GLUTATHIONE AND THE REACTIVITY OF INSULIN RECEPTOR SULFHYDRYLS

本项目的目的有两个：(1)评估监管 位于胰岛素受体胞浆区的活性硫醇(S) 通过改变谷胱甘肽氧化还原循环的试剂；和(2)执行 硫醇-生物素标记的胰岛素受体裂解测定 活性硫醇(S)在受体序列中的位置。 以马来酰亚胺基二丁酰基生物细胞素(MBB)为底物，进行硫醇生物素化反应. 胰岛素受体硫醇反应性指数。中国仓鼠卵巢细胞 转染人胰岛素受体(CHO/HIRc)细胞 在缺乏或存在谷胱甘肽的情况下耗尽细胞内谷胱甘肽水平 由外源添加的过氧化氢造成的氧化应激。我们 已经发现在细胞氧化的条件下，枯竭的 谷胱甘肽导致受体硫醇反应性的急剧丧失(S)。 在分离和鉴定一种独特的 含有硫醇生物素化残基的胰岛素受体片段。

The purpose of this project is two fold: (1) to assess the regulation of reactive thiol(s) located in the cytoplasmic domain of the insulin receptor by agents that alter glutathione redox cycle; and (2) to perform fragmentation of thiol-biotinylated insulin receptors to determine the location of reactive thiol(s) within the receptor's sequence. Thiol-biotinylation by maleimidodibutyryl biocytin (MBB) was used as an index of insulin receptor thiol reactivity. Chinese hamster ovary cells transfected with the human insulin receptors (CHO/HIRc) were subjected to depletion of cellular glutathione level in the absence or presence of oxidative stress created by exogenous addition of hydrogen peroxide. We have found that under condition of cellular oxidation, depletion of glutathione led to a drastic loss of reactivity of the receptor thiol(s). Progress has been made in the isolation and characterization of a unique insulin receptor fragment containing thiol-biotinylated residue.