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 ac...

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Main Authors: Laura Carpanedo, Luca M. Wende, Bjarne Goebel, Ann-Kathrin Häfner, Michel André Chromik, Nadja Kampschulte, Dieter Steinhilber, Nils Helge Schebb
Format: Article
Language:English
Published: Elsevier 2025-07-01
Series:Journal of Lipid Research
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Online Access:http://www.sciencedirect.com/science/article/pii/S0022227525001014
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author Laura Carpanedo
Luca M. Wende
Bjarne Goebel
Ann-Kathrin Häfner
Michel André Chromik
Nadja Kampschulte
Dieter Steinhilber
Nils Helge Schebb
author_facet Laura Carpanedo
Luca M. Wende
Bjarne Goebel
Ann-Kathrin Häfner
Michel André Chromik
Nadja Kampschulte
Dieter Steinhilber
Nils Helge Schebb
author_sort Laura Carpanedo
collection DOAJ
description 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.
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spelling doaj-art-80049a761f614f8389cfb9347efaba302025-08-20T03:14:34ZengElsevierJournal of Lipid Research0022-22752025-07-0166710084110.1016/j.jlr.2025.100841Substrate-dependent incorporation of 15-lipoxygenase products in glycerophospholipids: 15-HETE and 15-HEPE in PI, 17-HDHA in plasmalogen PE, and 13-HODE in PCLaura Carpanedo0Luca M. Wende1Bjarne Goebel2Ann-Kathrin Häfner3Michel André Chromik4Nadja Kampschulte5Dieter Steinhilber6Nils Helge Schebb7Chair of Food Chemistry, Faculty of Mathematics and Natural Sciences, University of Wuppertal, Wuppertal, GermanyChair of Food Chemistry, Faculty of Mathematics and Natural Sciences, University of Wuppertal, Wuppertal, GermanyInstitute of Pharmaceutical Chemistry, Goethe University Frankfurt, Frankfurt, GermanyInstitute of Pharmaceutical Chemistry, Goethe University Frankfurt, Frankfurt, GermanyChair of Food Chemistry, Faculty of Mathematics and Natural Sciences, University of Wuppertal, Wuppertal, GermanyChair of Food Chemistry, Faculty of Mathematics and Natural Sciences, University of Wuppertal, Wuppertal, GermanyInstitute of Pharmaceutical Chemistry, Goethe University Frankfurt, Frankfurt, GermanyChair of Food Chemistry, Faculty of Mathematics and Natural Sciences, University of Wuppertal, Wuppertal, Germany; For correspondence: Nils Helge SchebbSeveral 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.http://www.sciencedirect.com/science/article/pii/S0022227525001014oxidized lipidslipidomicscell signalinginflammationarachidonic acidoxylipin incorporation
spellingShingle Laura Carpanedo
Luca M. Wende
Bjarne Goebel
Ann-Kathrin Häfner
Michel André Chromik
Nadja Kampschulte
Dieter Steinhilber
Nils Helge Schebb
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
Journal of Lipid Research
oxidized lipids
lipidomics
cell signaling
inflammation
arachidonic acid
oxylipin incorporation
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
title_full 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
title_fullStr 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
title_full_unstemmed 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
title_short 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
title_sort 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
topic oxidized lipids
lipidomics
cell signaling
inflammation
arachidonic acid
oxylipin incorporation
url http://www.sciencedirect.com/science/article/pii/S0022227525001014
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