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Purification of cytosolic 3,5,3'-triiodo-L-thyronine(T3)-binding protein(CTBP) which regulates nuclear T3
K Hashizume1, S Suzuki, K Ichikawa
1Department of Geriatrics, Endocrinology and Metabolism, Shinshu University School of Medicine, Matsumoto, Japan.
Abstract:
The NADPH-dependent cytosolic 3,5,3'-triiodo-L-thyronine(T3)-binding protein(CTBP) was purified from rat kidney using Mono Q-Sepharose, Red sepharose and T3 affinity chromatography. CTBP which was partially purified by Red Sepharose column chromatography was adsorbed to T3 affinity column in the presence of 50 uM NADPH. The CTBP was eluted from the gel with the buffer which did not contain NADPH. One molecule of the purified CTBP(58 kDa) bound one molecule of T3 with 2.44 x 10(9) M-1 of affinity constant. The purified CTBP was activated not only by NADPH but also by NADP in the presence of dithiothreitol. The NADPH-activated form did not transfer T3 to nuclei, whereas NADP transformed the NADPH-activated CTBP to active form which was able to transfer T3 to nuclei. These results suggested that CTBP-dependent transport of T3 to nucleus is controlled by NADPH and NADP.