Characterization of skeletal muscle insulin receptor.
Characterization of skeletal muscle insulin receptor.
复制标题
骨骼肌胰岛素受体的表征。
DOI：
10.1016/0003-9861(85)90611-3
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发表时间：
1985
影响因子：
3.9
通讯作者：
Larner,J
中科院分区：
文献类型：
--
作者：
Dubler,RE;Whipple,JH;Larner,J
对大鼠骨骼肌的胰岛素受体进行了表征。用光活性胰岛素类似物 125 I[N-εB29-单叠氮基苯甲酰基]-胰岛素处理肌肉膜，显示出 135,000 Da 的单个蛋白带，即 α 亚基。对总膜蛋白进行碘化，然后进行 Triton X-100 溶解和免疫沉淀，证明存在 90,000 Da 的蛋白带（β 亚基）以及 190,000 Da 的蛋白带（可能是受体前体）。在部分纯化的受体制剂中，β亚基表现出剂量依赖性、胰岛素刺激的磷酸化，仅将磷酸盐掺入酪氨酸残基中，这也在 190,000-Da 受体前体中观察到。纯化的质膜含有大量的胰岛素降解活性，在进行胰岛素结合研究之前必须将其灭活。如果不阻止胰岛素的降解，则观察到在未标记的胰岛素存在下结合明显增强。
The insulin receptor from rat skeletal muscle was characterized. Treatment of muscle membranes with the photoactive insulin analog,125I[N-ϵB29-monoazidobenzoyl]-insulin revealed a single protein band of 135,000 Da, the α subunit. Iodination of total membrane protein followed by Triton X-100 solubilization and immunoprecipitation demonstrated the presence of a protein band of 90,000 Da, the β subunit, together with a protein band of 190,000 Da, which may be the receptor precursor. In partially purified receptor preparations, the β subunit exhibited dose-dependent, insulinstimulated phosphorylation with incorporation of phosphate solely into tyrosine residues, which was also observed in the 190,000-Da receptor precursor. Purified plasma membranes contained a large amount of insulin-degrading activity which had to be inactivated prior to performing insulin-binding studies. If degradation of insulin was not prevented, apparent enhanced binding in the presence of unlabeled insulin was observed.