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KMID : 1094720170220010100
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Biotechnology and Bioprocess Engineering
2017 Volume.22 No. 1 p.100 ~ p.106
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Quantitative targeted metabolomics for 15d-deoxy-Δ12, 14-PGJ2 (15d-PGJ2) by MALDI-MS
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Kim Kyoung-Jin
Park Han-Gyu
Hwang Cheol-Hwan
Ann Da-Hee
Song Won-Suk
Choi Kwon-Young
Yang Yung-Hun
Park Sang-Youn
Kim Yun-Gon
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Abstract
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Prostaglandins (PGs) are lipid mediators that may play important roles in cancer, immunomodulation, and neurodegeneration. So, the quantitative analysis of PGs will therefore be important in order to understand the natural history of a range of diseases and may be used as a tool in the development of new biotherapeutics. However, such an analysis is problematic because of the small quantities of PGs present in the body. Here, we developed a matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-MS)-based analytical method for rapid and easy quantification of the ketone-containing PGs (15d-PGJ2 etc.) as a targeted metabolomics platform. The chemical derivatization with Girard’s reagent P (GP) provided a good linearity (R2 > 0.99) between peak area and quantity of 15d-PGJ2 and highly improved sensitivity (limit of quantitation, LOQ: 1.25 pmol on a spot in MALDI plate). Finally, we utilized this method to directly characterize the interaction between peroxisome proliferatoractivated receptor gamma-ligand binding domain (PPARγ-LBD) and 15d-PGJ2. The 15d-PGJ2 was enriched by PPARγ-LBD and also the binding level of the ligand was dropped considerably by the treatment of PPARγ agonist such as rosiglitazone (about 0.61-fold reduction). Taken together, our MALDI-MS-based targeted metabolomics method for ketone-containing PGs may be applicable to elucidate the protein-metabolite interactions and to identify natural ligands for drug candidates.
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KEYWORD
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Ketone-containing prostaglandins, 15d-PGJ2, MALDI-MS, Girard’s reagent P, PPARγ-LBD
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