Reese J C, Callard I P
Department of Biology, Boston University, MA 02215.
J Steroid Biochem. 1989 Aug;33(2):297-310. doi: 10.1016/0022-4731(89)90308-7.
Two progesterone receptors in the oviduct of the freshwater turtle Chrysemys picta: possible homology to mammalian and avian receptor systems. Here we report the characterization of two specific progesterone receptors in nuclear extracts of the turtle oviduct. The receptors differ in dissociation constants (2.8 nM vs 27 nM) which can be separated on DEAE-Sepharose, the former eluting at 0.08 M KCl and the latter at 0.20 M KCl. [3H]R5020 photoaffinity labeling SDS-PAGE revealed that the 2.8 nM moiety migrates with an apparent molecular weight of 80 +/- 5 kDa and the 27 nM moiety migrates with an apparent molecular weight of 120 +/- 5 kDa. These receptors are termed PR-A and PR-B due to their molecular mass and elution profiles. DNA-cellulose chromatographic studies show that both bind DNA-cellulose with the PR-A eluting at 0.09 M NaCl and PR-B eluting between 0.20-0.21 M NaCl. In reproductively inactive turtles (from the months of January and February) estradiol is undetectable, and PR-B is absent as determined by Scatchard analysis, [3H]R5020 photoaffinity labeling electrophoretic studies and DEAE-Sepharose and DNA-cellulose chromatography. In these animals PR-B can be replenished by estrogen treatment, suggesting a physiological role for both PR-A and PR-B and dependence of PR-B on estradiol.
可能与哺乳动物和鸟类受体系统存在同源性。在此,我们报告了龟输卵管核提取物中两种特异性孕酮受体的特性。这两种受体的解离常数不同(2.8 nM对27 nM),可在DEAE - 琼脂糖上分离,前者在0.08 M KCl浓度下洗脱,后者在0.20 M KCl浓度下洗脱。[3H]R5020光亲和标记SDS - PAGE显示,2.8 nM部分的表观分子量为80±5 kDa,27 nM部分的表观分子量为120±5 kDa。由于它们的分子量和洗脱图谱,这些受体被称为PR - A和PR - B。DNA - 纤维素色谱研究表明,两者均与DNA - 纤维素结合,PR - A在0.09 M NaCl浓度下洗脱,PR - B在0.20 - 0.21 M NaCl浓度之间洗脱。在生殖不活跃的龟(1月和2月)中,通过Scatchard分析、[3H]R5020光亲和标记电泳研究以及DEAE - 琼脂糖和DNA - 纤维素色谱法测定,未检测到雌二醇,且不存在PR - B。在这些动物中,PR - B可通过雌激素处理补充,这表明PR - A和PR - B均具有生理作用,且PR - B依赖于雌二醇。