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Research on the Mechanism of thyroid hormone action

甲状腺激素作用机制的研究

基本信息

批准号：
62570511
负责人：
NAKAMURA Hirotoshi
金额：
$ 1.09万
依托单位：
Kyoto University
依托单位国家：
日本
项目类别：
Grant-in-Aid for General Scientific Research (C)
财政年份：
1987
资助国家：
日本
起止时间：
1987 至 1988
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-62570511/
关键词：
Thyroid hormone Thyroid hormone receptor Hormone action C-erb A Receptor antibody 甲状腺細胞 c-erb Aレセプター抗体 c-erbA FRT細胞レセプター数の変動

项目摘要

1. Production of an antibody against c-erb A/nuclear T_3 receptor proteins and analysis of nuclear T_3 receptorsIt has been reported that c-erb A encodes nuclear T_3 receptors (NT_3R). Based on the sequence of c-erb A cDNA, we synthesized a polypeptide consisting of 15 amino acids, the sequence of which has high homology between c-erb A _1, and , and raised an anti c-erb A antibody. We have studied the interaction between NT_3R and the anti c-erb A antibody and obtained the following results.(1) The antibody immunoprecipitated NT_3R prepared from rat liver, kidney, brain and human kidney similarly.(2) The antibody specifically inhibited T_3-binding to NT_3R in a competitive manner. It decreased the value for T3-binding association constant dose dependently, while maximal binding capacity was not influenced.(3) No effect of the antibody was detected on T_3-binding to rat liver cytosolic binding proteins or human serum TBG, suggesting that the protein conformation of the T_3-binding region of these proteins might be different from that of NT_3R.(4) The immunoabsorption study revealed that reverse T_3 binding protein was not the same as NT_3R.(5)The antibody immunoprecipitated the in vitro transcription/translation products of c-erb A _1 and .2. Nuclear T_3 receptors in thyroid cells.We have found that nuclear T_3 receptor proteins exist in porcine thyroid cells and cultured rat thyroid cell line, FRTL-5 cells, and that TSH significantly decreases NT_3R level in FRTL-5 cells. The results may suggest the existence of another control system of the hypothalamic-pituitary-thyroid axis via modulation of the hormone receptor levels.

1.抗c-erb A/核T_3受体蛋白抗体的制备及核T_3受体的分析已有报道c-erb A编码核T_3受体(NT_3R)。根据c-erb A基因的序列，我们合成了一个由15个氨基酸组成的多肽，其序列与c-erb A_1高度同源，并产生了抗c-erb A的抗体。我们研究了NT_3R与抗c-erb A抗体的相互作用，得到以下结果：(1)抗体免疫沉淀大鼠肝、肾、脑和人肾的NT_3R：(2)抗体以竞争方式特异性地抑制T_3与NT_3R的结合。(3)抗体对大鼠肝细胞浆结合蛋白和人血清总胆红素结合无影响，提示这些蛋白的T_3结合区的蛋白质构象可能与NT_3R不同。(4)免疫吸收实验表明，反向T_3结合蛋白与NT_3R不同。(5)抗体免疫沉淀c-erb A_1和2的体外转录/翻译产物。在猪甲状腺细胞和培养的大鼠甲状腺细胞系FRTL-5细胞中存在核T_3受体蛋白，TSH可显著降低FRTL-5细胞中NT_3R的水平。这一结果可能表明，通过调节激素受体水平，下丘脑-垂体-甲状腺轴存在另一种控制系统。