Ekiko D B, Kohail H M, Young J D, Bashirelahi N
Urology. 1986 Jan;27(1):41-8. doi: 10.1016/0090-4295(86)90204-9.
Using glycerol density gradient centrifugation technique, single saturation dose assay, and Scatchard plot, the effect of dextran-coated charcoal (DCC) treatment of homogenate or crude cytosol on estradiol binding protein in human benign prostatic hyperplasia was investigated. Receptor binding is increased after thirty minutes DCC treatment of homogenate or cytosol. Increase in estradiol binding is accompanied by loss in cytosolic protein. A 75 per cent increase in binding of estradiol to its receptor was observed after two hours incubation of DCC with homogenate. The concomitant increase in estradiol binding and decrease in protein concentration in both homogenate or cytosol after DCC treatment indicate the possible removal of some protein(s) which inactivate(s) the estradiol receptor. Removal of cofactors required for activation of proteases and removal of endogenous steroids which could be occupying the estradiol sites also are possible. This simple experimental procedure has improved significantly the methodology for the measurement and characterization of estrogen receptor in human BPH.
采用甘油密度梯度离心技术、单次饱和剂量测定法和Scatchard作图法,研究了葡聚糖包被活性炭(DCC)处理人良性前列腺增生组织匀浆或粗制胞质溶胶对雌二醇结合蛋白的影响。用DCC处理匀浆或胞质溶胶30分钟后,受体结合增加。雌二醇结合增加的同时,胞质蛋白减少。DCC与匀浆孵育2小时后,观察到雌二醇与其受体的结合增加了75%。DCC处理后,匀浆或胞质溶胶中雌二醇结合增加而蛋白质浓度降低,这表明可能去除了一些使雌二醇受体失活的蛋白质。去除蛋白酶激活所需的辅因子以及去除可能占据雌二醇位点的内源性类固醇也是有可能的。这个简单的实验程序显著改进了人良性前列腺增生中雌激素受体的测量和表征方法。