A simple ELISA for screening ligands of peroxisome proliferator-activated receptor-gamma.
Peroxisome proliferator-activated receptors (PPARs) are orphan nuclear hormone receptors that are known to control the expression of genes that are involved in lipid homeostasis and energy balance. PPARs activate gene transcription in response to a variety of compounds, including hypolipidemic drugs. Most of these compounds have high affinity to the ligand-binding domain (LBD) of PPARs and cause a conformational change within PPARs. As a result, the receptor is converted to an activated mode that promotes the recruitment of co-activators such as the steroid receptor co-activator-1 (SRC-1). Based on the activation mechanism of PPARs (the ligand binding to PPAR gamma induces interactions of the receptor with transcriptional co-activators), we performed Western blot and ELISA. These showed that the indomethacin, a PPAR gamma ligand, increased the binding between PPAR gamma and SRC-1 in a ligand dose-dependent manner. These results suggested that the in vitro conformational change of PPAR gamma by ligands was also induced, and increased the levels of the ligand-dependent interaction with SRC-1. Collectively, we developed a novel and useful ELISA system for the mass screening of PPAR gamma ligands. This screening system (based on the interaction between PPAR gamma and SRC-1) may be a promising system in the development of drugs for metabolic disorders.
Cho
Min Chul
MC
Laboractory of Cellular Biology, Korea Research Institute of Bioscience and Biotechnology, Daejon 305-600, Korea.
Lee
Hae Sook
HS
Kim
Jae Hwa
JH
Choe
Yong Kyung
YK
Hong
Jin Tae
JT
Paik
Sang Gi
SG
Yoon
Do Young
DY
D000818:Animals; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods*; D051548:Histone Acetyltransferases; D006801:Humans; D007213:Indomethacin / Q000494:pharmacology; D008024:Ligands; D051379:Mice; D056920:Nuclear Receptor Coactivator 1; D011485:Protein Binding; D018160:Receptors, Cytoplasmic and Nuclear / Q000037:antagonists & inhibitors / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D011994:Recombinant Proteins / Q000302:isolation & purification / Q000378:metabolism; D014157:Transcription Factors / Q000037:antagonists & inhibitors / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*
D008024:Ligands; D018160:Receptors, Cytoplasmic and Nuclear; D011994:Recombinant Proteins; D014157:Transcription Factors; D051548:Histone Acetyltransferases; C538859:NCOA1 protein, human; C538861:Ncoa1 protein, mouse; D056920:Nuclear Receptor Coactivator 1; D007213:Indomethacin