Conformational transition of thyroid hormone receptor upon hormone binding: demonstration by aqueous two-phase partitioning.
An aqueous two-phase partitioning study of partially purified nuclear thyroid hormone receptor from rat liver was performed. Stability of 3,5,3'-tri-iodo-L-thyronine (T3)-receptor complex and T3-binding activity in the presence of dextran or polyethylene glycol were assessed in order to determine the amount of occupied or unoccupied receptors in each phase. Partition coefficients were calculated as the ratio of receptor concentration in the upper polyethylene glycol-rich phase H2O and that in the lower dextran-rich phase H2O. The partition coefficient was a sensitive function of the salt at pH above 6.1 and below 5.1. The salt had no effect on the partition coefficient at pH around 5.6. These results suggest that the isoelectric point of the thyroid hormone receptor is about 5.6, confirming previous determinations using isoelectric focusing. The partition coefficient of the receptor decreased upon T3 binding, regardless of the salt composition. In contrast, the partition coefficient of thyroxine-binding globulin increased upon T3 binding. Free T3 preferentially partitioned into the upper polyethylene glycol-rich phase and gave a partition coefficient higher than 1.0. These results strongly suggest that the decrease in the partition coefficient of the receptor upon hormone binding reflects conformational changes or changes in electrostatic properties of the receptor upon hormone binding. Such an alteration may be involved in biological activation of the receptor upon hormone binding.
Ichikawa
K
K
Department of Gerontology, Endocrinology and Metabolism, Shinshu University School of Medicine, Matsumoto, Japan.
Hashizume
K
K
Miyamoto
T
T
Nishii
Y
Y
Yamauchi
K
K
Ohtsuka
H
H
Yamada
T
T
D000818:Animals; D004247:DNA / Q000378:metabolism; D003911:Dextrans; D006801:Humans; D006863:Hydrogen-Ion Concentration; D011092:Polyethylene Glycols; D011487:Protein Conformation; D051381:Rats; D011988:Receptors, Thyroid Hormone / Q000378:metabolism*; D012492:Salts; D013975:Thyroxine-Binding Proteins; D014284:Triiodothyronine / Q000378:metabolism*
D003911:Dextrans; D011988:Receptors, Thyroid Hormone; D012492:Salts; D013975:Thyroxine-Binding Proteins; D014284:Triiodothyronine; D011092:Polyethylene Glycols; D004247:DNA