title: Bidirectional fluorescence properties of pyrene-based peroxisome proliferator-activated receptor (PPAR) α/δ dual agonist.
abstract: Based on X-ray crystallographic analysis of a peroxisome proliferator-activated receptor (PPAR) α/δ dual agonist complexed with human PPARs ligand binding domain (LBD), we previously reported the design and synthesis of a pyrene-based fluorescent PPARα/δ co-agonist 2. Here, we found that the fluorescence intensity of 2 increased upon binding to hPPARα-LBD, in a manner dependent upon the concentration of the LBD. But, surprisingly, the fluorescence intensity of 2 decreased concentration-dependently upon binding to hPPRδ-LBD. Site-directed mutagenesis of the two hPPAR subtypes clearly indicated that Trp264 of hPPARδ-LBD, located between H2' helix and H3 helix (omega loop), is critical for the concentration-dependent decrease in fluorescence intensity, which is suggested to be due to fluorescence resonance energy transfer (FRET) from the pyrene moiety of bound 2 to the nearby side-chain indole moiety of Trp264 in the hPPARδ-LBD.
authors.affiliation: Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, Okayama University, 1-1-1, Tsushima-Naka, Kita-ku, Okayama 700-8530, Japan.
authors.forename: Shintaro
authors.initials: S
authors.lastname: Ban
authors.forename: Takuji
authors.initials: T
authors.lastname: Oyama
authors.forename: Jun-ichi
authors.initials: J
authors.lastname: Kasuga
authors.forename: Kenji
authors.initials: K
authors.lastname: Ohgane
authors.forename: Yoshino
authors.initials: Y
authors.lastname: Nishio
authors.forename: Kosuke
authors.initials: K
authors.lastname: Morikawa
authors.forename: Yuichi
authors.initials: Y
authors.lastname: Hashimoto
authors.forename: Hiroyuki
authors.initials: H
authors.lastname: Miyachi
mesh_terms: D000595:Amino Acid Sequence; D001665:Binding Sites; D015195:Drug Design; D005453:Fluorescence; D031541:Fluorescence Resonance Energy Transfer; D006801:Humans; D008024:Ligands; D008958:Models, Molecular; D008969:Molecular Sequence Data; D016297:Mutagenesis, Site-Directed; D047493:PPAR alpha / Q000819:agonists* / Q000235:genetics / Q000378:metabolism; D047494:PPAR delta / Q000819:agonists* / Q000235:genetics / Q000378:metabolism; D010666:Phenylpropionates / Q000737:chemistry* / Q000378:metabolism* / Q000494:pharmacology; D011487:Protein Conformation; D011721:Pyrenes / Q000737:chemistry* / Q000378:metabolism* / Q000494:pharmacology; D014364:Tryptophan
chemical_list: C575346:2-(4-methoxy-3-((pyrene-1-carboxamido)methyl)benzyl)butanoic acid; D008024:Ligands; D047493:PPAR alpha; D047494:PPAR delta; D010666:Phenylpropionates; D011721:Pyrenes; D014364:Tryptophan