Subject(s)
Fluorescence Resonance Energy Transfer/methods , Nuclear Proteins/metabolism , Receptors, Cytoplasmic and Nuclear/metabolism , Trans-Activators/metabolism , Transcription Factors/metabolism , Antibodies/metabolism , CREB-Binding Protein , Histone Acetyltransferases , Ligands , Nuclear Proteins/chemistry , Nuclear Proteins/genetics , Nuclear Receptor Coactivator 1 , Peptides/chemistry , Peptides/metabolism , Proto-Oncogene Proteins c-myb/immunology , Receptors, Cytoplasmic and Nuclear/chemistry , Recombinant Proteins/chemistry , Recombinant Proteins/metabolism , Trans-Activators/genetics , Transcription Factors/chemistry , Transcription Factors/geneticsABSTRACT
The mechanism by which ligands of nuclear receptors show differential effects on gene transcription is not fully understood, but is believed to result in part from the preferential recruitment and/or displacement of coactivators and corepressors. We have explored the interaction of several known ligands and the nuclear receptor (peroxisome proliferator activated receptor alpha, PPARalpha) using scintillation proximity assay (SPA) and the interaction of LXXLL containing peptides derived from three coactivators (SRC-1, CBP and PGC-1) with PPARalpha in the presence of PPARalpha agonist ligands using fluorescence resonance energy transfer (FRET). The EC(50)s of the individual ligands for recruitment showed the same rank order regardless of the coactivator peptide used, with GW2331Subject(s)
Receptors, Cytoplasmic and Nuclear/metabolism
, Transcription Factors/metabolism
, Cell Line
, Cell Nucleus/metabolism
, DNA, Complementary/metabolism
, Energy Transfer
, Escherichia coli/metabolism
, Histone Acetyltransferases
, Humans
, Kinetics
, Ligands
, Nuclear Receptor Coactivator 1
, Peptides/chemistry
, Peptides/metabolism
, Plasmids/metabolism
, Protein Binding
, Proto-Oncogene Proteins c-myc/metabolism
, Spectrophotometry
, Transfection
