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Effects of Synthetic Ligands on Heterodimer Pairs Regarding PPAR α , RXR α and LXR α
Author(s) -
Delman Emily Erin,
Rider Stanley Dean,
Hostetler Heather Ann
Publication year - 2016
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.30.1_supplement.630.1
Subject(s) - retinoid x receptor , liver x receptor , nuclear receptor , bimolecular fluorescence complementation , förster resonance energy transfer , receptor , chemistry , retinoid x receptor alpha , microbiology and biotechnology , ligand (biochemistry) , subcellular localization , yellow fluorescent protein , biophysics , transfection , biochemistry , biology , transcription factor , fluorescence , cytoplasm , physics , quantum mechanics , gene , yeast
In this study, we hypothesize that ligand binding determines the binding partners and subcellular localization of nuclear receptors involved in atherosclerosis and diabetes. Specifically, we are interested in the effects of synthetic ligands on the peroxisome proliferator‐activated receptor alpha (PPARα), the liver X receptor alpha (LXRα) and the retinoid X receptor alpha (RXRα) and corresponding effects on heterodimer formation. Binding assays using a photon counting spectrofluorometer indicate promiscuity of protein‐ligand interactions where ligand binding is not solely exclusive. Additionally, Forster Resonance Energy Transfer (FRET) assays indicate that synthetic ligands affect the protein binding capability of the novel heterodimer PPARα:LXRα. Changes in secondary structure are anticipated, which would argue certain synthetic compounds can induce conformational changes that alter heterodimer formation. Bimolecular Fluorescence Complementation (BiFC) allows us to view protein‐protein interactions in vivo using fluorescence microscopy. We show that synthetic ligands influence subcellular localization of these nuclear receptors in stably transfected living cells. Due to the complex regulation of these nuclear receptors, it becomes necessary to study synthetic ligands with each receptor, individually and in heterodimeric form. Our findings will contribute to further the understanding of the clinical implications of synthetic ligands on disease states such as atherosclerosis and diabetes. Support or Funding Information This research is supported by USPHS NIH grant DK77573 and the Emerging Science Seed Grant provided by the Boonshoft School of Medicine and Wright State University.