Ward G M, Harrison L C
Diabetes. 1986 Jan;35(1):101-5. doi: 10.2337/diab.35.1.101.
The structure of the insulin receptor in intact human erythrocytes was defined using the techniques of disuccinimidyl suberate (DSS) cross-linking of 125I-insulin and surface [125I]iodination followed by receptor immunoprecipitation. In contrast to a recent report, we found the erythrocyte insulin receptor to be similar in structure to that in classic target tissues for insulin, consisting of at least three species of molecular weight approximately 295,000, 265,000, and 245,000, containing disulfide-linked subunits of molecular weight approximately 130,000 and 95,000. The interconversion of the three oligomeric forms could mediate changes in receptor affinity as postulated in other tissues. The 95,000 subunit was detected by immunoprecipitation only if surface iodination was performed in a Tris/Hepes buffer using lodogen and not if phosphate-buffered saline or lactoperoxidase iodination was used. These findings indicate that the lack of a bioeffect of insulin in erythrocytes is not explained by a gross defect in the structure of their insulin receptors. The apparent identity of the insulin receptor structure in erythrocytes and insulin target tissues provides a firmer basis for the use of erythrocytes in some circumstances to reflect insulin receptor status.
运用辛二酸二琥珀酰亚胺酯(DSS)交联125I胰岛素和表面[125I]碘化技术,随后进行受体免疫沉淀,确定了完整人红细胞中胰岛素受体的结构。与最近的一份报告相反,我们发现红细胞胰岛素受体的结构与胰岛素经典靶组织中的结构相似,由至少三种分子量约为295,000、265,000和245,000的分子组成,含有分子量约为130,000和95,000的二硫键连接亚基。正如在其他组织中所假设的那样,三种寡聚体形式的相互转化可能介导受体亲和力的变化。仅当在Tris/Hepes缓冲液中使用碘代甘氨酸进行表面碘化时,才能通过免疫沉淀检测到95,000亚基;如果使用磷酸盐缓冲盐水或乳过氧化物酶碘化,则无法检测到。这些发现表明,红细胞中胰岛素缺乏生物效应不能用其胰岛素受体结构的严重缺陷来解释。红细胞和胰岛素靶组织中胰岛素受体结构的明显一致性为在某些情况下使用红细胞来反映胰岛素受体状态提供了更坚实的基础。