Thyroid hormone action: identification of the mitochondrial thyroid hormone receptor as adenine nucleotide translocase.
A preliminary report from our laboratory suggested that the thyroid hormone triiodothyronine (T3) is bound with an association constant (Ka) approximating 2 x 10(11) M-1 by adenine nucleotide translocase (AdNT) purified from beef heart mitochondria. We now report that [125I]T3 is capable of photoaffinity labeling not only purified AdNT but also the carrier in intact beef heart mitochondria. Photoaffinity labeling in intact mitochondria was appreciably greater than that observed with purified AdNT. The covalently labeled AdNT was identified by 2-dimensional electrophoresis with pI of 10 on electrofocusing and M(r) of 31,000 on SDS gel. Identification of the covalently labeled protein as authentic AdNT was substantiated by its interaction with a specific monoclonal antibody preparation.
Sterling
K
K
Department of Medicine, Columbia University, College of Physicians and Surgeons, New York, New York.
D000818:Animals; D000911:Antibodies, Monoclonal; D002417:Cattle; D015180:Electrophoresis, Gel, Two-Dimensional; D004591:Electrophoresis, Polyacrylamide Gel; D007457:Iodine Radioisotopes; D007525:Isoelectric Focusing; D008929:Mitochondria, Heart / Q000737:chemistry / Q000201:enzymology / Q000648:ultrastructure*; D000226:Mitochondrial ADP, ATP Translocases / Q000032:analysis* / Q000378:metabolism; D011988:Receptors, Thyroid Hormone / Q000032:analysis* / Q000378:metabolism; D014284:Triiodothyronine / Q000032:analysis / Q000378:metabolism
D000911:Antibodies, Monoclonal; D007457:Iodine Radioisotopes; D011988:Receptors, Thyroid Hormone; D014284:Triiodothyronine; D000226:Mitochondrial ADP, ATP Translocases