Analysis of Agonist and Antagonist Effects on Thyroid Hormone Receptor Conformation by Hydrogen/Deuterium Exchange

Author(s)

Figueira, A. C. M. ^[1] ; Saidemberg, D. M. ^[2] ; Souza, P. C. T. ^[3] ; Martinez, L. ^[3] ; Scanlan, T. S. ^[4] ; Baxter, J. D. ^{[5, 6]} ; Skaf, M. S. ^[3] ; Palma, M. S. ^[2] ; Webb, P. ^{[5, 6]} ; Polikarpov, I. ^[7]

Affiliation

^[1] Lab Nacl Biociencias, BR-13083100 Campinas, SP - Brazil

^[2] Univ Estadual Paulista, Inst Biocencias Rio Claro, Ctr Estudos Insetos Sociais, Dept Biol, BR-13506900 Rio Claro, SP - Brazil

^[3] Univ Estadual Campinas, Inst Chem, BR-13083970 Campinas, SP - Brazil

^[4] Oregon Hlth & Sci Univ, Dept Physiol & Pharmacol, Portland, OR 97201 - USA

^[5] Methodist Hosp, Res Inst, Ctr Diabet, Houston, TX 77030 - USA

^[6] Methodist Hosp, Res Inst, Canc Res Unit, Houston, TX 77030 - USA

^[7] Univ Sao Paulo, Dept Fis & Informat, Inst Fis Sao Carlos, Lab Cristalog Prot, BR-13560970 Sao Carlos, SP - Brazil

• Document type: Journal article
• Source: MOLECULAR ENDOCRINOLOGY; v. 25, n. 1, p. 15-31, JAN 2011.
• Web of Science Citations: 31

Abstract

Thyroid hormone receptors (TRs) are ligand-gated transcription factors with critical roles in development and metabolism. Although x-ray structures of TR ligand-binding domains (LBDs) with agonists are available, comparable structures without ligand (apo-TR) or with antagonists are not. It remains important to understand apo-LBD conformation and the way that it rearranges with ligands to develop better TR pharmaceuticals. In this study, we conducted hydrogen/deuterium exchange on TR LBDs with or without agonist (T(3)) or antagonist (NH(3)). Both ligands reduce deuterium incorporation into LBD amide hydrogens, implying tighter overall folding of the domain. As predicted, mass spectroscopic analysis of individual proteolytic peptides after hydrogen/deuterium exchange reveals that ligand increases the degree of solvent protection of regions close to the buried ligand-binding pocket. However, there is also extensive ligand protection of other regions, including the dimer surface at H10-H11, providing evidence for allosteric communication between the ligand-binding pocket and distant interaction surfaces. Surprisingly, C-terminal activation helix H12, which is known to alter position with ligand, remains relatively protected from solvent in all conditions suggesting that it is packed against the LBD irrespective of the presence or type of ligand. T(3), but not NH(3), increases accessibility of the upper part of H3-H5 to solvent, and we propose that TR H12 interacts with this region in apo-TR and that this interaction is blocked by T(3) but not NH(3.) We present data from site-directed mutagenesis experiments and molecular dynamics simulations that lend support to this structural model of apo-TR and its ligand-dependent conformational changes. (Molecular Endocrinology 25: 15-31, 2011) (AU)

• FAPESP's process: 08/00078-1 - Structure and biophysical studies of complexes of nuclear receptors, ligands, DNA responsive elements and corregulators proteins
• Grantee: Ana Carolina Migliorini Figueira
• Support Opportunities: Scholarships in Brazil - Post-Doctoral
• FAPESP's process: 06/00182-8 - Structural biophysics of nuclear receptors and related proteins
• Grantee: Igor Polikarpov
• Support Opportunities: Research Projects - Thematic Grants