Newton H. Tran , Dinesh Babu , Steven Lockhart , Andrew G. Morgan , Nadine Commandeur , Md Harunur Rashid , Béla Reiz , Lusine Tonoyan , Arno G. Siraki
{"title":"The oxidation of fenamic acid NSAIDs by neutrophil myeloperoxidase produces toxic reactive metabolites that induce leukemic cell death","authors":"Newton H. Tran , Dinesh Babu , Steven Lockhart , Andrew G. Morgan , Nadine Commandeur , Md Harunur Rashid , Béla Reiz , Lusine Tonoyan , Arno G. Siraki","doi":"10.1016/j.rbc.2023.100013","DOIUrl":"https://doi.org/10.1016/j.rbc.2023.100013","url":null,"abstract":"<div><p>Fenamic acids are a group of non-steroidal anti-inflammatory drugs (NSAIDs) that are among the most common drugs prescribed globally. However, they have been associated with many adverse effects, such as agranulocytosis, neutropenia, hepatotoxicity, and nephrotoxicity. The interactions between peroxidase enzymes and fenamic acid-like NSAIDs cause the formation of reactive species, potentially involved in side effects. The aim of this study was to investigate the neutrophil myeloperoxidase (MPO)-mediated bioactivation of fenamic acids based on N-phenylanthranilic acid (NPA) and its four drug analogues: flufenamic acid (FFA), mefenamic acid (MFA), meclofenamic acid (MCFA), and tolfenamic acid (TFA). We hypothesized that the enzymatic oxidation of fenamic acids by MPO/hydrogen peroxide (H<sub>2</sub>O<sub>2</sub>) would produce reactive metabolites, cause oxidative damage and induce cytotoxicity. We utilized UV–Vis spectrophotometry, liquid chromatography-mass spectrometry (LC-MS), and electron paramagnetic spin resonance (EPR) spectroscopy using purified MPO from human neutrophils. In addition, <em>in vitro</em> studies were performed with MPO-containing human promyelocytic leukemia (HL-60) cells for cytotoxicity and immuno-spin trapping to detect protein-free radicals. UV–Vis spectrophotometry revealed that MPO oxidized the fenamic acids. LC-MS analyses revealed the formation of dimers, hydroxylated, and quinoneimine species, and glutathione (GSH) conjugates. EPR spin trapping with DMPO using GSH revealed that fenamic acids produced glutathionyl radicals in a concentration-dependent manner. We also detected the formation of protein-free radicals in HL-60 cells, which correlated with cytotoxicity. Despite the minor structural differences between the fenamic acids, there were variations in their oxidation potential. These findings revealed a correlation between pro-oxidant metabolite reactivity and cytotoxicity caused by fenamic acid NSAIDs.</p></div>","PeriodicalId":101065,"journal":{"name":"Redox Biochemistry and Chemistry","volume":"5 ","pages":"Article 100013"},"PeriodicalIF":0.0,"publicationDate":"2023-08-12","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"49754463","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
{"title":"Inter(pseudo)halogens with relevance to peroxidase-mediated reactions","authors":"Jürgen Arnhold , Ernst Malle","doi":"10.1016/j.rbc.2023.100012","DOIUrl":"https://doi.org/10.1016/j.rbc.2023.100012","url":null,"abstract":"<div><p>In mammals, heme peroxidases are well known to generate oxidized (pseudo)halide products such as hypochlorous acid, hypobromous acid, oxidized iodine species, and hypothiocyanite. In addition, inter(pseudo)halogens are also oxidized (pseudo)halide compounds where two or more different (pseudo)halides are combined within a molecule without participation of other atoms. However, the information of this group of chemicals as potential products of peroxidases is limited and very fragmentary. In this review, we summarize current knowledge about chemical properties of inter(pseudo)halogens, their role as products of peroxidase-mediated conversions, and possible applications of these compounds in antimicrobial defense. The major focus is directed on bromyl chloride, cyanogen halides, and some products derived from interaction of oxidized iodine with thiocyanate.</p></div>","PeriodicalId":101065,"journal":{"name":"Redox Biochemistry and Chemistry","volume":"5 ","pages":"Article 100012"},"PeriodicalIF":0.0,"publicationDate":"2023-08-07","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"49726364","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Reagan M. McGuffee , Christy M. Hadfield , David A. Ford
{"title":"Lipid biology of plasmalogen-derived halolipids: Signature molecules of myeloperoxidase and eosinophil peroxidase activity","authors":"Reagan M. McGuffee , Christy M. Hadfield , David A. Ford","doi":"10.1016/j.rbc.2023.100011","DOIUrl":"https://doi.org/10.1016/j.rbc.2023.100011","url":null,"abstract":"<div><p>Myeloperoxidase and eosinophil peroxidase exert their antimicrobial functions through the oxidative actions of their hypohalous acid products. Plasmalogen phospholipids are particularly susceptible to oxidation of their vinyl ether functional group by hypohalous acids. This produces a family of halogenated lipid products with pro-inflammatory roles and potential biomarker utility. The initial product of plasmalogen oxidation by HOCl is 2-chlorofatty aldehyde, which has been shown to play a key role at the blood-endothelium interface. <em>In vitro</em> and <em>in vivo</em> studies indicate increased endothelial barrier permeability, neutrophil chemotaxis, neutrophil and platelet adherence to endothelium, and promotion of erythrocyte lysis as some of its effects. These effects may be due to protein modification by 2-chlorofatty aldehyde. 2-Chlorofatty aldehyde is metabolized by host dehydrogenases to 2-chlorofatty acid. While it is less chemically reactive, 2-chlorofatty acid has partial overlap of pro-inflammatory effects with 2-chlorofatty aldehyde and unique actions such as induction of neutrophil extracellular trap formation. The stability of 2-chlorofatty acid in plasma also makes it well-suited as a biomarker of HOCl generation, and its plasma levels may be predictive of disease outcomes. 2-Bromofatty aldehydes and acids are produced analogously from HOBr reaction with plasmalogens. Their functions have yet to be well-elucidated, though similarities with chlorolipids have been observed, and increased reactivity with proteins is expected through enhanced electrophilicity of the alpha carbon. Altogether, these halogenated lipids represent underexplored mediators of diseases involving excess hypohalous acid production.</p></div>","PeriodicalId":101065,"journal":{"name":"Redox Biochemistry and Chemistry","volume":"5 ","pages":"Article 100011"},"PeriodicalIF":0.0,"publicationDate":"2023-08-02","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"49738430","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
{"title":"Electron paramagnetic resonance (EPR) for investigating relevant players of redox reactions: Radicals, metalloproteins and transition metal ions","authors":"Ohara Augusto, Daniela Ramos Truzzi, Edlaine Linares","doi":"10.1016/j.rbc.2023.100009","DOIUrl":"https://doi.org/10.1016/j.rbc.2023.100009","url":null,"abstract":"<div><p>Electron paramagnetic resonance (EPR) spectroscopy is unique in providing robust information about free radicals, transition metal ions and metalloenzymes, which are crucial players in redox processes. EPR had a major role in advancing the redox biology field during the 20th century, but the interest in this methodology considerably decreased in recent years. Here, we discuss potential reasons for this decline as well as potential reasons for maintaining the mind open to the many possibilities brought by EPR and associated methodologies to the redox field. We present the fundamentals of EPR using pictorial images and minimal physicochemical language. We also present EPR derived methodologies developed to detect radical metabolites, that is, direct EPR of solutions (static and continuous-flow), direct EPR of frozen solutions, spin-trapping and spin-scavenging, showing examples and discussing the advantages and drawbacks of each one. Finally, we discuss the EPR spectra of metalloproteins and metal ion complexes of biological interest, which are more complex than those of radical metabolites in solution. In addition to introduce EPR methodologies to those new to the redox field, our goal is to show that these methodologies can contribute to advance the field.</p></div>","PeriodicalId":101065,"journal":{"name":"Redox Biochemistry and Chemistry","volume":"5 ","pages":"Article 100009"},"PeriodicalIF":0.0,"publicationDate":"2023-07-29","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"49738239","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Els A. Hartsema, Line A.E. Hallberg, Kristine Barlous, Clare L. Hawkins
{"title":"Modification of histones by the myeloperoxidase-derived oxidant hypochlorous acid (HOCl) alters their reactivity with vascular smooth muscle cells","authors":"Els A. Hartsema, Line A.E. Hallberg, Kristine Barlous, Clare L. Hawkins","doi":"10.1016/j.rbc.2023.100010","DOIUrl":"https://doi.org/10.1016/j.rbc.2023.100010","url":null,"abstract":"<div><p>In the nucleus, histones are essential in the packaging of DNA and the regulation of gene expression. These histones can also be released to the extracellular space by mechanisms such as necrosis and neutrophil extracellular trap (NET) formation. Histones are cytotoxic and cause sterile inflammation, and as a result, have been implicated in tissue damage in several pathologies, including atherosclerosis. Myeloperoxidase (MPO) is also present on NETs, which is catalytically active and able to produce hypochlorous acid (HOCl). This could modify histones and alter their extracellular reactivity. In this study, we compared the reactivity of histones with and without modification by HOCl with primary human coronary artery smooth muscle cells (HCASMCs). Histones induced a loss in viability and cell death primarily by apoptosis, which was attenuated on modification of the histones by HOCl. Exposure of HCASMCs to histones also resulted in the increased expression of the pro-inflammatory genes monocyte chemoattractant protein-1 (MCP-1), interleukin 6 (IL-6), and vascular cell adhesion molecule-1 (VCAM-1) and a decrease in intracellular thiols. In addition, there were changes in the expression of the stress related gene heme oxygenase-1 (HO-1). Modification of the histones with HOCl had no significant influence on changes in gene expression or thiol loss, in contrast to the cytotoxicity studies. Together, these studies provide new insight into the pathways by which histones could promote vascular dysfunction, which could be relevant to inflammatory diseases, such as atherosclerosis and sepsis, which are associated with elevated NET release and high circulating histones, respectively.</p></div>","PeriodicalId":101065,"journal":{"name":"Redox Biochemistry and Chemistry","volume":"5 ","pages":"Article 100010"},"PeriodicalIF":0.0,"publicationDate":"2023-07-29","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"49738241","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Katja E. Menger , Angela Logan , Ulrich F.O. Luhmann , Alexander J. Smith , Alan F. Wright , Robin R. Ali , Michael P. Murphy
{"title":"In vivo measurement of mitochondrial ROS production in mouse models of photoreceptor degeneration","authors":"Katja E. Menger , Angela Logan , Ulrich F.O. Luhmann , Alexander J. Smith , Alan F. Wright , Robin R. Ali , Michael P. Murphy","doi":"10.1016/j.rbc.2023.100007","DOIUrl":"https://doi.org/10.1016/j.rbc.2023.100007","url":null,"abstract":"<div><p>Retinitis pigmentosa (RP) is a disease characterised by photoreceptor cell death. It can be initiated by mutations in a number of different genes, primarily affecting rods, which will die first, resulting in loss of night vision. The secondary death of cones then leads to loss of visual acuity and blindness. We set out to investigate whether increased mitochondrial reactive oxygen species (ROS) formation, plays a role in this sequential photoreceptor degeneration. To do this we measured mitochondrial H<sub>2</sub>O<sub>2</sub> production within mouse eyes <em>in vivo</em> using the mass spectrometric probe MitoB. We found higher levels of mitochondrial ROS that preceded photoreceptor loss in four mouse models of RP: <em>Pde6b</em><sup><em>rd1/rd1</em></sup><em>; Prhp2</em><sup><em>rds/rds</em></sup><em>; RPGR</em><sup><em>−/−</em></sup><em>; Cln6</em><sup><em>nclf</em></sup>. In contrast, there was no increase in mitochondrial ROS in loss of function models of vision loss (<em>GNAT</em><sup><em>−/−</em></sup><em>, OGC</em>), or where vision loss was not due to photoreceptor death (<em>Cln3</em>). Upregulation of <em>Nrf2</em> transcriptional activity with dimethylfumarate (DMF) lowered mitochondrial ROS in <em>RPGR</em><sup><em>−/−</em></sup> mice. These findings have important implications for the mechanism and treatment of RP.</p></div>","PeriodicalId":101065,"journal":{"name":"Redox Biochemistry and Chemistry","volume":"5 ","pages":"Article 100007"},"PeriodicalIF":0.0,"publicationDate":"2023-07-27","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"49726776","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Martina Paumann-Page , Louisa V. Ashby , Irada Khalilova , Nicholas J. Magon , Stefan Hofbauer , Louise N. Paton , Paul G. Furtmüller , Christian Obinger , Anthony J. Kettle
{"title":"Hypochlorous acid inactivates myeloperoxidase inside phagocytosing neutrophils","authors":"Martina Paumann-Page , Louisa V. Ashby , Irada Khalilova , Nicholas J. Magon , Stefan Hofbauer , Louise N. Paton , Paul G. Furtmüller , Christian Obinger , Anthony J. Kettle","doi":"10.1016/j.rbc.2023.100008","DOIUrl":"https://doi.org/10.1016/j.rbc.2023.100008","url":null,"abstract":"<div><p>When neutrophils phagocytose bacteria, they release myeloperoxidase (MPO) into phagosomes to catalyse the conversion of superoxide to the potent antimicrobial oxidant hypochlorous acid (HOCl). Here we show that within neutrophils, MPO is inactivated by HOCl. In this study, we aimed to identify the effects of HOCl on the structure and function of MPO, and determine the enzyme's susceptibility to oxidative inactivation during phagocytosis. When hydrogen peroxide was added to a neutrophil granule extract containing chloride, MPO activity was rapidly lost in a HOCl-dependent reaction. With high concentrations of hydrogen peroxide, western blotting demonstrated that MPO was both fragmented and converted to high molecular weight aggregates. Using the purified enzyme, we showed that HOCl generated by MPO inactivated the enzyme by destroying its prosthetic heme groups and releasing iron. MPO protein was additionally modified by forming high molecular weight aggregates. Before inactivation occurred, MPO chlorinated itself to convert most of its amine groups to dichloramines. When human neutrophils phagocytosed <em>Staphylococcus aureus</em>, they released MPO that was largely inactivated in a process that required production of superoxide. Enzyme inactivation occurred inside neutrophils because it was not blocked when extracellular HOCl was scavenged with methionine. The inactivated enzyme contained a chlorinated tyrosine residue, establishing that it had reacted with HOCl. Our results demonstrate that MPO will substantially inactivate itself during phagocytosis, which may limit oxidant production inside phagosomes. Other neutrophil proteins are also likely to be inactivated. The chloramines formed on neutrophil proteins may contribute to the bactericidal milieu of the phagosome.</p></div>","PeriodicalId":101065,"journal":{"name":"Redox Biochemistry and Chemistry","volume":"5 ","pages":"Article 100008"},"PeriodicalIF":0.0,"publicationDate":"2023-07-26","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"49726740","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
{"title":"Could CO2 be a player in a redox relay team?","authors":"Alexander V. Peskin","doi":"10.1016/j.rbc.2023.100006","DOIUrl":"https://doi.org/10.1016/j.rbc.2023.100006","url":null,"abstract":"<div><p>It has been established that hydrogen peroxide (H<sub>2</sub>O<sub>2</sub>) acts as a signalling messenger by triggering the reversible oxidation of redox-regulated proteins via a redox-relay provided by peroxiredoxins (Prdxs). Exceptionally high reactivity of Prdxs with H<sub>2</sub>O<sub>2</sub> exceeding other thiols by orders of magnitude places Prdxs as sensors of H<sub>2</sub>O<sub>2</sub> and distributers of oxidizing equivalents to specific thiol targets which can't be oxidized by H<sub>2</sub>O<sub>2</sub> directly. By this mechanism the oxidative stress response can be achieved.</p><p>Despite its involvement in oxidative stress responses, H<sub>2</sub>O<sub>2</sub> is continuously generated as a normal metabolite necessary for regular cell functioning. The challenge lies in understanding how the Prdx-dependent redox relay can differentiate between basal levels of H<sub>2</sub>O<sub>2</sub> and excessive amounts that lead to oxidative stress.</p><p>Peroxymonocarbonate, an oxidant formed when H<sub>2</sub>O<sub>2</sub> reacts with CO<sub>2</sub>/HCO<sub>3</sub><sup>−</sup>, emerges as a potent cellular oxidant. The peroxymonocarbonate formation could be catalysed and then consumed at localised sites by certain thiol proteins. This mechanism could prevent H<sub>2</sub>O<sub>2</sub> from reacting with Prdx, thereby averting the redox-relayed activation of regulatory thiol proteins and subsequent oxidative stress response below a certain level of H<sub>2</sub>O<sub>2</sub>.</p></div>","PeriodicalId":101065,"journal":{"name":"Redox Biochemistry and Chemistry","volume":"5 ","pages":"Article 100006"},"PeriodicalIF":0.0,"publicationDate":"2023-07-14","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"49726774","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Kailash Manda , Kei Ohkubo , Yoshimi Shoji , A. K. M. Raushan Kabir Zoardar , Masato Kamibayashi , Toshihiko Ozawa , Kazunori Anzai , Ikuo Nakanishi
{"title":"In vitro radical-scavenging mechanism of melatonin and its in vivo protective effect against radiation-induced lipid peroxidation","authors":"Kailash Manda , Kei Ohkubo , Yoshimi Shoji , A. K. M. Raushan Kabir Zoardar , Masato Kamibayashi , Toshihiko Ozawa , Kazunori Anzai , Ikuo Nakanishi","doi":"10.1016/j.rbc.2023.100003","DOIUrl":"https://doi.org/10.1016/j.rbc.2023.100003","url":null,"abstract":"<div><p>Melatonin (<em>N</em>-acetyl-5-methoxytryptamine, MLT), an evolutionarily conserved indoleamine, is known to act as an antioxidant. However, the evidence indicating the role of MLT as a powerful chain-breaking antioxidant by scavenging peroxyl radical remains controversial. The radical-scavenging rate of MLT determined in this study in methanol using galvinoxyl radical (GO<sup>•</sup>) was much lower than that of an α-tocopherol model compound. The acceleration of the GO<sup>•</sup>-scavenging reaction by MLT was observed in the presence of magnesium ion (Mg<sup>2+</sup>), a bio-related redox-inactive metal ion, suggesting that this reaction may proceed via a rate-determining electron transfer followed by proton transfer. The coordination of Mg<sup>2+</sup> to the carbonyl oxygen in the one-electron reduced species of GO<sup>•</sup> (GO<sup>–</sup>) may stabilize the product, resulting in the acceleration of the electron-transfer process. We also demonstrated that prophylactically administrated MLT efficiently inhibited the lipid peroxide-derived protein modification, which can be detected by a sensitive marker, <em>N</em><sup>ε</sup>-(hexanoyl)lysine adduct, in the plasma of X-irradiated mice. The relatively weak GO<sup>•</sup>-scavenging activity of MLT suggests that the ameliorative effect of MLT against in vivo lipid peroxidation does not result from the direct scavenging of lipid peroxyl radicals by MLT. Therefore, the observed superior protective efficiency of MLT against in vivo lipid peroxidation may partly support the earlier studies, which reported the synergistic antioxidative effect of the metabolites of MLT.</p></div>","PeriodicalId":101065,"journal":{"name":"Redox Biochemistry and Chemistry","volume":"3 ","pages":"Article 100003"},"PeriodicalIF":0.0,"publicationDate":"2023-07-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"49750806","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Rosangela S. Santos , Márcia S.F. Franco , Felipe G. Ravagnani , Adriano B. Chaves-Filho , Sayuri Miyamoto , Mauricio S. Baptista , Mikhail S. Shchepinov , Marcos Y. Yoshinaga
{"title":"Intracellular distribution of bis-allylic deuterated linoleic acid into the lipidome of human keratinocytes","authors":"Rosangela S. Santos , Márcia S.F. Franco , Felipe G. Ravagnani , Adriano B. Chaves-Filho , Sayuri Miyamoto , Mauricio S. Baptista , Mikhail S. Shchepinov , Marcos Y. Yoshinaga","doi":"10.1016/j.rbc.2023.100005","DOIUrl":"https://doi.org/10.1016/j.rbc.2023.100005","url":null,"abstract":"<div><p>Polyunsaturated fatty acids (PUFA) are particularly susceptible to free radical-induced lipid peroxidation (LPO). Specific deuteration at bis-allylic positions of PUFA (D-PUFA) has been recently proposed as a way to inhibit the LPO. Here, a high mass resolution untargeted lipidomic analysis protocol was applied to examine the changes in the lipidome of keratinocytes supplemented with bis-allylic deuterated linoleic acid (D<sub>2</sub>-LA). Incorporation of D<sub>2</sub>-LA occurs preferentially in membrane phospholipids such as phosphatidylcholine and phosphatidylethanolamine, followed by triglycerides. However, the relative contribution of D<sub>2</sub>-LA among membrane lipids is highest in cardiolipin (60%) followed by its precursor phosphatidylglycerol (50%). Cardiolipins are enriched in PUFA and exclusively located in mitochondrial membranes, thus representing major targets for lipid peroxidation. These findings indicate that D<sub>2</sub>-LA supplementation is linked to the preservation of mitochondrial function under oxidative stress. Finally, our study highlights the suitability of high mass resolution lipidomic analysis to investigate lipid metabolism at the level of individual molecular species in stable isotope tracing experiments.</p></div>","PeriodicalId":101065,"journal":{"name":"Redox Biochemistry and Chemistry","volume":"5 ","pages":"Article 100005"},"PeriodicalIF":0.0,"publicationDate":"2023-06-16","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"49726689","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}