Studies on the hormone action at the target tissues: Activation of receptor and metabolism of DNA

靶组织的激素作用研究：受体的激活和DNA的代谢

• 批准号: 61480249
• 负责人: HASHIZUME Kiyoshi
• 金额: $ 3.97万
• 依托单位: Shinshu University School of Medicine
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for General Scientific Research (B)
• 财政年份: 1986
• 资助国家: 日本
• 起止时间: 1986 至 1988
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-61480249/
• 关键词: Cytosolic 3,5,3'-triiodo-L-thyronine(T_3)-binding protein(CTBP); Acceptor protein; nuclear T_3 receptor; NADP; NADPH; Counterregulation; counterregulation; 遣伝子発現調節; 甲状腺ホルモン受容体; 細胞質受容体; 細胞核受容体; 遺伝子発現; 受容体受容蛋白; ADPリボシレーション; 核基質; ホルモン能態輸送; 活性

Cytosolic 3,5,3'-triiodo-l-thyronine(T_3)-binding protein(CTBP) was purified from rat kidney. The molecular weight of CTBP was estimated by gel exclusion study, sucrose density gradient centrifugation and SDS-polyacrylamide gel electrophoresis, and gave a result that MR was 58,000,which was different from that of nuclear T_3 receptor. The purified CTBP was activated by NADPH or by NADP in the presence of -SH residue. The maximal binding capacity was calculated and gave a result that one molecule of CTBP can bind one molecule of T_3. The affinity of T_3 binding to CTBP was identical to that of nuclear T_3 receptor, although nuclear T_3 receptor had a different molecular size.These pyridine nucleotides increased the maximal binding capacity without changes in the affinity constant. NADPH-activated CTBP did not bind to nuclei. However, the NADP-activated CTBP could bind to the nuclei. By extraction with 0.5 M NaCl, acceptor protein for NADP-activated CTBP was isolated. The molecular weight of the acceptor protein was 200,000. NADPH-activated CTBP, therefore, may play a role as a reservoir for intracellular T_3. And NADPH-activated CTBP may play a role in the regulation of gene expression induced by T_3 binding to its specific receptor in nuclei. NADP and NADPH take play as counterregulatory factors for T_3-induced gene expression.

从大鼠肾脏中纯化出胞质3,5,3'-三碘-L-甲状腺氨酸(T_3)结合蛋白(CTBP)。通过凝胶排阻研究、蔗糖密度梯度离心和SDS-聚丙烯酰胺凝胶电泳测定CTBP的分子量，得出MR为58,000，与核T_3受体的分子量不同。纯化的 CTBP 在 -SH 残基存在的情况下被 NADPH 或 NADP 激活。计算最大结合容量，得出一分子CTBP可结合一分子T_3。 T_3与CTBP结合的亲和力与核T_3受体的亲和力相同，尽管核T_3受体具有不同的分子大小。这些吡啶核苷酸增加了最大结合能力而不改变亲和常数。 NADPH 激活的 CTBP 不与细胞核结合。然而，NADP 激活的 CTBP 可以与细胞核结合。通过用 0.5 M NaCl 提取，分离出 NADP 激活的 CTBP 的受体蛋白。受体蛋白的分子量为200,000。因此，NADPH 激活的 CTBP 可能充当细胞内 T_3 的储存库。 NADPH激活的CTBP可能在T_3与其细胞核特异性受体结合诱导的基因表达调控中发挥作用。 NADP 和 NADPH 作为 T_3 诱导基因表达的反调节因子。

项目成果

Hashizume Kiyoshi;Miyamoto Takahide;Nishii Yutaka;Kobayashi Mutsuhiro: Endocrinologia Japonica. 34. 479-487 (1987)

桥爪清；宫本贵秀；西井丰；小林睦宏：日本内分泌学。

Kiyoshi HASHIZUME: "Counterregulation of nuclear 3,5,3'-triiodo-L-thyronine(T3)-binding by oxidized and reduced-nicotinamide adenine dinucleotide phosphates in the presence of cytosolic T3-binding protein in vitro." ENDOCRINOLOGY. 124. (1989)

Kiyoshi HASHIZUME：“体外存在胞质 T3 结合蛋白时，氧化和还原烟酰胺腺嘌呤二核苷酸磷酸对核 3,5,3-三碘-L-甲状腺氨酸 (T3) 结合的反调节。”

Kiyoshi HASHIZUME: "Conformational transition of human c-erb A protein upon thyroid hormone binding" J. Endocrinol.119. 431-437 (1988)

Kiyoshi HASHIZUME：“甲状腺激素结合后人 c-erb A 蛋白的构象转变”J. Endocrinol.119。

橋爪潔志: ENDOCRINOLOGY. 123. 1916-1922 (1989)

桥爪清：内分泌学。123。1916-1922（1989）

Kiyoshi HASHIZUME: "Evidence for the presence of two active forms of cytosolic 3,5,3'-triiodo-L-thyronine(T3)-binding protein(CTBP) in rat kidney. Specialized functions of two CTBPs in intracellular T3 translocation." J. Biol. Chem.264. (1989)

Kiyoshi HASHIZUME：“大鼠肾脏中存在两种活性形式的胞质 3,5,3-三碘-L-甲状腺氨酸 (T3) 结合蛋白 (CTBP) 的证据。两种 CTBP 在细胞内 T3 易位中的特殊功能。”