Gehring U, Segnitz B
Institut für Biologische Chemie, Universität Heidelberg, F.R.G.
Mol Cell Endocrinol. 1988 Apr;56(3):245-54. doi: 10.1016/0303-7207(88)90067-6.
Glucocorticoid receptors of several rodent and human cell lines were subjected to mild proteolysis with several proteases. A hormone binding fragment of Mr approximately 40,000 was generated which had increased affinity for DNA as revealed by DNA-cellulose chromatography. It behaved similar to the truncated nti ('increased nuclear transfer') receptor of mutant mouse lymphoma cells. These data led to the view that wild-type receptors of Mr approximately 94,000 contain in addition to the functional domains for hormone binding and interaction with DNA a third domain ('modulation domain') which is essential for biological activity. Monoclonal antibodies against wild-type receptors were used in DNA binding experiments and increased affinity for DNA was observed. The data suggest that reacting the receptor with antibody leads to functional elimination of the modulation domain as if it were cleaved off by mild proteolysis. Antibody treatment neither caused nor inhibited receptor activation to a DNA binding form.
几种啮齿动物和人类细胞系的糖皮质激素受体用几种蛋白酶进行了温和的蛋白水解。产生了一个分子量约为40,000的激素结合片段，通过DNA纤维素色谱法显示其对DNA的亲和力增加。它的行为类似于突变小鼠淋巴瘤细胞的截短的nti（“增加的核转运”）受体。这些数据导致这样一种观点，即分子量约为94,000的野生型受体除了含有激素结合和与DNA相互作用的功能域外，还含有对生物活性至关重要的第三个结构域（“调节结构域”）。针对野生型受体的单克隆抗体用于DNA结合实验，并观察到对DNA的亲和力增加。数据表明，使受体与抗体反应会导致调节结构域的功能消除，就好像它被温和的蛋白水解切割掉了一样。抗体处理既不会引起也不会抑制受体激活为DNA结合形式。
