Barlow, J W
De Nayer, Pierre-Pascal
[UCL]
Although putatively identified more than 10 years ago, thyroid hormone receptors in human tissues remain poorly characterized. As a first step towards understanding the mechanism of thyroid hormone action in man we have characterized T3 binding sites in nuclei of the human lymphoblastoid line, IM-9 cells. In whole cell experiments at 37 degrees C, nuclear binding of [125I]T3 was saturable (Kd 34 +/- 6 pmol/l) and of finite capacity (approximately equal to 350 sites/cell). The binding sites were extracted from a nuclear pellet by treatment with 0.4 mol/l KCl and sonication. Separation of bound from free [125I]T3 in the extracts was achieved using the calcium phosphate matrix, hydroxyapatite at a concentration of 0.3 ml of a 150 g/l slurry. Rectilinear Scatchard plots were obtained only when the hydroxyapatite was washed with a buffer containing 0.5% Triton X-100. Under these conditions T3 binding sites in the nuclear extracts were present at a concentration of 22.4 +/- 8.6 fmol/mg protein and showed an affinity of (Kd, room temperature) 140 +/- 10 pmol/l. The same assay system was used to determine the hierarchy of affinities for a range of natural and synthetic analogues. Calling T3 100, the order of potencies observed was: Triac, 500; 3,5-diiodo-3'-isopropylthyronine, 89; T4, 32; 3,5-dimethyl-3'isopropylthyronine 2; 3,5-T2, 0.7, rT3, 0.4; 3'5'-T2, less than 0.01. These results suggest that the T3 binding sites present in human IM-9 lymphocyte nuclei and extracts thereof are thyroid hormone receptors. These cells may be a useful tool to increase our understanding of human T3 receptors.(ABSTRACT TRUNCATED AT 250 WORDS)
Bibliographic reference |
Barlow, J W ; De Nayer, Pierre-Pascal. Characterization of thyroid hormone receptors in human IM-9 lymphocytes.. In: Acta endocrinologica, Vol. 117, no. 3, p. 327-32 (1988) |
Permanent URL |
http://hdl.handle.net/2078.1/29411 |