Requirement for an accessory factor for binding of [3H]estradiol to protein in the cytosol fraction of rat pancreas.
Requirement for an accessory factor for binding of [3H]estradiol to protein in the cytosol fraction of rat pancreas.
复制标题
[3H]雌二醇与大鼠胰腺细胞质部分中的蛋白质结合需要辅助因子。
DOI：
--
复制
发表时间：
1981
影响因子：
11.1
通讯作者：
A. Grossman
中科院分区：
文献类型：
--
作者：
A. Boctor;P. Band;A. Grossman
当与 5 nM [3H]雌二醇在室温下孵育 1 小时时，来自大鼠胰腺的上清液级分每毫克蛋白质结合约 300 fmol [3H]雌二醇。通过凝胶过滤柱将洗脱液中的结合减少至其初始活性的大约 1%。上清液提取物含有重新激活凝胶滤液中结合的因子。向级分洗脱液中添加辅助因子会产生一个尖锐的活性峰。由于不能重新激活的级分含有与可以重新激活的级分一样多或更多的蛋白质，因此得出结论，辅助因子和[3H]雌二醇结合蛋白的相互作用是特异性的。肽如止痛剂[(1-羧基-2-苯乙基)氨基甲酰基-L-精氨酰-L-缬氨酰-L-精醛]，特别是N-苯甲酰-L-苯丙氨酰-L-缬氨酰-L-精氨酸-对硝基苯胺也增强[3H]雌二醇的结合。辅助因子具有水溶性、可透析性和热稳定性。尽管目前纯化的它含有多种物质，但数据表明辅助因子的至少一种成分是寡肽。
Supernatant fractions from rat pancreas bind approximately 300 fmol of [3H]estradiol per mg of protein when incubated with 5 nM [3H]estradiol for 1 hr at room temperature. Passage through gel filtration columns reduces binding in the eluate to approximately 1% of its initial activity. Extracts of the supernatant contain a factor that reactivates binding in gel filtrates. Addition of accessory factor to fractional eluates gives one sharp peak of activity. Since fractions that cannot be reactivated contain as much or more protein as fractions that can be reactivated, it is concluded that interaction of accessory factor and [3H]estradiol-binding protein is specific. Peptides such as antipain [(1-carboxy-2-phenylethyl)carbamoyl-L-arginyl-L-valyl-L-argininal] and, especially, N-benzoyl-L-phenylalanyl-L-valyl-L-arginine-p-nitroanilide also enhanced binding of [3H]estradiol. Accessory factor is water soluble, dialyzable, and heat stable. Although as currently purified, it contains several substances, the data suggest that at least one component of accessory factor is an oligopeptide.