Galton V A
Endocrinology. 1986 Mar;118(3):1114-8. doi: 10.1210/endo-118-3-1114.
Previous studies have provided conflicting evidence as to whether tadpole liver nuclei contain the same number of high affinity binding sites for T4 as they do for T3. Inability to prepare stable tadpole liver nuclei may have contributed to these inconsistencies. Thus, a study was carried out in which the in vivo binding of T4 and T3 by tadpole liver nuclei was reexamined with a greatly improved method for isolating liver nuclei. It was found that the maximum binding capacities of the nuclei for T3 and T4 were not significantly different [0.161 +/- 0.015 (+/- SEM) vs. 0.185 +/- 0.046 pmol/mg DNA]. However, the affinity of the binding sites for T3 was 2-3 times their affinity for T4 (Kd, 1.65 +/- 0.31 vs. 4.32 +/- 0.92 X 10(-12) M). Furthermore, the nuclear binding of [125I]T4 or [125I]T3 was decreased to comparable levels by administration of saturating amounts of T3 and T4 given either before or with the 125I-labeled hormone, indicating that both hormones were competing for the same set of sites. It is suggested that at least some of the previous conflicting findings related to the number of putative thyroid hormone receptors in tadpole liver nuclei are attributable to inadequate methodology which resulted in an overestimation of the maximum binding capacity for T4.
以往的研究对于蝌蚪肝细胞核中T4的高亲和力结合位点数量是否与T3相同提供了相互矛盾的证据。无法制备稳定的蝌蚪肝细胞核可能导致了这些不一致性。因此,进行了一项研究,用一种大大改进的肝细胞核分离方法重新检查了蝌蚪肝细胞核对T4和T3的体内结合情况。结果发现,细胞核对T3和T4的最大结合能力没有显著差异[0.161±0.015(±标准误)对0.185±0.046 pmol/mg DNA]。然而,结合位点对T3的亲和力是对T4亲和力的2至3倍(解离常数,1.65±0.31对4.32±0.92×10⁻¹² M)。此外,在给予饱和量的T3和T4(在给予¹²⁵I标记激素之前或同时给予)后,[¹²⁵I]T4或[¹²⁵I]T3的核结合降低到了相当的水平,这表明两种激素在竞争同一组位点。有人提出,以前一些关于蝌蚪肝细胞核中假定甲状腺激素受体数量的相互矛盾的发现,至少部分归因于方法不当,导致对T4最大结合能力的高估。