Perdew G H, Bradfield C A
Department of Veterinary Science, Pennsylvania State University, University Park 16802, USA.
Biochem Mol Biol Int. 1996 Jun;39(3):589-93. doi: 10.1080/15216549600201651.
Expression of a series of Ah receptor (AhR) deletion mutants in an in vitro translation system has been previously used to map several functional domains of the murine AhR (Dolwick et al. (1993) Proc. Natl. Acad. Sci. USA 90, 8566-8570). In this report, quantitative immunoprecipitation of 90-kDa heat shock protein (hsp90) from reticulocyte lysate allowed us to measure the level of the AhR and AhR deletion mutants complexed with hsp90. After translation of a series of deletion mutants it was determined that there are two distinct domains important in forming a stable AhR/hsp90 complex, corresponding to amino acid sequences 1-166 and 289-347 of the AhR. Neither ARNT, nor Per were able to stably interact with hsp90. Thus, the AhR appears to be a unique member of the PAS domain family of proteins that binds a known ligand and stably interacts with hsp90.
此前，曾利用体外翻译系统表达一系列芳烃受体（AhR）缺失突变体，来定位小鼠AhR的几个功能结构域（Dolwick等人，《美国国家科学院院刊》90, 8566 - 8570（1993年））。在本报告中，通过对网织红细胞裂解物中的90 kDa热休克蛋白（hsp90）进行定量免疫沉淀，我们得以测定与hsp90复合的AhR及AhR缺失突变体的水平。在翻译一系列缺失突变体后，确定在形成稳定的AhR/hsp90复合物方面有两个不同的重要结构域，分别对应AhR的氨基酸序列1 - 166和289 - 347。芳烃核转运蛋白（ARNT）和周期蛋白（Per）均不能与hsp90稳定相互作用。因此，AhR似乎是PAS结构域蛋白家族中的一个独特成员，它能结合已知配体并与hsp90稳定相互作用。
