TY - JOUR
T1 - Bidirectional fluorescence properties of pyrene-based peroxisome proliferator-activated receptor (PPAR) α/δ dual agonist
AU - Ban, Shintaro
AU - Oyama, Takuji
AU - Kasuga, Jun Ichi
AU - Ohgane, Kenji
AU - Nishio, Yoshino
AU - Morikawa, Kosuke
AU - Hashimoto, Yuichi
AU - Miyachi, Hiroyuki
N1 - Funding Information:
This work was supported in part by the Targeted Proteins Research Program of the Japan Science and Technology Corporation (JST), the Uehara Memorial Foundation, the Tokyo Biochemical Research Foundation (TBRF) and the Okayama Foundation for Science and Technology (OFST).
PY - 2012/6/1
Y1 - 2012/6/1
N2 - 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.
AB - 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.
KW - Fluorescent PPARα/δ co-agonist
KW - PPARα
KW - PPARδ
KW - Peroxisome proliferator-activated receptor
KW - Site-directed mutagenesis
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U2 - 10.1016/j.bmc.2012.04.015
DO - 10.1016/j.bmc.2012.04.015
M3 - Article
C2 - 22551628
AN - SCOPUS:84861188794
SN - 0968-0896
VL - 20
SP - 3460
EP - 3464
JO - Bioorganic and Medicinal Chemistry
JF - Bioorganic and Medicinal Chemistry
IS - 11
ER -