ScholarWorks@숙명여자대학교

초록

Peroxisome proliferator-activated receptors (PPARs) are transcription factor which directly modulate gene expression by binding to specific agonists. It has been reported that PPAR alpha controls lipid metabolism, inflammation, and atherosclerosis. PPAR alpha activation by PPAR alpha agonist can ultimately reduce the progression of atherosclerosis and decrease the incidence of coronary heart disease. In this study, we optimized enzyme-linked immunosorbent assay (ELISA) systems in order to screen putative PPAR alpha agonists. These methods are based on the activation mechanism of PPAR alpha where the ligand binding to PPAR alpha induces the interaction of the receptor with transcriptional co-activators. Among co-activators such as SRC-1, TIF-2, and p300, although ligand-unbound PPAR alpha had more strong binding with p300 at a lower concentrations of PPAR alpha, ligand-bound PPARa had more specific and strong binding with SRC-1. We optimized and developed a novel and useful ELISA system to screen PPAR alpha agonists. Wyl 4,643 and linoleic acid, the wellknown PPAR alpha ligands, increased the binding between PPAR alpha and co-activators in a ligand dose-dependent manner. In this ELISA method to screen PPAR alpha ligands, the use of specific anti-PPAR alpha N-terminus antibody, full-length recombinant protein of human PPAR alpha but not ligand-binding domain (LBD) of human PPAR alpha, and his-tagged PPAR alpha recombinant proteins but not GST-fused PPAR alpha recombinant proteins is the critical factors. Development of this screening system may be useful in the discovery of PPAR alpha ligands from various candidates such as chemical library and phytochemicals.

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