Gehrke L, Bast R E, Ilan J
J Biol Chem. 1981 Mar 10;256(5):2514-21.
Average rates of polypeptide chain elongation have been determined in cockerel liver explants on 4 successive days following an in vivo injection of 17 beta-estradiol. Incorporation of [3H]leucine by the explants is linear for at least 24 h, and the rate of protein synthesis increases significantly after estrogen injection. The explants synthesize and secrete serum albumin. B-apolipoprotein, and the phosphoprotein vitellogenin at relative rates which are similar to those reported for liver in vivo. Using this system, changes in the average rates of polypeptide chain elongation have been analyzed as a temporal sequence following a single injection of 17 beta-estradiol into cockerels. For this, average ribosome half-transit times were determined by measuring the kinetics of transfer of labeled polypeptides from polysomal-bound to released polypeptides. The data revealed a dramatic effect of estradiol on the average ribosome half-transit time with a maximum increase of 4.6-fold; however, the average size of polypeptides synthesized by explants at the peak of induction increased only 15% when compared to uninduced liver explants. These findings indicate that injection of estradiol results in large changes in the actual rates at which amino acids are added to the growing nascent polypeptide chain; that is, rates of polypeptide chain elongation. Therefore, translation-level regulation of protein synthesis in cockerel livers plays a significant part in determining the magnitude of the response to hormone stimulation.
在给公鸡体内注射17β-雌二醇后的连续4天里,测定了公鸡肝脏外植体中多肽链的平均延伸速率。外植体对[3H]亮氨酸的掺入至少24小时呈线性,雌激素注射后蛋白质合成速率显著增加。外植体合成并分泌血清白蛋白、β-载脂蛋白和磷蛋白卵黄生成素,其相对速率与体内肝脏报道的相似。利用该系统,分析了给公鸡单次注射17β-雌二醇后多肽链平均延伸速率随时间的变化序列。为此,通过测量标记多肽从多核糖体结合态到释放态的转移动力学来确定平均核糖体半转运时间。数据显示,雌二醇对平均核糖体半转运时间有显著影响,最大增加4.6倍;然而,与未诱导的肝脏外植体相比,诱导高峰期外植体合成的多肽平均大小仅增加了15%。这些发现表明,注射雌二醇会导致氨基酸添加到正在生长的新生多肽链的实际速率发生巨大变化,即多肽链延伸速率。因此,公鸡肝脏中蛋白质合成的翻译水平调控在决定对激素刺激的反应程度方面起着重要作用。