Enomoto M, Kinoshita A, Pan H O, Suzuki F, Yamamoto I, Takigawa M
Department of Biochemistry and Calcified-Tissue Metabolism, Faculty of Dentistry, Osaka University, Japan.
Biochem Biophys Res Commun. 1989 Aug 15;162(3):1222-9. doi: 10.1016/0006-291x(89)90804-8.
Parathyroid hormone (PTH) receptors on cultured rabbit costal chondrocytes were demonstrated using HPLC-purified, radioiodinated [Nle8,-Nle18, Tyr34] bovine PTH-(1-34)amide. PTH binding was found to be specific for PTH agonists and antagonists and dependent on the time and temperature of incubation. Both growth cartilage (GC) cells and resting cartilage (RC) cells were shown to have a single class of saturable, high affinity PTH binding sites with a dissociation constant of 0.6-0.7 nM. However, the numbers of receptors per cell were approximately 49,000 on GC cells and 19,000 on RC cells. After crosslinking the receptors on these cells with the radioligand, one, major 125I-labeled band of 76 kDa was separated by SDS-PAGE.
利用高效液相色谱法纯化的放射性碘化的[异亮氨酸8，异亮氨酸18，酪氨酸34]牛甲状旁腺激素-（1-34）酰胺，证实了培养的兔肋软骨细胞上存在甲状旁腺激素（PTH）受体。发现PTH结合对PTH激动剂和拮抗剂具有特异性，并且依赖于孵育时间和温度。生长软骨（GC）细胞和静止软骨（RC）细胞均显示具有一类单一的可饱和、高亲和力PTH结合位点，解离常数为0.6 - 0.7 nM。然而，每个细胞上的受体数量在GC细胞上约为49,000个，在RC细胞上约为19,000个。在用放射性配体交联这些细胞上的受体后，通过SDS-PAGE分离出一条主要的76 kDa的125I标记带。
