, Carlson R, Taverna R, Snow J, Brandts J F
Biochemistry. 1981 Sep 15;20(19):5570-6. doi: 10.1021/bi00522a034.
In this study, it is shown that systematic temperature-induced protein aggregation occurs on the erythrocyte membrane by intermolecular disulfide bond formation. Specific protein bands disappear from acrylamide gel profiles over rather narrow temperature regions. The aggregation appears to be the result of irreversible structural transitions of the membrane, which can be seen in a sensitive scanning calorimeter. When this method of thermal gel analysis is used, the results suggest that spectrin is a participant in the A transition, that bands 2.1, 4.1, and 4.2 and the cytoplasma portion of 3 are involved in the B transition, and that the transmembrane portion of band 3 may undergo changes in the C transition, previously shown to occur in the anion transport domain of the membrane. The aggregation of specific proteins in the narrow temperature region of these transitions persists as the transitions are moved around on the temperature axis by varying solution conditions. The assignment of particular proteins to specific transitions is reinforced by selective extraction of membrane proteins. Large variations in both the calorimetry and the aggregation pattern occur as salt concentration is increased from 77 mosm to 310 mosm, which is manifested in the splitting of the B transition into two separate transitions, B1 and B2. It is speculated that this occurs as the result of a structural change which may involve components of the cytoskeletal network.
本研究表明，通过分子间二硫键的形成，系统性温度诱导的蛋白质聚集发生在红细胞膜上。在相当窄的温度区域内，特定的蛋白质条带从丙烯酰胺凝胶图谱中消失。这种聚集似乎是膜不可逆结构转变的结果，这在灵敏的扫描量热仪中可以看到。当使用这种热凝胶分析方法时，结果表明血影蛋白参与A转变，2.1、4.1和4.2条带以及3条带的细胞质部分参与B转变，并且3条带的跨膜部分可能在C转变中发生变化，先前已证明这种变化发生在膜的阴离子转运结构域。随着通过改变溶液条件使转变在温度轴上移动，这些转变的窄温度区域内特定蛋白质的聚集持续存在。通过对膜蛋白的选择性提取，特定蛋白质与特定转变的对应关系得到了加强。当盐浓度从77毫渗摩尔增加到310毫渗摩尔时，量热法和聚集模式都出现了很大变化，这表现为B转变分裂为两个独立的转变，即B1和B2。据推测，这是由于可能涉及细胞骨架网络成分的结构变化导致的。
