Purification and characterization of thyroid-hormone-binding protein from masu salmon serum. A homolog of higher-vertebrate transthyretin.
We purified a thyroid-hormone-binding protein (THBP) from serum of masu salmon at the stage of smoltification when the concentrations of endogenous thyroid hormones in plasma reach the highest levels. All steps of sequential column chromatography suggest that this THBP is responsible for most L-3,5,3'-triiodothyronine-binding activity in serum at this stage. The molecular mass of this protein was estimated to be 60 kDa by gel filtration but only 15 kDa by SDS/PAGE, which suggests that it is comprised of four identical subunits. The amino acid sequence of its N-terminal portion was highly similar to those of vertebrate transthyretins. These molecular features indicate that masu salmon THBP is a homolog of transthyretins from tetrapods. However, in contrast with mammalian transthyretins, the affinity of masu salmon transthyretin for L-3,5,3'-triiodothyronine was three times greater than for L-thyroxine. This rank order affinity is similar to that of avian and frog transthyretins. Scatchard analysis revealed that masu salmon transthyretin possesses a single class of binding site for L-3,5,3'-triiodothyronine, with a Kd of 13.8 nM at 0 degrees C. Taken together with the data reported by Chang et al. [Eur. J. Biochem. (1999) 259, 534-542], these results suggest that transthyretin has changed from a L-3,5, 3'-triiodothyronine-carrier protein to a L-thyroxine-carrier protein during mammalian evolution.
Yamauchi
K
K
Department of Bioloy, Faculty of Science, Shizuoka University, Japan.
Nakajima
J
J
Hayashi
H
H
Hara
A
A
D000595:Amino Acid Sequence; D000818:Animals; D001665:Binding Sites; D001798:Blood Proteins / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification*; D002352:Carrier Proteins / Q000097:blood / Q000235:genetics / Q000302:isolation & purification*; D006801:Humans; D007700:Kinetics; D008565:Membrane Proteins / Q000097:blood / Q000235:genetics / Q000302:isolation & purification*; D008969:Molecular Sequence Data; D008970:Molecular Weight; D011228:Prealbumin / Q000235:genetics; D020836:Protein Structure, Quaternary; D012474:Salmon / Q000097:blood* / Q000235:genetics / Q000254:growth & development; D017386:Sequence Homology, Amino Acid; D013045:Species Specificity; D013963:Thyroid Hormones; D014284:Triiodothyronine / Q000378:metabolism; D000097810:Thyroid Hormone-Binding Proteins
D001798:Blood Proteins; D002352:Carrier Proteins; D008565:Membrane Proteins; D011228:Prealbumin; D013963:Thyroid Hormones; D014284:Triiodothyronine; D000097810:Thyroid Hormone-Binding Proteins