Acyl-CoA Esters Antagonize the Effects of Ligands on Peroxisome Proliferator-activated Receptor α Conformation, DNA Binding, and Interaction with Co-factors

Morten Elholm, Inge Dam, Claus Jørgensen, Anne M. Krogsdam, Dorte Holst, Irina Kratchmarova, Martin Göttlicher, Jan Åke Gustafsson, Rolf Berge, Torgeir Flatmark, Jens Knudsen, Susanne Mandrup, Karsten Kristiansen

Research output: Contribution to journalArticle

38 Scopus citations

Abstract

The peroxisome proliferator-activated receptor α (PPARα) is a ligand-activated transcription factor and a key regulator of lipid homeostasis. Numerous fatty acids and eicosanoids serve as ligands and activators for PPARα. Here we demonstrate that S-hexadecyl-CoA, a nonhydrolyzable palmitoyl-CoA analog, antagonizes the effects of agonists on PPARα conformation and function in vitro. In electrophoretic mobility shift assays, S-hexadecyl-CoA prevented agonist-induced binding of the PPARα-retinoid X receptor α heterodimer to the acyl-CoA oxidase peroxisome proliferator response element. PPARα bound specifically to immobilized palmitoyl-CoA and Wy14643, but not BRL49653, abolished binding. S-Hexadecyl-CoA increased in a dose-dependent and reversible manner the sensitivity of PPARα to chymotrypsin digestion, and the S-hexadecyl-CoA-induced sensitivity required a functional PPARα ligand-binding pocket. S-Hexadecyl-CoA prevented ligand-induced interaction between the co-activator SRC-1 and PPARα but increased recruitment of the nuclear receptor co-repressor NCoR. In cells, the concentration of free acyl-CoA esters is kept in the low nanomolar range due to the buffering effect of high affinity acyl-CoA-binding proteins, especially the acyl-CoA-binding protein. By using PPARα expressed in Sf21 cells for electrophoretic mobility shift assays, we demonstrate that S-hexadecyl-CoA was able to increase the mobility of the PPARα-containing heterodimer even in the presence of a molar excess of acyl-CoA-binding protein, mimicking the conditions found in vivo.

Original languageEnglish (US)
Pages (from-to)21410-21416
Number of pages7
JournalJournal of Biological Chemistry
Volume276
Issue number24
DOIs
StatePublished - Jun 15 2001

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology
  • Cell Biology

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