Denis M, Blanchardie P, Orsonneau J L, Lustenberger P
Department of Medical Biochemistry, Nantes University Hospital, France.
J Chromatogr. 1990 May 25;508(1):97-107. doi: 10.1016/s0021-9673(00)91243-3.
Chromatography of rabbit glucocorticoid-receptor complexes in the absence of sodium molybdate on a Mono Q anion-exchange column induces the transformation of the receptor and allows the resolution of the transformed and non-transformed molecular species. These abilities were used to design a new purification scheme for the glucocorticoid receptor from rabbit liver in its transformed state. Microgram amounts of receptor were obtained using this single-step procedure in less than 2 h. The purification yield was 50-60%. Immunoblot experiments showed that the glucocorticoid receptor was present as an Mr approximately 94,000 polypeptide in these preparations and represented 20-30% of the eluted proteins as determined by densitometric scanning analysis of silver-stained sodium dodecyl sulphate polyacrylamide gels. Finally, the purified receptor was able to interact quantitatively with bulk DNA.
在无钼酸钠的情况下，将兔糖皮质激素受体复合物在Mono Q阴离子交换柱上进行色谱分析，可诱导受体发生转变，并能分离出转变态和非转变态的分子形式。利用这些特性设计了一种从兔肝脏中纯化处于转变态的糖皮质激素受体的新方案。通过这一单步程序，在不到2小时的时间内获得了微克量的受体。纯化产率为50 - 60%。免疫印迹实验表明，在这些制剂中，糖皮质激素受体以分子量约为94,000的多肽形式存在，通过对银染十二烷基硫酸钠聚丙烯酰胺凝胶进行光密度扫描分析测定，其占洗脱蛋白的20 - 30%。最后，纯化后的受体能够与大量DNA进行定量相互作用。
