Hutchison K A, Scherrer L C, Czar M J, Stancato L F, Chow Y H, Jove R, Pratt W B
Department of Pharmacology, University of Michigan Medical School, Ann Arbor 48109.
Ann N Y Acad Sci. 1993 Jun 11;684:35-48. doi: 10.1111/j.1749-6632.1993.tb32269.x.
Incubation of immunopurified, hormone-free mouse glucocorticoid receptors with rabbit reticulocyte lysate results in ATP-dependent and monovalent cation-dependent assembly of the GR into a heterocomplex with hsp90, hsp70, and hsp56. Heterocomplex assembly is accompanied by conversion of the receptor from a form that does not bind steroid to a high affinity steroid-binding conformation. Reticulocyte lysate also promotes ATP-dependent dissociation of unliganded receptors from a prebound receptor-DNA complex. Receptor released from DNA has been reconstituted into the heat shock protein heterocomplex and converted to the non-DNA-binding state. The reticulocyte lysate also reconstitutes pp60v-src into a heterocomplex containing hsp90 and p50, both of which are components of the native heterocomplex form of the tyrosine kinase in cytoplasm. Although the c-Raf-1 serine/threonine kinase has never been found in native association with hsp90, it can be assembled into a heat shock protein heterocomplex by the ATP-dependent system in reticulocyte lysate.
将免疫纯化的、无激素的小鼠糖皮质激素受体与兔网织红细胞裂解物一起温育，会导致糖皮质激素受体以ATP依赖和单价阳离子依赖的方式与热休克蛋白90（hsp90）、热休克蛋白70（hsp70）和热休克蛋白56（hsp56）组装成异源复合物。异源复合物的组装伴随着受体从一种不结合类固醇的形式转变为高亲和力的类固醇结合构象。网织红细胞裂解物还能促进未结合配体的受体从预先结合的受体 - DNA复合物中以ATP依赖的方式解离。从DNA上释放的受体已被重新组装成热休克蛋白异源复合物，并转变为非DNA结合状态。网织红细胞裂解物还能将pp60v-src重新组装成一个包含hsp90和p50的异源复合物，这两者都是细胞质中酪氨酸激酶天然异源复合物形式的组成成分。尽管c-Raf-1丝氨酸/苏氨酸激酶从未在天然状态下与hsp90结合，但它可以通过网织红细胞裂解物中的ATP依赖系统组装成热休克蛋白异源复合物。
