Effect of heterodimer partner RXRalpha on PPARgamma activation function-2 helix in solution.
The structural mechanism of allosteric communication between retinoid X receptor (RXR) and its heterodimer partners remains controversial. As a first step towards addressing this question, we report a nuclear magnetic resonance (NMR) study on the GW1929-bound peroxisome proliferator-activated receptor gamma (PPARgamma) ligand-binding domain (LBD) with and without the 9-cis-retinoic acid (9cRA)-bound RXRalpha LBD. Sequence-specific 13C(alpha), 13C(beta), and 13CO resonance assignments have been established for over 95% of the 275 residues in the PPARgamma LBD monomer. The 1HN, 15N, and 13CO chemical shift perturbations induced by the RXRalpha LBD binding are located at not only the heterodimer interface that includes the C-terminal residue Y477 but also residues Y473 and K474 in the activation function-2 (AF-2) helix. This result suggests that 9cRA-bound RXRalpha can affect the PPARgamma AF-2 helix in solution and demonstrates that NMR is a powerful new tool for studying the mechanism of allosteric ligand activation in RXR heterodimers.
Lu
Jianyun
J
Department of Internal Medicine, Washington University School of Medicine, St. Louis, MO 63110, USA. lujy@wustl.edu
Chen
Minghe
M
Stanley
Susan E
SE
Li
Ellen
E
D000077556:Alitretinoin; D000495:Allosteric Site; D000595:Amino Acid Sequence; D001665:Binding Sites; D019281:Dimerization; D015723:Gene Library; D006801:Humans; D008024:Ligands; D008958:Models, Molecular; D008969:Molecular Sequence Data; D019906:Nuclear Magnetic Resonance, Biomolecular; D047495:PPAR gamma / Q000737:chemistry* / Q000378:metabolism*; D011487:Protein Conformation; D047490:Retinoid X Receptor alpha / Q000737:chemistry* / Q000378:metabolism*; D012996:Solutions; D014212:Tretinoin / Q000378:metabolism
D008024:Ligands; D047495:PPAR gamma; D047490:Retinoid X Receptor alpha; D012996:Solutions; D000077556:Alitretinoin; D014212:Tretinoin