Laura Carpanedo, Luca M Wende, Bjarne Goebel, Ann-Kathrin Häfner, Michel André Chromik, Nadja Kampschulte, Dieter Steinhilber, Nils Helge Schebb
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Indirect quantification of esterified oxylipins in lipid fractions showed that >97% of each supplemented 15-LOX-2 product is esterified and that <25% are bound to neutral lipids, whereas >75% are bound to distinct glycero-PL classes, depending on the hydroxy-PUFA. 15-HETE and 15-HEPE were found in phosphatidylinositol (PI)/phosphatidylserine, whereas 17-HDHA was in phosphatidylethanolamine (PE) and 13-HODE in phosphatidylcholine (PC). The same pattern was found for oxylipins endogenously formed by overexpression of 15-LOX-2. A new targeted method for the analysis of oxidized glycero-PLs enabled to pinpoint the specific molecular species of the oxylipins. 15-HETE (20:4;15OH) and 15-HEPE (20:5;15OH) are dominantly found as PI 18:0/20:4;15OH (70%) and PI 18:0/20:5;15OH (80%), respectively. This preferential incorporation of 20:4;15OH and 20:5;15OH into PI may be biologically relevant for PI signaling pathways. In contrast, >50% of 17-HDHA (22:6;17OH) was found in PE P-16:0/22:6;17OH, PE P-18:0/22:6;17OH, and PE P-18:1/22:6;17OH. At least 40% of 13-HODE (18:2;13OH) was incorporated into PC 16:0/18:2;13OH, and relevant amounts were found in PI 18:0/18;13OH, PC 18:1/18;13OH, and PC-O (ether PC) 16:0/18;13OH. These results indicate that hydroxy-PUFAs are bound to glycero-PLs in a specific manner. The distinct incorporation of 15-LOX-2 products from different PUFAs into glycero-PLs might contribute to the biological effect of these oxylipins and their precursor FAs.</p>","PeriodicalId":16209,"journal":{"name":"Journal of Lipid Research","volume":" ","pages":"100841"},"PeriodicalIF":5.0000,"publicationDate":"2025-06-14","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12274753/pdf/","citationCount":"0","resultStr":"{\"title\":\"Substrate-dependent incorporation of 15-lipoxygenase products in glycerophospholipids: 15-HETE and 15-HEPE in PI, 17-HDHA in plasmalogen PE, and 13-HODE in PC.\",\"authors\":\"Laura Carpanedo, Luca M Wende, Bjarne Goebel, Ann-Kathrin Häfner, Michel André Chromik, Nadja Kampschulte, Dieter Steinhilber, Nils Helge Schebb\",\"doi\":\"10.1016/j.jlr.2025.100841\",\"DOIUrl\":null,\"url\":null,\"abstract\":\"<p><p>Several oxylipins including hydroxy-PUFAs act as lipid mediators. In biological samples, the major part occurs esterified in glycero-phospholipids (PLs) or other lipids. In this work, the incorporation into glycero-PLs of 15-hydroxyeicosatetraenoic acid (15(S)-HETE), 15(S)-hydroxyeicosapentaenoic acid (15(S)-HEPE), 17(S)-hydroxydocosahexaenoic acid (17(S)-HDHA), and 13-(S)-hydroxyoctadecadienoic acid (13(S)-HODE) was investigated in oxylipin-supplemented human embryonic kidney 293T cells and cells overexpressing 15-lipoxgenase-2 (15-LOX-2, ALOX15B). Indirect quantification of esterified oxylipins in lipid fractions showed that >97% of each supplemented 15-LOX-2 product is esterified and that <25% are bound to neutral lipids, whereas >75% are bound to distinct glycero-PL classes, depending on the hydroxy-PUFA. 15-HETE and 15-HEPE were found in phosphatidylinositol (PI)/phosphatidylserine, whereas 17-HDHA was in phosphatidylethanolamine (PE) and 13-HODE in phosphatidylcholine (PC). The same pattern was found for oxylipins endogenously formed by overexpression of 15-LOX-2. A new targeted method for the analysis of oxidized glycero-PLs enabled to pinpoint the specific molecular species of the oxylipins. 15-HETE (20:4;15OH) and 15-HEPE (20:5;15OH) are dominantly found as PI 18:0/20:4;15OH (70%) and PI 18:0/20:5;15OH (80%), respectively. This preferential incorporation of 20:4;15OH and 20:5;15OH into PI may be biologically relevant for PI signaling pathways. In contrast, >50% of 17-HDHA (22:6;17OH) was found in PE P-16:0/22:6;17OH, PE P-18:0/22:6;17OH, and PE P-18:1/22:6;17OH. At least 40% of 13-HODE (18:2;13OH) was incorporated into PC 16:0/18:2;13OH, and relevant amounts were found in PI 18:0/18;13OH, PC 18:1/18;13OH, and PC-O (ether PC) 16:0/18;13OH. These results indicate that hydroxy-PUFAs are bound to glycero-PLs in a specific manner. The distinct incorporation of 15-LOX-2 products from different PUFAs into glycero-PLs might contribute to the biological effect of these oxylipins and their precursor FAs.</p>\",\"PeriodicalId\":16209,\"journal\":{\"name\":\"Journal of Lipid Research\",\"volume\":\" \",\"pages\":\"100841\"},\"PeriodicalIF\":5.0000,\"publicationDate\":\"2025-06-14\",\"publicationTypes\":\"Journal Article\",\"fieldsOfStudy\":null,\"isOpenAccess\":false,\"openAccessPdf\":\"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12274753/pdf/\",\"citationCount\":\"0\",\"resultStr\":null,\"platform\":\"Semanticscholar\",\"paperid\":null,\"PeriodicalName\":\"Journal of Lipid Research\",\"FirstCategoryId\":\"99\",\"ListUrlMain\":\"https://doi.org/10.1016/j.jlr.2025.100841\",\"RegionNum\":2,\"RegionCategory\":\"医学\",\"ArticlePicture\":[],\"TitleCN\":null,\"AbstractTextCN\":null,\"PMCID\":null,\"EPubDate\":\"\",\"PubModel\":\"\",\"JCR\":\"Q1\",\"JCRName\":\"BIOCHEMISTRY & MOLECULAR BIOLOGY\",\"Score\":null,\"Total\":0}","platform":"Semanticscholar","paperid":null,"PeriodicalName":"Journal of Lipid Research","FirstCategoryId":"99","ListUrlMain":"https://doi.org/10.1016/j.jlr.2025.100841","RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"Q1","JCRName":"BIOCHEMISTRY & MOLECULAR BIOLOGY","Score":null,"Total":0}
Substrate-dependent incorporation of 15-lipoxygenase products in glycerophospholipids: 15-HETE and 15-HEPE in PI, 17-HDHA in plasmalogen PE, and 13-HODE in PC.
Several oxylipins including hydroxy-PUFAs act as lipid mediators. In biological samples, the major part occurs esterified in glycero-phospholipids (PLs) or other lipids. In this work, the incorporation into glycero-PLs of 15-hydroxyeicosatetraenoic acid (15(S)-HETE), 15(S)-hydroxyeicosapentaenoic acid (15(S)-HEPE), 17(S)-hydroxydocosahexaenoic acid (17(S)-HDHA), and 13-(S)-hydroxyoctadecadienoic acid (13(S)-HODE) was investigated in oxylipin-supplemented human embryonic kidney 293T cells and cells overexpressing 15-lipoxgenase-2 (15-LOX-2, ALOX15B). Indirect quantification of esterified oxylipins in lipid fractions showed that >97% of each supplemented 15-LOX-2 product is esterified and that <25% are bound to neutral lipids, whereas >75% are bound to distinct glycero-PL classes, depending on the hydroxy-PUFA. 15-HETE and 15-HEPE were found in phosphatidylinositol (PI)/phosphatidylserine, whereas 17-HDHA was in phosphatidylethanolamine (PE) and 13-HODE in phosphatidylcholine (PC). The same pattern was found for oxylipins endogenously formed by overexpression of 15-LOX-2. A new targeted method for the analysis of oxidized glycero-PLs enabled to pinpoint the specific molecular species of the oxylipins. 15-HETE (20:4;15OH) and 15-HEPE (20:5;15OH) are dominantly found as PI 18:0/20:4;15OH (70%) and PI 18:0/20:5;15OH (80%), respectively. This preferential incorporation of 20:4;15OH and 20:5;15OH into PI may be biologically relevant for PI signaling pathways. In contrast, >50% of 17-HDHA (22:6;17OH) was found in PE P-16:0/22:6;17OH, PE P-18:0/22:6;17OH, and PE P-18:1/22:6;17OH. At least 40% of 13-HODE (18:2;13OH) was incorporated into PC 16:0/18:2;13OH, and relevant amounts were found in PI 18:0/18;13OH, PC 18:1/18;13OH, and PC-O (ether PC) 16:0/18;13OH. These results indicate that hydroxy-PUFAs are bound to glycero-PLs in a specific manner. The distinct incorporation of 15-LOX-2 products from different PUFAs into glycero-PLs might contribute to the biological effect of these oxylipins and their precursor FAs.
期刊介绍：
The Journal of Lipid Research (JLR) publishes original articles and reviews in the broadly defined area of biological lipids. We encourage the submission of manuscripts relating to lipids, including those addressing problems in biochemistry, molecular biology, structural biology, cell biology, genetics, molecular medicine, clinical medicine and metabolism. Major criteria for acceptance of articles are new insights into mechanisms of lipid function and metabolism and/or genes regulating lipid metabolism along with sound primary experimental data. Interpretation of the data is the authors’ responsibility, and speculation should be labeled as such. Manuscripts that provide new ways of purifying, identifying and quantifying lipids are invited for the Methods section of the Journal. JLR encourages contributions from investigators in all countries, but articles must be submitted in clear and concise English.
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