Stroupe S D, Westphal U
Biochemistry. 1975 Jul 29;14(15):3296-300. doi: 10.1021/bi00686a002.
An improved purification procedure for the progesterone-binding globulin (PBG) of the pregnant guinea pig has been developed utilizing sulfopropyl Sephadex, a strong cation exchanger, in the first step. The method exploits the low pI (2.8) and favorable acid stability of the glycoprotein. Subsequent chromatographies on DEAE-cellulose and Sephadex G-200 afford a highly purified PBG that exhibits the previously observed polydispersity (R.M. Burton et al. (1974), Biochemistry 13, 3554-3561). Circular dichroism, optical rotatory dispersion, and difference uv spectra all indicate the purified protein to undergo a conformational transition upon forming a complex with a steroid ligand. The CD and ORD spectra cannot be interpreted in terms of tertiary structure probably due to carbohydrate contributions. However, the difference spectra indicate strong perturbation of both a tryptophan residue and the steroid chromophore in the complex.
已经开发出一种改进的纯化怀孕豚鼠孕酮结合球蛋白(PBG)的方法,第一步使用强阳离子交换剂磺丙基葡聚糖凝胶。该方法利用了糖蛋白的低pI(2.8)和良好的酸稳定性。随后在DEAE-纤维素和葡聚糖凝胶G-200上进行色谱分离,得到高度纯化的PBG,其呈现出先前观察到的多分散性(R.M.伯顿等人(1974年),《生物化学》13卷,3554 - 3561页)。圆二色性、旋光色散和差示紫外光谱均表明,纯化后的蛋白质在与类固醇配体形成复合物时会发生构象转变。由于碳水化合物的贡献,CD和ORD光谱可能无法用三级结构来解释。然而,差示光谱表明复合物中的色氨酸残基和类固醇发色团均受到强烈扰动。