Yamauchi K, Tata J R
Laboratory of Developmental Biochemistry, National Institute for Medical Research, London, England.
Eur J Biochem. 1994 Nov 1;225(3):1105-12. doi: 10.1111/j.1432-1033.1994.1105b.x.
A variety of cytosolic thyroid-hormone-binding proteins with different characteristics have previously been reported. Here, we first describe the thyroid-hormone-binding characteristics of adult Xenopus liver cytosol, then a novel procedure for purifying cytosolic thyroid-hormone-binding protein (CTBP) from Xenopus liver (xCTBP). The procedure consists of combining preparative isoelectrofocusing, FPLC cation-exchange chromatography, HPLC hydrophobic-interaction chromatography and ultraviolet light cross-linking of 125I-labeled 3,3'5-triiodo-L-thyronine (T3). The isolated xCTBP thus prepared retained all the characteristics of the major thyroid- hormone-(TH)-binding component of the unfractionated cytosol. It is a monomeric protein of approximately 59 kDa with an isoelectric point of 7.0 +/- 0.1, binds T3 with a higher affinity than its analogs with a Kd of approximately 9 nM, and is sensitive to sulfhydryl agents but not to NADPH. In several respects, xCTBP differs from most CTBP-like preparations from other sources described hitherto. Microsequencing of a 23-amino-acid peptide generated from xCTBP by cyanogen bromide digestion revealed 92-100% identity of a 23-amino-acid sequence of several mammalian (amino acids 236-258) and avian (amino acids 245-267) cytosolic aldehyde dehydrogenases (ALDH); xCTBP also exhibited significant similarity of amino acid composition with rat ALDH. This novel finding of sequence identity between a CTBP and ALDH, and the diversity of CTBPs from different sources, suggest that a variety of cytosolic proteins, depending on the species and tissue, can function as thyroid-hormone-binding proteins.
