A reappraisal of the binding characteristics of human thyroxine-binding globulin for 3,5,3'-triiodothyronine and thyroxine.
The binding characteristics of T4 and T3 to dilute plasma were studied separately in five normal euthyroid subjects with normal levels of thyroxine-binding globulin (TBG). Scatchard analyses of these data revealed similar mean affinity constants for T4 [2.0 +/- 0.7 (SD) X 10(9) M-1] and T3 (2.0 +/- 0.7 X 10(9) M-1), but a 5-fold higher capacity for T4 (0.75 +/- 0.18 mol T4/mol TBG) than for T3 (0.14 +/- 0.06 mol T3/mol TBG). Similar results were obtained using various assay buffers, pH concentrations, or separation methods. This characteristic pattern of T4 and T3 binding was retained by thyroid hormone free plasma, with the only difference being a slight parallel shift to the left of the Scatchard plots for both T4 and T3. The calculated affinities (Ka) for T4 and T3 were 5.2 X 10(9) M-1 and 5.2 X 10(9) M-1, respectively. High affinity T4 and T3 binding was abolished in plasma selectively depleted of TBG, but was retained after selective depletion of either prealbumin or albumin. Highly purified TBG, prepared from normal serum, demonstrated binding characteristics for T3 and T4 similar to dilute plasma. Displacement of [125I]T4 from dilute plasma by unlabeled T3 or T4 revealed a binding potency of T3 relative to T4 of 9%. Binding affinities derived from analog displacement studies appear invalid as these calculations assume equal binding capacities of TBG for T4 and T3. It seems clear from these studies, that the binding characteristics of human TBG are inconsistent with a single competitive binding site for thyroid hormones.
D001345:Autoradiography; D001665:Binding Sites; D001667:Binding, Competitive; D004587:Electrophoresis, Agar Gel; D006801:Humans; D011228:Prealbumin / Q000378:metabolism; D011485:Protein Binding; D012709:Serum Albumin / Q000378:metabolism; D013974:Thyroxine / Q000097:blood*; D013975:Thyroxine-Binding Proteins / Q000378:metabolism*; D014284:Triiodothyronine / Q000097:blood*
D011228:Prealbumin; D012709:Serum Albumin; D013975:Thyroxine-Binding Proteins; D014284:Triiodothyronine; D013974:Thyroxine