A simple method to screen ligands of peroxisome proliferator-activated receptor delta.
Peroxisome proliferator-activated receptors (PPARs) are transcription factors that belong to the nuclear receptor superfamily directly modulating gene expression by binding to specific ligands. Recently, it has been reported that PPARdelta ligands play an essential role in improvement of metabolic disorders and skin disorders. We introduce an enzyme-linked immunosorbent assay (ELISA) to screen new PPARdelta ligands. This method is based on the activation mechanism of PPARdelta where the ligand binding to PPARdelta induces the interaction of the receptor with transcriptional co-activators. We optimized a simple ELISA method for screening PPARdelta ligands. Among co-activators such as SRC-1, TIF-2, and p300, PPARdelta had more strong binding with SRC-1 in an ELISA system. GW501516 and linoleic acid, the well-known ligands of PPARdelta, increased the binding between PPARdelta and co-activators in a ligand dose-dependent manner. The recruitment of co-activator SRC-1 was also more effective than those of TIF-2 and p300. We optimized and developed a novel and useful ELISA system for the mass screening of PPARdelta ligands. This screening system may be useful in the development of drugs for metabolic disorders and skin disorders.
Cho
Min-Chul
MC
Department of Bioscience & Biotechnology, Konkuk University, Seoul 143-701, Republic of Korea.
Yoon
Hyo-Eun
HE
Kang
Jeong-Woo
JW
Park
Se-Won
SW
Yang
Young
Y
Hong
Jin-Tae
JT
Song
Eun-Young
EY
Paik
Sang-Gi
SG
Kim
Soo-Hyun
SH
Yoon
Do-Young
DY
D003001:Cloning, Molecular; D004353:Drug Evaluation, Preclinical / Q000379:methods*; D050881:E1A-Associated p300 Protein / Q000737:chemistry / Q000378:metabolism; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods*; D004926:Escherichia coli / Q000378:metabolism; D051548:Histone Acetyltransferases / Q000737:chemistry / Q000378:metabolism; D008024:Ligands; D056920:Nuclear Receptor Coactivator 1; D047494:PPAR delta / Q000378:metabolism*; D011485:Protein Binding; D011965:Receptors, Glucocorticoid / Q000737:chemistry / Q000378:metabolism; D011994:Recombinant Proteins; D014157:Transcription Factors / Q000737:chemistry / Q000378:metabolism
D008024:Ligands; D047494:PPAR delta; D011965:Receptors, Glucocorticoid; D011994:Recombinant Proteins; D014157:Transcription Factors; D050881:E1A-Associated p300 Protein; C494360:EP300 protein, human; D051548:Histone Acetyltransferases; C538859:NCOA1 protein, human; D056920:Nuclear Receptor Coactivator 1