Enantioselective activation of the peroxisome proliferator-activated receptor.
A cell-based transactivation assay was established using the mouse full-length peroxisome proliferator-activated receptor (PPAR) cDNA sequence and the positive peroxisome proliferator-responsive regulatory element (-578 to -553) of the rat acyl-CoA oxidase gene promoter. Activation of the reporter plasmid was dependent on co-transfection of the full-length PPAR cDNA, and the response was greatly stimulated, up to 100-fold, by peroxisome proliferators such as Wy-14,643 ([4-chloro-6-(2,3-xylidino)-2-pyrimidinylthio] acetic acid), nafenopin (2-methyl-2[p-(1,2,3,4-tetrahydro-1-naphthyl)phenoxy]-propionic acid), and clofibric acid (2-([p]-chlorophenoxy)-2-methylpropionic acid). Activation of the reporter plasmid promoter by the full-length PPAR cDNA also occurred at peroxisomal proliferator concentrations 40 times lower than that required for similar stimulation by a glucocorticoid-PPAR chimeric receptor. By using the stereoisomers of MK-571 ((+-)-3-(((3-(2-(7-chloro-2-quinolinyl)ethenyl)-phenyl)((3- (dimethylamino)-3-oxopropyl)-thio)methyl)-thio)propanoic acid), a potent leukotriene D4 receptor antagonist, we could show enantioselective activation of PPAR. The use of this compound in mice results in peroxisome proliferation; however, nearly all of the peroxisome proliferating activity can be attributed to the S enantiomer. Our results show a similar enantiomeric discrimination in PPAR activation of the reporter plasmid promoter, where again most of the activity can be attributed to the S enantiomer. The equivalent activities of these stereoisomers both in vivo and in the PPAR transactivation assay strongly implicate PPAR as a major component of the peroxisome proliferating mechanism in rodents.
Boie
Y
Y
Department of Molecular Biology, Merck Frosst Centre for Therapeutic Research, Pointe Claire-Dorval, Quebec, Canada.
Adam
M
M
Rushmore
T H
TH
Kennedy
B P
BP
D042962:Acyl-CoA Oxidase; D000818:Animals; D001483:Base Sequence; D002478:Cells, Cultured; D002995:Clofibric Acid / Q000494:pharmacology; D004247:DNA; D008830:Microbodies / Q000187:drug effects; D008969:Molecular Sequence Data; D009255:Nafenopin / Q000494:pharmacology; D010088:Oxidoreductases / Q000235:genetics*; D011422:Propionates / Q000737:chemistry / Q000494:pharmacology; D011743:Pyrimidines / Q000494:pharmacology; D011804:Quinolines / Q000737:chemistry / Q000494:pharmacology; D051381:Rats; D011956:Receptors, Cell Surface / Q000187:drug effects / Q000235:genetics / Q000378:metabolism*; D018160:Receptors, Cytoplasmic and Nuclear; D012045:Regulatory Sequences, Nucleic Acid; D013237:Stereoisomerism; D014157:Transcription Factors / Q000378:metabolism*; D015533:Transcriptional Activation
D011422:Propionates; D011743:Pyrimidines; D011804:Quinolines; D011956:Receptors, Cell Surface; D018160:Receptors, Cytoplasmic and Nuclear; D014157:Transcription Factors; D009255:Nafenopin; D002995:Clofibric Acid; C059141:verlukast; C006253:pirinixic acid; D004247:DNA; D010088:Oxidoreductases; D042962:Acyl-CoA Oxidase