Sala-Trepat J M, Hibner U, Vallet-Strouvé C
Nucleic Acids Res. 1977 Mar;4(3):649-62. doi: 10.1093/nar/4.3.649.
The intranuclear distribution of [3H]-estradiol binding sites was studied in highly purified nuclei isolated from calf endometrial tissue pre-incubated with the labeled hormone. The major part (approximately 85%) of the receptor bound estradiol was found associated with the extranucleolar chromatin; only a negligible amount of [3H]-estradiol (approximately 8%) sedimented with the nucleolar fraction. [3H]-estradiol labeled chromatin was then fragmented by sonication and fractionated by sucrose density gradient sedimentation under different conditions of centrifugation. The vast majority of the [3H]-estradiol was invariably found to be associated with a fast sedimenting fraction which contained only 5 to 10% of the nuclear DNA. The concentration of estradiol receptors (per weight of DNA) in this fraction was 25- to 50-fold higher than that found in the slow sedimenting major chromatin component. Chemical analysis showed this fraction to have a high protein/DNA ratio but no phospholipids were detected.
用标记激素预孵育小牛子宫内膜组织后,从其中分离出高度纯化的细胞核,研究了[3H] - 雌二醇结合位点的核内分布。发现与雌二醇结合的受体的主要部分(约85%)与核仁外染色质相关;只有极少量的[3H] - 雌二醇(约8%)与核仁部分一起沉降。然后通过超声处理将[3H] - 雌二醇标记的染色质片段化,并在不同离心条件下通过蔗糖密度梯度沉降进行分级分离。绝大多数[3H] - 雌二醇始终与一个快速沉降的部分相关,该部分仅包含5%至10%的核DNA。该部分中雌二醇受体(每重量DNA)的浓度比缓慢沉降的主要染色质成分中高25至50倍。化学分析表明该部分具有高蛋白/DNA比率,但未检测到磷脂。