Oron Y, Galasko G, Larner J
Mol Cell Biochem. 1980 Nov 20;32(3):161-7. doi: 10.1007/BF00227443.
Incubation of intact mouse diaphragms with insulin in the absence of glucose resulted in a rapid inhibition of the subsequent cell-free phosphorylation of endogenous protein substrates in tissue extracts. The phosphorylation of added histone was inhibited to a lesser extent. The inhibition was observed both in the absence and in the presence of added cyclic 3'5' adenosine monophosphate. Acrylamide gel electrophoresis of phosphorylation products revealed a number of major phosphorylated polypeptides. The phosphorylation of several polypeptides was inhibited following short treatment with insulin. These results represent a novel experimental approach to the elucidation of the mechanism of the action of insulin and are consistent with our hypothesis that the inhibition of protein kinase activities in the tissue may be the first step in this mechanism.
在无葡萄糖存在的情况下，将完整的小鼠膈肌与胰岛素一起温育，会导致随后组织提取物中内源性蛋白质底物的无细胞磷酸化迅速受到抑制。添加的组蛋白的磷酸化受到的抑制程度较小。在添加或不添加环磷腺苷的情况下均观察到这种抑制作用。磷酸化产物的丙烯酰胺凝胶电泳显示出一些主要的磷酸化多肽。用胰岛素短时间处理后，几种多肽的磷酸化受到抑制。这些结果代表了一种阐明胰岛素作用机制的新实验方法，并且与我们的假设一致，即组织中蛋白激酶活性的抑制可能是该机制的第一步。
