Nemoto T, Ohara-Nemoto Y, Ota M
Department of Biochemistry, Iwate Medical University School of Dentistry.
J Biochem. 1987 Sep;102(3):513-23. doi: 10.1093/oxfordjournals.jbchem.a122083.
The nontransformed glucocorticoid receptor (GR) and an 88-kDa protein in rat liver cytosol were selectively adsorbed on protamine- and arginine-Sepharose. The 88-kDa protein was purified from rat liver cytosol to homogeneity by precipitation with protamine sulfate, followed by DEAE-ion exchange chromatography, gel chromatography, and DEAE ion-exchange high-performance liquid chromatography. The 88-kDa protein appeared to be present as a dimer at both low and high salt concentrations. The physicochemical properties and the amino acid composition of the 88-kDa protein were almost the same as those of heat-shock protein 90 of HeLa cells and yeast. Preincubation of the GR with the polyclonal antibody raised against the 88-kDa protein increased the sedimentation coefficient of the nontransformed GR, but did not change that of the transformed GR. These results indicate that heat-shock protein 90 is associated with rat liver nontransformed GR and is responsible for the interaction of GR with protamine.
未转化的糖皮质激素受体（GR）和大鼠肝细胞溶质中的一种88 kDa蛋白被选择性吸附在鱼精蛋白和精氨酸-琼脂糖上。通过硫酸鱼精蛋白沉淀，随后进行DEAE离子交换色谱、凝胶色谱和DEAE离子交换高效液相色谱，从大鼠肝细胞溶质中纯化出88 kDa蛋白至均一性。88 kDa蛋白在低盐和高盐浓度下均以二聚体形式存在。88 kDa蛋白的物理化学性质和氨基酸组成与HeLa细胞和酵母的热休克蛋白90几乎相同。用针对88 kDa蛋白产生的多克隆抗体对GR进行预孵育，增加了未转化GR的沉降系数，但未改变转化GR的沉降系数。这些结果表明，热休克蛋白90与大鼠肝脏未转化的GR相关，并负责GR与鱼精蛋白的相互作用。
