Idziorek T, Sablonniere B, Formstecher P, Dumur V, Dautrevaux M
J Steroid Biochem. 1985 Nov;23(5A):593-7. doi: 10.1016/0022-4731(85)90009-3.
Highly-purified non-transformed rat liver [3H]triamcinolone acetonide-receptor complex was shown to be covalently adsorbed on activated thiol sepharose 4B, a reactive sulfhydryl matrice. Elution by mercaptoethanol in excess and inhibition of binding by previous treatment of the complex with N-ethylmaleimide clearly demonstrated the specificity of the binding by thiol disulfide interchange. The transformed [3H]triamcinolone acetonide-receptor complex, partially purified by DNA-cellulose chromatography, was also retained on activated thiol sepharose 4B. The physicochemical characteristics of both the transformed and non-transformed glucocorticoid receptor complexes eluted from the covalent chromatography column were studied by HP size exclusion chromatography on a TSK G 3000 SW column and were found to be identical to those of the starting complexes. These results provide direct evidence for accessible sulfhydryl groups on the glucocorticoid receptor complex surface, probably distinct from the steroid binding essential sulfhydryl group.
高纯度非转化大鼠肝脏[3H]曲安奈德受体复合物被证明可共价吸附在活化硫醇琼脂糖4B（一种反应性巯基基质）上。用过量的巯基乙醇洗脱以及用N - 乙基马来酰亚胺预先处理复合物对结合的抑制，清楚地证明了通过硫醇 - 二硫键交换进行结合的特异性。通过DNA - 纤维素色谱部分纯化的转化[3H]曲安奈德受体复合物也保留在活化硫醇琼脂糖4B上。在TSK G 3000 SW柱上通过高效尺寸排阻色谱研究了从共价色谱柱洗脱的转化和未转化糖皮质激素受体复合物的物理化学特性，发现它们与起始复合物的特性相同。这些结果为糖皮质激素受体复合物表面存在可及的巯基提供了直接证据，这些巯基可能与类固醇结合必需的巯基不同。
