TY - JOUR
T1 - Identification of novel oxidized phospholipids that activate platelet-activating factor receptor using HPLC fractionation and comprehensive LC-MS/MS analysis
AU - Kozakura, Eisho
AU - Ueno, Ryoya
AU - Yamashita, Tomohiro
AU - Hashidate-Yoshida, Tomomi
AU - Shindou, Hideo
AU - Jutanom, Mirinthorn
AU - Morimoto, Kazushi
AU - Yamada, Ken ichi
N1 - Publisher Copyright:
© 2025
PY - 2025/6/8
Y1 - 2025/6/8
N2 - Platelet-activating factor receptor (PAFR) is involved in various physiological processes, including the immune system and inflammatory responses. In addition to PAF, several oxidized phospholipids have been shown to act as ligands for PAFR. We have previously developed a comprehensive analysis method for oxidized phospholipids, and in this study, we employed this method to test whether additional oxidized phospholipids can activate PAFR. From an oxidized phosphatidylcholine mixture, we identified that 1-palmitoyl-2-(4′-oxo-butanoyl)-sn-glycero-3-phosphocholine (POBPC) functions as a novel PAFR activator, using preparative HPLC and comprehensive LC-MS/MS analysis of fractionated oxidized phospholipids. Next, multiple assays confirmed that POBPC acts as a bona fide PAFR agonist. The H248W mutation of PAFR attenuated the response to POBPC. Finally, POBPC induced phosphorylation of extracellular signal-regulated kinase in mouse peritoneal macrophages, which endogenously express PAFR. Our findings provide valuable insight into the biological functions of oxidized phospholipids, advancing our understanding of their roles in cellular processes.
AB - Platelet-activating factor receptor (PAFR) is involved in various physiological processes, including the immune system and inflammatory responses. In addition to PAF, several oxidized phospholipids have been shown to act as ligands for PAFR. We have previously developed a comprehensive analysis method for oxidized phospholipids, and in this study, we employed this method to test whether additional oxidized phospholipids can activate PAFR. From an oxidized phosphatidylcholine mixture, we identified that 1-palmitoyl-2-(4′-oxo-butanoyl)-sn-glycero-3-phosphocholine (POBPC) functions as a novel PAFR activator, using preparative HPLC and comprehensive LC-MS/MS analysis of fractionated oxidized phospholipids. Next, multiple assays confirmed that POBPC acts as a bona fide PAFR agonist. The H248W mutation of PAFR attenuated the response to POBPC. Finally, POBPC induced phosphorylation of extracellular signal-regulated kinase in mouse peritoneal macrophages, which endogenously express PAFR. Our findings provide valuable insight into the biological functions of oxidized phospholipids, advancing our understanding of their roles in cellular processes.
KW - HPLC
KW - LC-MS/MS
KW - Oxidized phosphatidylcholine
KW - PAFR
KW - POBPC
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U2 - 10.1016/j.bbrc.2025.151858
DO - 10.1016/j.bbrc.2025.151858
M3 - Article
AN - SCOPUS:105003174194
SN - 0006-291X
VL - 765
JO - Biochemical and Biophysical Research Communications
JF - Biochemical and Biophysical Research Communications
M1 - 151858
ER -