Purification and characterization of NADPH-dependent cytosolic 3,5,3'-triiodo-L-thyronine binding protein in rat kidney.
Purification and characterization of NADPH-dependent cytosolic 3,5,3'-triiodo-L-thyronine binding protein in rat kidney.
复制标题
大鼠肾中 NADPH 依赖性胞质 3,5,3-三碘-L-甲状腺氨酸结合蛋白的纯化和表征。
DOI：
10.1016/s0021-9258(18)83670-5
复制
发表时间：
1989
期刊：
影响因子：
--
通讯作者：
Takashi Yamada
中科院分区：
文献类型：
--
作者：
Kiyoshi Hashizume;Takahide Miyamoto;Kazuo Ichikawa;K. Yamauchi;Mutsuhiro Kobayashi;Akihiro Sakurai;Hiromi Ohtsuka;Yutaka Nishii;Takashi Yamada
用活性炭抽提、Mono Q-Sepharose、Blue Sepharose CL-6 B和Sephacryl S-200柱层析，从大鼠肾脏中纯化了NADPH依赖的胞浆3，5，3 ′-三碘-L-甲状腺原氨酸（T3）结合蛋白（CTBP），纯化倍数超过30，000倍。纯化的CTBP具有4.7S的沉降系数，Stokes半径为32.5 μ m，并且计算的分子量为58，000。十二烷基硫酸钠-聚丙烯酰胺凝胶电泳显示，该蛋白质为单一多肽链，分子量为58，000。对T3结合的Scatchard分析表明，NADPH增加了最大结合能力，而亲和力常数不变（Ka= 2.43 × 109 M −1）。NADPH和结合能力的双倒数分析得出CTBP的最大结合能力为16，400 pmol/mg，Mr= 58，000。三碘甲腺原氨酸类似物与CTBP的亲和力大小顺序为：L-T3= D-T3>三碘甲腺乙酸> L-甲状腺素。[125 I] T3结合到在20 °C（t1/2 = 22 min）在不存在NADPH的情况下自发地从CTBP解离的纯化CTBP，而在存在NADPH的情况下未观察到解离。T3与CTBP结合的最适pH为7.2-7.5。Na+、K+、Ca ~（2+）和Mg ~（2+）（0-200 mM）对T3与CTBP的结合无影响。纯化的CTBP不与DNA结合，也不吸附到伴刀豆球蛋白A-Sepharose上。
The NADPH-dependent cytosolic 3,5,3′-triiodo-L-thyronine(T3)-binding protein (CTBP) has been purified over 30,000-fold from rat kidney by using charcoal extraction, Mono Q-Sepharose, Blue Sepharose CL-6B, and Sephacryl S-200 column chromatography. Purified CTBP had a sedimentation coefficient of 4.7 S, Stokes radius of 32.5Å, and calculated molecular weight of 58,000. The apparently homogeneous protein consisted of a single polypeptide chain withMrof 58,000 as estimated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Scatchard analysis of T3binding showed that NADPH increases maximal binding capacity without changes in the affinity constant (Ka= 2.43 × 109M−1). Double reciprocal analysis of NADPH and binding capacity gave maximal binding capacity of 16,400 pmol/mg of CTBP,Mr= 58,000. The order of affinity of iodothyronine analogues to purified CTBP was as follows:L-T3= D-T3> triiodothyroacetic acid >L-thyroxine. [125I]T3bound to purified CTBP spontaneously dissociated from CTBP at 20 °C (t½= 22 min) in the absence of NADPH, whereas the dissociation was not observed in the presence of NADPH. The optimal pH for T3binding was 7.2–7.5 Na+, K+, Ca2+, and Mg2+(0-200 mM) did not influence T3binding to CTBP. The purified CTBP did not bind to DNA and was not adsorbed to concanavalin A-Sepharose.
登录
查看更多内容
NADP 结合蛋白导致人红细胞中 NADP 的可用性和乙状释放减少。
DOI：
--
发表时间：
1986
期刊：
The Journal of biological chemistry
影响因子：
--
作者：
Kirkman,HN;Gaetani,GF;Clemons,EH
通讯作者：
Clemons,EH
通过胰蛋白酶消化大鼠肝核甲状腺激素受体，将 DNA 结合域与激素和核心组蛋白结合域分离。
DOI：
--
发表时间：
1986
期刊：
The Journal of biological chemistry
影响因子：
--
作者：
Ichikawa,K;DeGroot,LJ
通讯作者：
DeGroot,LJ
分离肝细胞中 3,5,3-三碘甲状腺原氨酸的跨细胞和跨核转运。
影响因子：
4.8
作者：
Mooradian,AD;Schwartz,HL;Mariash,CN;Oppenheimer,JH
通讯作者：
Oppenheimer,JH
在大鼠肝、肾、脑和心脏中，三碘甲状腺原氨酸从血浆到细胞质以及从细胞质到细胞核的立体特异性转运。
DOI：
10.1172/jci111667
发表时间：
1985
期刊：
The Journal of clinical investigation
影响因子：
--
作者：
Oppenheimer,JH;Schwartz,HL
通讯作者：
Schwartz,HL