Journal of Lipid Research最新文献

{"title":"Functional characterization of genetic variants affecting the intracellular domains of ATP-binding cassette transporter A1 (ABCA1).","authors":"Marianne Teigen, Åsa Schawlann Ølnes, Katrine Bjune, Martin Prøven Bogsrud, Thea Bismo Strøm","doi":"10.1016/j.jlr.2025.100854","DOIUrl":"https://doi.org/10.1016/j.jlr.2025.100854","url":null,"abstract":"<p><p>The ATP-binding cassette transporter A1 (ABCA1) effluxes cellular cholesterol and phospholipids to extracellular acceptors, mainly apolipoprotein A1, generating high density lipoprotein (HDL) particles. This is the first step in the anti-atherosclerotic process of transporting excess cholesterol from non-hepatic tissues to the liver. Loss-of-function variants in ABCA1 lead to reduced HDL cholesterol levels in plasma, thus possibly diminishing the atheroprotective effect of HDL. More than 250 missense variants have been reported in the ABCA1 gene, most of which remain to be functionally characterized. In this study we have characterized 74 variants affecting the intracellular domains of ABCA1 by assessing cholesterol efflux activity and cell surface localization of the protein, thereby shifting the pathogenicity classification of 10 variants from class 3 (uncertain significance) to class 4 (likely pathogenic) or class 2 (likely benign). Consequently, functional characterization contributes to a better understanding of the molecular basis of the pathogenicity of genetic variants in ABCA1, which could also clarify the mechanism of action of the protein.</p>","PeriodicalId":16209,"journal":{"name":"Journal of Lipid Research","volume":" ","pages":"100854"},"PeriodicalIF":5.0,"publicationDate":"2025-07-03","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144567526","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Daytime-restricted feeding induces lean MAFLD in high-fat diet-fed mice by upregulating CD36-mediated lipid accumulation.","authors":"Zhenyu Wang, Mingyang Zhang, Miao Chen, Shuning Fu, Yang Zhang, Mengyue Chen, Xiong Z Ruan, Yaxi Chen","doi":"10.1016/j.jlr.2025.100853","DOIUrl":"10.1016/j.jlr.2025.100853","url":null,"abstract":"<p><p>Time-restricted feeding (TRF) may aid in weight loss and improve metabolic health; however, its long-term effects and applicability to all individuals remain unclear. This study investigated the impact of different dietary patterns on hepatic metabolism by subjecting mice to either a normal chow diet or a high-fat diet, allowing for ad libitum feeding, daytime restrictive feeding (DRF), or nighttime restrictive feeding (NRF). Using metabolic cages to assess energy intake, we found that the fuel utilization rhythms of DRF mice were disrupted compared to ad libitum-fed mice. Mice on normal chow DRF exhibited only dyslipidemia, while those on high-fat DRF developed lean metabolic dysfunction-associated fatty liver disease (MAFLD), characterized by more pronounced dyslipidemia, weight loss, and hepatic lipid accumulation. RNA seq revealed that CD36 plays a crucial role in the development of lean MAFLD induced by high-fat DRF by inhibiting AMPK phosphorylation, disrupting the balance between lipogenesis and oxidation. Mechanistic validation was performed in CD36 liver-specific knockout mice and Liposomal nanoparticle injection models. These findings provide new insights into the potential mechanisms linking feeding patterns to lean MAFLD. Additionally, CD36 emerges as a potential therapeutic target for high-fat-induced lean MAFLD. Clarifying the relationship between DRF and lean MAFLD may inform guidelines for specific populations, such as individuals practicing intermittent fasting or those working night shifts, while also suggesting potential therapeutic strategies for clinical management.</p>","PeriodicalId":16209,"journal":{"name":"Journal of Lipid Research","volume":" ","pages":"100853"},"PeriodicalIF":5.0,"publicationDate":"2025-06-23","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144497337","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Modeling the postprandial CETP-mediated lipid redistribution between chylomicrons, LDL and HDL.","authors":"Martin Jansen, Christine Contini, Michael M Hoffmann, Gerhard Puetz","doi":"10.1016/j.jlr.2025.100847","DOIUrl":"10.1016/j.jlr.2025.100847","url":null,"abstract":"<p><p>Impaired triglyceride (TG) metabolism is associated with metabolic diseases. Non-steady state dynamics make studying postprandial lipid metabolism challenging. We already introduced a mathematical model to estimate cholesteryl ester transfer protein (CETP)-mediated TG net flux in the fasting state. Here, we expand this model to chylomicrons (CMs) and the dynamics of postprandial lipemia. Blood samples of normolipidemic, hypertriglyceridemic, and hyperchylomicronemic volunteers were drawn at fasting and postprandial state. We separated lipoprotein classes via classical sequential ultracentrifugation. To address CMs, we developed a novel method based on Airfuge® ultracentrifugation. We studied postprandial changes of lipoproteins and their components. CETP-mediated TG redistribution was modeled based on the surface and composition data of respective lipoprotein fractions and validated by corresponding measured values. Our model estimated CETP-mediated TG flux in the fasting and postprandial state with high accuracy. Even in the postprandial condition, TG net flux to LDL/HDL is dominated by VLDL. Separating CM from VLDL and modeling both fractions instead of just using the combined CM + VLDL fraction did only improve the model's accuracy slightly (by less than 7%). The proportion of ApoC3 redistributed from HDL to VLDL in postprandial lipemia is highly correlated with the change of ApoA1 in HDL2b. Our basic model is able to estimate TG redistribution via CETP among lipoproteins in postprandial lipemia of healthy and hypertriglyceridemic subjects. An additional separation of VLDL and CM is not strictly necessary to model postprandial TG flux. Our model makes postprandial lipoprotein metabolism more tangible and may help to study lipoprotein-associated pathologies.</p>","PeriodicalId":16209,"journal":{"name":"Journal of Lipid Research","volume":" ","pages":"100847"},"PeriodicalIF":5.0,"publicationDate":"2025-06-23","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144497348","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Nanodisc single-molecule pulldown to study lipid-protein interactions.","authors":"Adriana Reyes-Ordoñez, Shweta Shree, Nilmani Singh, Stephen G Sligar, Jie Chen","doi":"10.1016/j.jlr.2025.100846","DOIUrl":"10.1016/j.jlr.2025.100846","url":null,"abstract":"<p><p>Beyond serving structural roles in the cell membrane, many phospholipids, including phosphatidylinositol phosphates (PIPs), are key signaling molecules that regulate a myriad of cellular processes. Specific interactions with PIPs are crucial for the functions of many signaling proteins, highlighting the need for a convenient and robust method to study lipid-protein interactions. Previously, we established a fluorescence microscopy-based lipid single-molecule pulldown (lipid-SiMPull) assay for detecting interactions between fluorescently tagged proteins of interest in whole-cell lysates and small unilamellar vesicles containing phospholipids of interest. Despite unique advantages of the lipid-SiMPull assay, small unilamellar vesicle is not an optimal membrane model due to its instability, heterogeneity in size, and a membrane curvature inconsistent with the relative flatness of the cell membrane. Here, we report the use of lipid Nanodiscs in lipid-SiMPull. Using PIP-protein pairs of known interactions, we show that Nanodiscs containing various PIPs can pull down protein targets specifically, with an estimated detection threshold of K_d in the 10-20 μM range. Remarkably, we find that each Nanodisc is bound by one copy of the protein (or protein dimer), conferring true single-molecule resolution to the assay. Transient interactions are characterized by the rebinding of proteins to individual Nanodiscs, and dissociation rates (k_off) are determined from dwell time analysis. We apply this assay to interrogate structural requirements for the stability of AKT binding of PI(3,4,5)P₃ and find that an intramolecular interaction between the PH domain and kinase domain is critical for stabilizing the AKT-PI(3,4,5)P₃ interaction. This work estalishes the Nanodisc SiMPull assay as a simple and powerful approach for investigating protein-lipid interactions with single-molecule resolution.</p>","PeriodicalId":16209,"journal":{"name":"Journal of Lipid Research","volume":" ","pages":"100846"},"PeriodicalIF":5.0,"publicationDate":"2025-06-20","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144368982","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Non-invasive identification of steatohepatitis in patients with MASLD using a sterol and lipidomic signature.","authors":"Ratna Budhi Pebriana, Ting Chen, Rico J E Derks, Niek Blomberg, Aldo Grefhorst, Yassene Mohammed, Max Nieuwdorp, Joanne Verheij, Michail Doukas, Patrick C N Rensen, Adriaan G Holleboom, Maarten E Tushuizen, Martin Giera","doi":"10.1016/j.jlr.2025.100845","DOIUrl":"10.1016/j.jlr.2025.100845","url":null,"abstract":"<p><p>The accumulation of cholesterol and other lipids leading to hepatic lipotoxicity drives the progression of metabolic dysfunction-associated steatotic liver (MASL) to metabolic dysfunction-associated steatohepatitis (MASH), the advanced progressive stage of metabolic dysfunction-associated steatotic liver disease (MASLD). For MASH diagnosis, liver biopsy remains the reference standard, despite its invasiveness and limitations. Thus, this study aimed to find blood-derived lipid markers for MASH. We investigated serum samples from 86 patients with histologically characterized MASLD, spanning the disease spectrum (i.e. 62 patients with MASL (Fibrosis grade 0-4) and 24 patients with MASH (Fibrosis grade 2-4) with a balanced distribution of hepatocellular carcinoma) and analyzed sterol composition and lipidome. To identify the presence of MASH, logistic regression was performed on each candidate either in a single or combination with various clinical parameters. Serum levels of desmosterol and phosphatidylcholine are increased in patients with MASH compared to those with MASL. After exclusion of patients using lipid lowering drugs, an increase was also found in serum levels of cholesterol, cholesterol ester, lysophosphatidylcholine, lysophosphatidylethanolamine, phosphatidylethanolamine, and several individual lipid species. The ROC curve of each lipid candidate show the potential use of desmosterol, phosphatidylcholine, and a panel of lipid species in combination with alanine aminotransferase as potential diagnostic markers, characterized by a respective AUROC of 0.79 (95% CI 0.66-0.92), 0.80 (95% CI 0.64-0.97), and 0.91 (95% CI 0.82-1.00). Serum sterol and lipidome markers are characterized by strong AUROC results to distinguish with high accuracy MASH from MASL, potentially paving the way for future MASH biomarker development.</p>","PeriodicalId":16209,"journal":{"name":"Journal of Lipid Research","volume":" ","pages":"100845"},"PeriodicalIF":5.0,"publicationDate":"2025-06-20","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144368983","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Optogenetic perturbation of lipid droplet localization affects lipid metabolism and development in Drosophila.","authors":"Xin Deng, Wei Wang, Dandan Peng, Luhao Zhang, Zhihao Ma, Junfen Fu, Chao Tong, Yingke Xu","doi":"10.1016/j.jlr.2025.100848","DOIUrl":"10.1016/j.jlr.2025.100848","url":null,"abstract":"<p><p>Lipid droplets (LDs) are dynamic organelles crucial for lipid storage and homeostasis. Despite extensive documentation of their importance, the causal relationship between LD localization and function in health and disease remains inadequately understood. Here, we developed optogenetics-based tools, termed \"Opto-LDs,\" which facilitate the interaction between LDs and motor proteins in a light-dependent manner, enabling precise control of LD localization within cells. Utilizing these optogenetic modules, we demonstrated that light-induced relocation of LDs to the periphery of hepatocytes results in elevated very-low-density lipoprotein (VLDL) secretion, recapturing the beneficial effect of insulin in vitro. Furthermore, our studies in transgenic Drosophila revealed that proper LD localization is critical for embryonic development, with mistargeting of LDs significantly affecting egg hatching success. In summary, our work underscores the great importance of LD localization in lipid metabolism and development, and our developed tools offer valuable insights into the functions of LDs in health and disease.</p>","PeriodicalId":16209,"journal":{"name":"Journal of Lipid Research","volume":" ","pages":"100848"},"PeriodicalIF":5.0,"publicationDate":"2025-06-20","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144368984","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Cyclodextrins inhibit TRPV1 and TRPA1 activation-induced nociception via cholesterol depletion.","authors":"Andrea Nehr-Majoros, Lajos Karakai, Maja Payrits, Noémi Bencze, Ágnes Kemény, György Sétáló, Rita Börzsei, Csaba Hetényi, Zsuzsanna Helyes, Éva Szőke","doi":"10.1016/j.jlr.2025.100844","DOIUrl":"10.1016/j.jlr.2025.100844","url":null,"abstract":"<p><p>The nociceptive Transient Receptor Potential Vanilloid 1 (TRPV1) and Ankyrin 1 (TRPA1) channels are predominantly expressed on peptidergic sensory nerves, being involved in pain sensation and neurogenic inflammation induced by local release of pro-inflammatory neuropeptides in the innervation area. Their activation is facilitated by cholesterol-rich lipid microdomains (lipid rafts) in the plasma membrane. Cyclodextrin (CD) derivatives deplete cholesterol from membrane rafts, reducing receptor activation in vitro, anticipating in vivo analgesic effects. We compared three different CD derivatives selected based on our previous results: random methylated β-cyclodextrin, (2-hydroxypropyl)-β-cyclodextrin and sulfobutylether-β-cyclodextrin. The effects of the topical CD pretreatments were evaluated in acute pain and neurogenic vasodilatation models in mice 30 min after TRPV1 (resiniferatoxin) or TRPA1 (formalin or mustard oil) receptor agonist administration. Intraplantar CD pretreatments significantly reduced the duration of nocifensive behaviors during the neurogenic inflammatory phase of the formalin test, as well as mechanical, but not thermal hyperalgesia following resiniferatoxin injection. CD-pretreatment significantly reduced mustard oil-induced acute neurogenic vasodilatation in the mouse ear and decreased the total cholesterol content in the plantar skin and ear tissues. Cholesterol depletion was restored by cholesterol loaded CDs. However, overloading cells with cholesterol did not significantly affect cholesterol depletion. In silico modeling showed that the methylated derivative RAMEB has different cholesterol binding mode compared to HPBCD and SBECD. We present the first in vivo results showing that these CD derivatives are promising agents for exerting peripheral analgesia and anti-inflammation via cholesterol depletion, also supported by our in vitro and in silico findings.</p>","PeriodicalId":16209,"journal":{"name":"Journal of Lipid Research","volume":" ","pages":"100844"},"PeriodicalIF":5.0,"publicationDate":"2025-06-16","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12274755/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144325987","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"The origin of hydroxy-cyclohexenone fatty acids from skin barrier protein and relevance to covalent binding of ceramides.","authors":"Saori Noguchi, William E Boeglin, Fumie Nakashima, Donald F Stec, M Wade Calcutt, Takuya Takeichi, Masashi Akiyama, Alan R Brash","doi":"10.1016/j.jlr.2025.100843","DOIUrl":"10.1016/j.jlr.2025.100843","url":null,"abstract":"<p><p>Lipid constituents of the skin permeability barrier include a portion of ceramides and fatty acids covalently bound to the barrier protein. The covalent binding requires enzymatic oxidation of linoleate (C18:2) esterified to skin-specific acylceramides, forming a reactive 9,10-epoxy-11E-13-keto derivative. Barrier proteins treated with alkali release the bound lipids and as described recently, including two prominent cyclic linoleate derivatives, C18 hydroxy-cyclohexenone fatty acids. Herein we addressed the origin of these cyclic products by alkali treatment of potential precursors. A UV-based assay indicated the rates of Michael adduction of 9,10-epoxy-11E-13-keto to cysteine are two orders of magnitude faster than for a typical unsaturated keto fatty acid, and 10-fold faster for the dihydroxy analog, rationalizing their biosynthesis for protein adduction. Alkali treatment degraded the epoxy-ketone and its cysteinyl (glutathione) adduct to multiple UV-absorbing products, although not including the hydroxy-cyclohexenones. By contrast, these derivatives were prominently produced from KOH treatment of the 9,10-dihydroxy-13-ketone or its glutathione adduct. As further evidence of the origin of the hydroxy-cyclohexenones, LC-MS quantitation showed a 90% reduction following KOH treatment of epidermis from mice deficient in Srd9c7, the dehydrogenase in the linoleate oxidation pathway. Taken together, the results confirm the hydroxy-cyclohexenones as derivatives of the linoleate oxidations in the skin barrier pathway and identify the dihydroxy-ketone as a component of the covalently-bonded lipids, and critical to integrity of the epidermal barrier.</p>","PeriodicalId":16209,"journal":{"name":"Journal of Lipid Research","volume":" ","pages":"100843"},"PeriodicalIF":5.0,"publicationDate":"2025-06-14","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12274931/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144310024","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Circulating membrane aminophospholipids contribute to thrombotic risk in rheumatoid arthritis.","authors":"Daniela O Costa, Majd B Protty, Victoria J Tyrrell, Ali A Hajeyah, Bethan H Morgan, Ben Mead, Martin Giera, Peter W Collins, P Vince Jenkins, Ernest Choy, Simon A Jones, Valerie B O'Donnell","doi":"10.1016/j.jlr.2025.100842","DOIUrl":"10.1016/j.jlr.2025.100842","url":null,"abstract":"<p><p>Patients with rheumatoid arthritis (RA) are at elevated risk of thrombotic events, yet the underlying mechanisms remain unknown. The contribution of the procoagulant membrane surface provided by aminophospholipids (aPLs) in driving thrombotic risk in RA has not been investigated. Specifically, neither the type of aPL exposed on circulating blood cell membranes in patients is characterized nor is their ability to support thrombin generation is known. Here, lipidomics was used to characterize the external-facing and total levels of aPL molecular species in RA, specifically phosphatidylserine and phosphatidylethanolamine on extracellular vesicles (EVs), platelets, and white blood cells (WBCs). The ability of the cells and EVs to support thrombin generation from patients and healthy controls was compared using an in vitro prothrombinase assay. RA patient plasma had significantly higher levels of thrombin-antithrombin and d-dimers, indicating increased thrombotic activity in vivo. Higher EV and platelet counts were seen in RA, but WBC counts were not elevated. EVs from RA patients supported higher levels of thrombin generation compared with healthy controls, whereas for platelets and WBC, thrombin generation was similar for both groups. EVs from RA patients also showed elevated external-facing phosphatidylserine molecular species, with total aPL also increased. For platelets and WBC, total and external-facing aPL levels were similar. Thrombin-antithrombin (TAT) complexes significantly correlated with EV particle counts, indicating that their circulating numbers are directly related to coagulation in vivo. Overall, our data suggest that elevated plasma EV levels in RA are a major source of procoagulant membranes, contributing to thrombotic risk in RA.</p>","PeriodicalId":16209,"journal":{"name":"Journal of Lipid Research","volume":" ","pages":"100842"},"PeriodicalIF":5.0,"publicationDate":"2025-06-14","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12273563/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144310022","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"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":"10.1016/j.jlr.2025.100841","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":5.0,"publicationDate":"2025-06-14","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12274753/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144310023","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}