title: Interaction of thyroid hormone nuclear receptor with antibody: characterization of the thyroid hormone binding site.
abstract: To understand the structural basis in the hormone-dependent transcriptional regulation of human beta 1 thyroid hormone receptor (h-TR beta 1), we characterized the region which interacted with the thyroid hormone, 3,3',5-triiodo-L-thyronine (T3). Using the hormone binding domain of h-TR beta 1 (K206-D461) as an immunogen, we screened for monoclonal antibodies which inhibited the binding of T3 to h-TR beta 1. mAb C3, which recognized native h-TR beta 1, was obtained. Analyses of the binding data indicate that binding of T3 to h-TR beta 1 was competitively inhibited by mAb C3. Using a series of truncated mutants of h-TR beta 1 and synthetic peptides, we mapped the binding site of mAb C3 to the region of E248-V256. Thus, part of T3 binding site in h-TR beta 1 is in this nine-amino acid segment, which was shown by circular dichroism spectroscopy to be a random coil. Based on the proposed model of the hormone binding domain as an alpha/beta barrel, E248-V256 contains part of Loop 1 which is on the same side of the DNA binding domain. These results raise the possibility that Loop 1 could be in direct contact with the nearby DNA binding domain to affect the interaction of DNA binding domain with the T3 target genes.
authors.affiliation: Laboratory of Molecular Biology, DCBDC, National Cancer Institute, Bethesda, MD 20892, USA.
authors.forename: M K
authors.initials: MK
authors.lastname: Bhat
authors.forename: P
authors.initials: P
authors.lastname: McPhie
authors.forename: S Y
authors.initials: SY
authors.lastname: Cheng
mesh_terms: D000595:Amino Acid Sequence; D000911:Antibodies, Monoclonal / Q000276:immunology; D001665:Binding Sites; D000939:Epitopes / Q000276:immunology; D006801:Humans; D008969:Molecular Sequence Data; D011988:Receptors, Thyroid Hormone / Q000378:metabolism*; D014284:Triiodothyronine / Q000276:immunology / Q000378:metabolism
chemical_list: D000911:Antibodies, Monoclonal; D000939:Epitopes; D011988:Receptors, Thyroid Hormone; D014284:Triiodothyronine