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タイトル: | Development of a Novel PPARγ Ligand Screening System Using Pinpoint Fluorescence-Probed Protein |
著者: | NAGAI, Hiroyuki EBISU, Shogo ABE, Ryoji GOTO, Tsuyoshi https://orcid.org/0000-0003-1283-147X (unconfirmed) TAKAHASHI, Nobuyuki HOSAKA, Takahiro KAWADA, Teruo |
著者名の別形: | 河田, 照雄 |
キーワード: | peroxisome-proliferator-activated receptor-γ (PPARγ) co-activator high-throughput ligand screening system |
発行日: | Feb-2011 |
出版者: | Japan Society for Bioscience, Biotechnology, and Agrochemistry |
誌名: | Bioscience, Biotechnology, and Biochemistry |
巻: | 75 |
号: | 2 |
開始ページ: | 337 |
終了ページ: | 341 |
抄録: | The activation of peroxisome-proliferator-activated receptor-γ (PPARγ), which plays a central role in adipocyte differentiation, depends on ligand-dependent co-activator recruitment. In this study, we developed a novel method of PPARγ ligand screening by measuring the increase in fluorescent polarization accompanied by the interaction of a fluorescent co-activator and PPARγ. Sterol receptor co-activator-1 (SRC-1), a major PPARγ co-activator, was probed by fluorescent TAMRA by the Amber codon fluorescence probe method. Polarization was increased by adding PPARγ ligands to a solution containing labeled SRC-1 (designated TAMRA-SRC-S) and PPARγ. The disassociation constants (Kd) of the PPARγ synthesized ligands, pioglitazone (221 nM), troglitazone (83.0 nM), and 15-deoxy-Δ12, 14-prostaglandin J2 (15d-ΔPGJ2) (156 nM), were determined by this method. Farnesol (2.89 μM) and bixin (21.1 μM), which we have reported to be PPARγ ligands, increased the fluorescent polarization. Their Kd values were in agreement with the ED50 values obtained in the luciferase assay. The results indicate that the method is valuable for screening natural PPARγ ligands. |
著作権等: | (c) 2011 by Japan Society for Bioscience, Biotechnology, and Agrochemistry |
URI: | http://hdl.handle.net/2433/156398 |
DOI(出版社版): | 10.1271/bbb.100810 |
PubMed ID: | 21307572 |
関連リンク: | https://www.jstage.jst.go.jp/article/bbb/75/2/75_100810/_article |
出現コレクション: | 学術雑誌掲載論文等 |
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