Prideapplication/xmlftp://ftp.pride.ebi.ac.uk/pride/data/archive/2026/05/PXD075585/20260306_171816_20260217-0022_Report.tsvftp://ftp.pride.ebi.ac.uk/pride/data/archive/2026/05/PXD075585/SampleLabels.txtftp://ftp.pride.ebi.ac.uk/pride/data/archive/2026/05/PXD075585/checksum.txtftp://ftp.pride.ebi.ac.uk/pride/data/archive/2026/05/PXD075585/20260218-0028_AS2_003381_250216_IVAN_MRL_HepKati_24.rawftp://ftp.pride.ebi.ac.uk/pride/data/archive/2026/05/PXD075585/20260217-1747_AS2_003376_250216_IVAN_MRL_HepKati_19.rawftp://ftp.pride.ebi.ac.uk/pride/data/archive/2026/05/PXD075585/20260217-1346_AS2_003373_250216_IVAN_MRL_HepKati_16.rawftp://ftp.pride.ebi.ac.uk/pride/data/archive/2026/05/PXD075585/20260217-0543_AS2_003367_250216_IVAN_MRL_HepKati_10.rawftp://ftp.pride.ebi.ac.uk/pride/data/archive/2026/05/PXD075585/20260217-1626_AS2_003375_250216_IVAN_MRL_HepKati_18.rawftp://ftp.pride.ebi.ac.uk/pride/data/archive/2026/05/PXD075585/20260217-2148_AS2_003379_250216_IVAN_MRL_HepKati_22.rawftp://ftp.pride.ebi.ac.uk/pride/data/archive/2026/05/PXD075585/20260217-1907_AS2_003377_250216_IVAN_MRL_HepKati_20.rawftp://ftp.pride.ebi.ac.uk/pride/data/archive/2026/05/PXD075585/20260217-1506_AS2_003374_250216_IVAN_MRL_HepKati_17.rawftp://ftp.pride.ebi.ac.uk/pride/data/archive/2026/05/PXD075585/20260217-0303_AS2_003365_250216_IVAN_MRL_HepKati_8.rawftp://ftp.pride.ebi.ac.uk/pride/data/archive/2026/05/PXD075585/20260217-2027_AS2_003378_250216_IVAN_MRL_HepKati_21.rawftp://ftp.pride.ebi.ac.uk/pride/data/archive/2026/05/PXD075585/20260217-0423_AS2_003366_250216_IVAN_MRL_HepKati_9.rawftp://ftp.pride.ebi.ac.uk/pride/data/archive/2026/05/PXD075585/20260217-0022_AS2_003363_250216_IVAN_MRL_HepKati_6.rawftp://ftp.pride.ebi.ac.uk/pride/data/archive/2026/05/PXD075585/20260217-2308_AS2_003380_250216_IVAN_MRL_HepKati_23.rawftp://ftp.pride.ebi.ac.uk/pride/data/archive/2026/05/PXD075585/20260217-0142_AS2_003364_250216_IVAN_MRL_HepKati_7.rawprimaryOK200grevengoed@sund.ku.dkIvan BradićMass SpectrometryBottom-up proteomicsFatty acid oxidationEndogenous lipid metaboliteArachidonic acidN-acyl taurineMasldAra-thttps://www.ebi.ac.uk/pride/archive/projects/PXD075585HepatocyteSamples were subjected to lysis in 4% sodium dodecyl sulfate (SDS) and 100 mM Tris-HCl pH 8.0 with a 1x Complete protease inhibitor cocktail (EDTA-free, Roche, catalog number 118735800001) and a 1x PhosSTOP phosphatase inhibitor (Sigma, catalog number 0490683700. The samples were sonicated with 1 second on and 1 second off method for 2 minutes at an amplitude of 40%. Following this, the lysates underwent denaturation at 95 °C for five minutes. The total protein levels were determined through a bicinchoninic acid (BCA) assay. Aliquots of 100 µg of protein per sample were reduced with 10 mM dithiothreitol (DTT) and alkylated with 20 mM iodoacetamide (IAA). Sample digestion was performed using the Protein Aggregation Capture (PAC) method (Batth et al., 2019; PMID: 30833379), automated on a KingFisher Flex system (Thermo Fisher Scientific, Waltham, MA) using an adjusted protocol provided by ReSyn Biosciences (Gauteng, South Africa). For the automated workflow, a series of 96-well plates were prepared. A "capture" plate (Plate 1) was prepared by mixing 100 µg of protein lysate with 400 µg of MagReSyn® Hydroxyl beads (ReSyn Biosciences), with acetonitrile added to a final concentration of 70%. Three subsequent wash plates (Plates 2, 3, and 4) were prepared containing 1 mL of 100% acetonitrile per well, followed by two additional wash plates (Plates 5 and 6) containing 1 mL of 70% ethanol per well. Finally, an "on-bead digest" plate (Plate 8) was prepared with 250 µL of 50 mM ammonium bicarbonate (pH 8.0), supplemented with 0.04 AU/mg protein LysC and 5% (w/w) trypsin per well. After loading the prepared plates and a plastic comb into the KingFisher Flex robot, the system facilitated protein aggregation onto the beads in the capture plate. Following digestion, the beads were magnetically transferred to a new elution plate containing 1% trifluoroacetic acid (TFA). The newly eluted peptides were combined with the digested peptides remaining in Plate 8. The acidified peptide mixture was vacuum-dried and resuspended in 0.1% TFA prior to mass spectrometry analysis. Peptide separation was performed using a trap-and-elute configuration on a Vanquish™ Neo UHPLC system (Thermo Scientific™) coupled to an Orbitrap Astral mass spectrometer. Approximately 1 µg of peptides was loaded onto a PepMap™ Neo 5 µm C18 trap cartridge (300 µm × 5 mm; cat. no. 174500) and separated on an Easy-Spray™ PepMap™ Neo 2 µm C18 analytical column (75 µm × 500 mm; cat. no. ES75500PN). Chromatographic separation was achieved using a custom gradient of Solvent B (95% acetonitrile, 0.1% formic acid). The gradient was initiated at a constant 3% Solvent B with a flow rate of 500 nL/min for 2.5 minutes, after which the flow rate was reduced to 250 nL/min for 0.5 minutes while maintaining 3% Solvent B. This was followed by a steady increase to 25% Solvent B over 50 minutes, and a further increase to 35% Solvent B over the next 10 minutes. The column was then washed by ramping up to 95% Solvent B over 0.4 minutes. For the final wash and equilibration steps, the flow rate was increased to 350 nL/min over 0.1 minutes and held constant at 95% Solvent B for 6.5 minutes. The Orbitrap Astral mass spectrometer was operated in positive ion mode. MS1 survey scans were acquired in the Orbitrap at a resolution of 240,000, with a scan range of 380–980 m/z and a maximum injection time of 5 ms. Data-independent acquisition (DIA) MS2 scans were performed using higher-energy collision-induced dissociation (HCD) set to a normalized collision energy of 25%. MS2 fragment ions were detected using the Astral detector with a scan range of 150–2000 m/z, a maximum injection time of 5 ms, and a cycle time of 0.6 s.PrideRaw data files were searched with Spectronaut (20.3) using the Mus musculus reference proteome FASTA file, ID UP000000589, containing 21,757 genes (downloaded on 27.01.2025). Most search settings were kept at their defaults. In brief, search settings included trypsin-digested peptides with a maximum of 2 missed cleavages, carbamidomethyl (C) as a fixed modification, and a maximum of 5 variable modifications, including acetyl (Protein N-term) and oxidation (M). The Deep DirectDIA+ workflow was used for identification. For quantification, precursor filtering was set to “Identified (Qvalue)”, and no imputation was performed. The proteotypicity filter was set to “Only protein group specific”, and cross-run normalization was used.ProteomicsTrisha Jean GrevengoedDepartment of Biomedical Sciences, University of CopenhagenPARTIALMus Musculus (mouse)ibradic95@gmail.com10.1016/J.JHEP.2026.04.033University of Southern DenmarkDenmarkfalseArachidonoyl-taurine protects from diet-induced hepatic steatosis and inflammation -Metabolic dysfunction-associated steatotic liver disease and its progressive and inflamma-tory form metabolic dysfunction-associated steatohepatitis represents a global health challenge, with limited treatment options available. In this study we identified an endoge-nous, understudied omega-6 fatty acid metabolite, arachidonoyl-taurine (ARA-T), capable of mitigating liver disease. ARA-T levels increased in human plasma of chronic and acute fatty liver and their abundance can be driven in humans and mice by dietary supplementa-tion of arachidonic acid. Surprisingly, our genetic model of elevated circulating ARA-T levels prevented inflammation and hepatic steatosis by increased uptake and turnover of fatty ac-ids in the liver. Pharmacological administration of ARA-T reduced liver weight and diet-induced hepatic lipid deposition in mice, demonstrating its potential to protect against and reverse the progression of liver disease. Thus, ARA-T may represent a way to protect against pro-inflammatory actions of omega-6 fatty acids thereby contributing to regulation of in-flammation and accumulation of hepatic lipids.2026-05-202026-03-13PXD075585NEWT:6945NEWT:3555NEWT:241368NEWT:2NEWT:157546NEWT:190802NEWT:35554NEWT:150475NEWT:9417NEWT:347515NEWT:1216979NEWT:307972NEWT:32046NEWT:544496NEWT:5180NEWT:256737NEWT:115104NEWT:1081927NEWT:67825NEWT:13076NEWT:1249668NEWT:317NEWT:1736309NEWT:7227NEWT:7469NEWT:885318NEWT:4081NEWT:876138NEWT:554NEWT:98334NEWT:237561NEWT:10036NEWT:7574NEWT:1351NEWT:7215NEWT:272563NEWT:507601NCBITaxon:79857NCBITaxon:6157NEWT:95648NEWT:746360NEWT:6239NEWT:1589NEWT:470150NEWT:135622NEWT:216257NEWT:6915NEWT:9986NEWT:101510NEWT:4054NEWT:3880NEWT:8782NEWT:1000589NEWT:1902NEWT:85962NEWT:160488NEWT:28104NEWT:317447NEWT:7955NCBITaxon:2NEWT:985076NEWT:7959NEWT:2261NEWT:4565NEWT:1264690NEWT:6192NEWT:28532NCBITaxon:38727NEWT:34305NEWT:59729NCBITaxon:183674NEWT:224308NEWT:626528NEWT:139927NEWT:4558NEWT:209285NEWT:216595NEWT:243230NEWT:8355NEWT:931281NEWT:7029NEWT:1283300NEWT:334747NEWT:61235NCBITaxon:79824NCBITaxon:4563NEWT:5755NEWT:3218NEWT:5759NEWT:1736231NEWT:436486NEWT:6287NEWT:2242NEWT:300641NEWT:4784NEWT:727NEWT:9796NEWT:725NEWT:360106NEWT:260707NEWT:287NEWT:10117NEWT:10239NCBITaxon:6191NEWT:10116NEWT:1280NEWT:1836NEWT:1735272NEWT:83334NEWT:83332NEWT:29760NEWT:260704NEWT:703612NEWT:260705NEWT:80863NEWT:44685NEWT:2697049NEWT:1148NEWT:11676NEWT:55571NEWT:100226NCBITaxon:6073NEWT:4530NEWT:4896NEWT:6279NEWT:1123869NEWT:7370NEWT:83906NEWT:6282NEWT:1134506NEWT:575584NEWT:1773NEWT:38783NEWT:8727NEWT:1895NEWT:1182590NEWT:8726NEWT:10090NEWT:935293NEWT:749200NEWT:4120NEWT:5693NEWT:8724NEWT:51511NEWT:92867NEWT:8723NEWT:990346NEWT:5334NEWT:145953NEWT:257309NEWT:230741NEWT:284812NCBITaxon:10359NCBITaxon:1313NEWT:43330NEWT:242619NEWT:44544NEWT:632957NEWT:373995NEWT:5689NEWT:544404NEWT:3702NEWT:129249NEWT:8839NEWT:4232NEWT:990119NEWT:2758385NEWT:4113NEWT:837NEWT:11298NEWT:171101NEWT:196627NEWT:408172NEWT:5691NEWT:408170NEWT:493760NEWT:260710NEWT:627025NEWT:400772NEWT:1097677NEWT:3708NEWT:106592NEWT:1117957NEWT:9913NEWT:1432138NEWT:10312NEWT:4100NEWT:1076NEWT:6763NEWT:803NEWT:8030NEWT:29722NEWT:380394NEWT:1692259NEWT:1639NEWT:188229NEWT:3818NEWT:480NEWT:4909NEWT:180066NEWT:67767NEWT:46835NEWT:135588NEWT:1843183NEWT:95486NEWT:58002NEWT:9103NEWT:4577NEWT:5664NEWT:2157NEWT:146479NEWT:10306NEWT:1911079NEWT:8022NEWT:145943NEWT:3635NEWT:5811NEWT:235443NEWT:1480154NEWT:1274414NEWT:59202NEWT:3197NEWT:9615NEWT:10299NEWT:860688NEWT:884019NEWT:169963NEWT:36329NEWT:72407NEWT:9606NEWT:367830NEWT:157295NEWT:410289NEWT:373153NEWT:915099NEWT:74940NEWT:1450511NEWT:470NEWT:84023NEWT:9838NCBITaxon:9615NEWT:1193501NEWT:3055NEWT:6326NEWT:6689NEWT:2762NEWT:5476NEWT:1174673NEWT:562NEWT:33952NEWT:1274432NEWT:1274426NEWT:1423NEWT:4932NEWT:70448NEWT:9825NEWT:1274423NEWT:3603NEWT:698936NEWT:2759NEWT:3847NEWT:39946NEWT:9823NEWT:9940NEWT:573NEWT:9031NEWT:1274420NEWT:7091