Cary A J, Martin P A, Dougherty J J
Department of Biochemistry and Biophysics, University of Rhode Island, Kingston 02881.
Arch Biochem Biophys. 1991 Jul;288(1):287-92. doi: 10.1016/0003-9861(91)90197-q.
Following the binding of a specific ligand such as 2,3,7,8-tetrachlorodibenzo-p-dioxin, mild heating has previously been shown to convert cytosolic aryl hydrocarbon (Ah) receptor to two DNA-binding forms: peak I, which appears to be an Ah receptor monomer; and peak II, which is larger and resembles nuclear Ah receptor. In rat liver cytosol pretreated with charcoal-dextran, and heated briefly at 22 degrees C, the addition of a physiological (3 mM) concentration of ATP substantially increases the formation of both peak I and peak II receptor. On more extended incubation in the presence of ATP most of the Ah receptor converts to the tighter binding peak II form. The ATP analog 5'-adenylylmethylene diphosphonate (AMPPCP) stimulates the appearance of both DNA-binding forms as effectively as ATP does. In cytosol separated from low molecular weight components by gel filtration prior to incubation, ATP substantially stimulates the appearance of peak II receptor. ATP also increases the amount of peak II receptor formed when peak I receptor is incubated with unlabeled charcoal-treated cytosol. Thus, ATP promotes both the release of Ah receptor monomer from the 9 S complex which cannot bind DNA and the subsequent conversion of that monomer to a form similar or identical to nuclear Ah receptor.
在特定配体如2,3,7,8 - 四氯二苯并 - p - 二恶英结合后,先前已表明温和加热可将胞质芳烃(Ah)受体转化为两种DNA结合形式:峰I,似乎是Ah受体单体;峰II,更大且类似于核Ah受体。在经活性炭 - 葡聚糖预处理并在22℃短暂加热的大鼠肝脏胞质溶胶中,添加生理浓度(3 mM)的ATP会显著增加峰I和峰II受体的形成。在ATP存在下进行更长时间的孵育,大多数Ah受体转化为结合更紧密的峰II形式。ATP类似物5'-腺苷亚甲基二磷酸(AMPPCP)刺激两种DNA结合形式出现的效果与ATP相同。在孵育前通过凝胶过滤从低分子量成分中分离出的胞质溶胶中,ATP显著刺激峰II受体的出现。当峰I受体与未标记的经活性炭处理的胞质溶胶一起孵育时,ATP也会增加形成的峰II受体的量。因此,ATP既促进了不能结合DNA的9S复合物中Ah受体单体的释放,又促进了该单体随后转化为与核Ah受体相似或相同的形式。