Thionamides and arsenite inhibit specific T3 binding to the hepatic nuclear receptor.
Methimazole (MMI) and propylthiouracil (PTU) are widely used for the treatment of Graves' disease. However, no studies have been reported on the action of these drugs on binding of L-triiodothyronine (T3) to the nuclear receptor. T3 receptors of rat liver nuclei, prepared by differential centrifugation, were extracted with 0.4 M KCl and 5 mM dithiothreitol (DTT). In the assessment of T3 binding to the DTT-reduced receptor, the hepatic nuclear extract was chromatographed on Superose 6 to remove DTT and isolate proteins of relative mass approximately 50,000 (chromatographed nuclear receptors (CNRs)), prior to the addition of [125I]T3 of high specific activity (3300 microCi/micrograms; 1 Ci = 37 GBq). MMI or PTU at 2 mM reduced specific T3 binding to CNR by 84% and 85%, respectively. The inhibitory effects of these reagents and 2 mM sodium arsenite (which complexes dithiols) were additive. Scatchard analyses indicated that neither MMI nor PTU (at 2 mM) significantly altered the affinity constant (Ka) (from 2.41 x 10(9) to 1.74 x 10(9) M-1 for PTU and 1.79 x 10(9) M-1 for MMI), while they both decreased (p less than 0.02) maximal binding capacity (from 0.36 +/- 0.02 to 0.19 +/- 0.02 pmol/mg protein for MMI and 0.17 +/- 0.02 pmol/mg protein for PTU). Dose-response curves showed that 50% inhibition was attained at 0.6 mM PTU or 1.0 mM MMI with approximately 25% inhibition by both at 0.1 mM. Artefactual binding effects by MMI and PTU on [125I]T3 were excluded by chromatography experiments. Similar results were obtained using nuclear receptors prepared from livers of hyperthyroid rats.(ABSTRACT TRUNCATED AT 250 WORDS)
Takagi
S
S
Thyroid Research Laboratory, Mount Sinai Hospital Research Institute, Toronto, Ont., Canada.
Hummel
B C
BC
Walfish
P G
PG
D000818:Animals; D001151:Arsenic / Q000494:pharmacology*; D018053:Arsenites; D002467:Cell Nucleus / Q000378:metabolism; D002853:Chromatography, Liquid; D004229:Dithiothreitol / Q000494:pharmacology; D008099:Liver / Q000378:metabolism*; D008713:Methimazole / Q000494:pharmacology*; D008741:Methyl Methanesulfonate / Q000031:analogs & derivatives / Q000037:antagonists & inhibitors / Q000494:pharmacology; D011441:Propylthiouracil / Q000494:pharmacology*; D051381:Rats; D011988:Receptors, Thyroid Hormone / Q000187:drug effects* / Q000378:metabolism; D017670:Sodium Compounds; D013439:Sulfhydryl Reagents / Q000494:pharmacology*; D014284:Triiodothyronine / Q000378:metabolism*
D018053:Arsenites; D011988:Receptors, Thyroid Hormone; D017670:Sodium Compounds; D013439:Sulfhydryl Reagents; D014284:Triiodothyronine; C014674:methyl methanethiosulfonate; C017947:sodium arsenite; D008713:Methimazole; D011441:Propylthiouracil; D008741:Methyl Methanesulfonate; D001151:Arsenic; D004229:Dithiothreitol