Bost K L, Blalock J E
Mol Cell Endocrinol. 1986 Jan;44(1):1-9. doi: 10.1016/0303-7207(86)90099-7.
We have used a new methodology to generate a monospecific antiserum to the corticotropin (ACTH) receptor on mouse Y-1 adrenal cells. Using immunoaffinity chromatography the ACTH receptor was purified, and the molecular structure and 125I-ACTH binding characteristics were determined. A molecular weight (Mr) of 225 000 was determined for the complete ACTH receptor as analyzed by sodium dodecyl sulfate (SDS) polyacrylamide gel electrophoresis. The receptor was composed of 4 subunits with Mr 83 000, 64 000, 52 000 and 22 000. The 83 and 52 kDa subunits were disulfide linked and non-covalently associated with the 64 and 22 kDa subunits. The ability to specifically bind 125I-ACTH was localized to the 83 kDa subunit. The purified receptor possessed binding affinities of 3.4 X 10(10) M-1 and 1.0 X 10(9) M-1 as determined by Scatchard analysis.
我们采用了一种新方法来制备针对小鼠Y-1肾上腺细胞促肾上腺皮质激素（ACTH）受体的单特异性抗血清。利用免疫亲和层析法纯化了ACTH受体，并确定了其分子结构和125I-ACTH结合特性。通过十二烷基硫酸钠（SDS）聚丙烯酰胺凝胶电泳分析，完整的ACTH受体分子量（Mr）为225000。该受体由分子量分别为83000、64000、52000和22000的4个亚基组成。83kDa和52kDa的亚基通过二硫键相连，并与64kDa和22kDa的亚基非共价结合。特异性结合125I-ACTH的能力定位于83kDa亚基。通过Scatchard分析确定，纯化后的受体结合亲和力分别为3.4×10(10) M-1和1.0×10(9) M-1。
