Estrogen receptor: Loss of DNA binding ability following trypsin or Ca2+ treatment
Estrogen receptor: Loss of DNA binding ability following trypsin or Ca2+ treatment
复制标题
雌激素受体：胰蛋白酶或 Ca2 处理后 DNA 结合能力丧失
DOI：
10.1016/0014-5793(73)80866-x
复制
发表时间：
1973
期刊：
影响因子：
3.5
通讯作者：
H. Rochefort
中科院分区：
文献类型：
--
作者：
J. André;H. Rochefort
2.方法取小牛子宫内膜细胞，用[5，7- 3 H]-雌二醇（CEA; spec '，~ e acti~ d~ y= 48 C5] mmole）于0 ℃孵育90 min。然后将1 t与小牛胸腺DNA类型t Sigma L（1.0 pg] ml）在2下混合2 h。将样品分别离心，在105，000 g下通过5-20%蔗糖梯度离心14小时。在α-烯烃锌注射液mL~ t~ r~ 3中计算各组分的放射性（5，0~，效率）和2，6，8 nm吸收艾德法测定的DNA浓度。在未结合的ReE 2区域中的有效性~ 6]。在0 ~（-4）~（-9）℃时，用右旋糖酐修饰的活性炭吸附法测定了特异性吸附位点的数目。蛋白质浓度根据I. owa-y tee~ miqne~ 12].
2. MethodsThe calf uterine cytom] prepared as d~ scrib~ d previously was~ cubated at 0 for 90 rnin with [5, 7-3H]-estradiol (CEA; spec',~ e acti~ d~ y= 48 C5] mmole)'. 1t was then mixed with calf thymus DNA type t Sigma L (]• 0 pg] ml) at 2 for 2 hi. Th~ samples were sepalately centrifuged, flarough a 5-20% sucrose gradient at 105,000 g for 14 bx. The fractions were counted for radioactivity in a~ olaene zcinti]] ation mL~ t~ r~ 3 (5, 0~, efficiency) and 1he DNA concentration-~ valuat, ed by abzorption at 2, 6', 8 nm, The degree, of ahe DNA--R¢ E 2 interned, on was measured not, only by the increase~ f radioactivi, W in the DNA re, on bu~~ s~ by the decrease of radioa, efivJty in the unbound ReE 2 region~ 6]. The number, of specific estradJot bha, ding sites wxis measured by, dSfferential adsorption on dextran-eoated charcoal at 0--4-~[9]. Protein concentration was msayed accordiu~ t, o its absorption~ at 28, 0 and 260 nm mad by the I. owa-y tee~ miqne~ 12].