MetaboLights

application/xml

ftp://ftp.ebi.ac.uk/pub/databases/metabolights/studies/public/MTBLS14085/m_MTBLS14085_LC-MS_negative_reverse-phase_metabolite_profiling_v2_maf.tsvftp://ftp.ebi.ac.uk/pub/databases/metabolights/studies/public/MTBLS14085/m_MTBLS14085_LC-MS_positive_reverse-phase_metabolite_profiling_v2_maf.tsvftp://ftp.ebi.ac.uk/pub/databases/metabolights/studies/public/MTBLS14085/a_MTBLS14085_LC-MS_positive_reverse-phase_metabolite_profiling.txtftp://ftp.ebi.ac.uk/pub/databases/metabolights/studies/public/MTBLS14085/s_MTBLS14085.txtftp://ftp.ebi.ac.uk/pub/databases/metabolights/studies/public/MTBLS14085/a_MTBLS14085_LC-MS_negative_reverse-phase_metabolite_profiling.txtftp://ftp.ebi.ac.uk/pub/databases/metabolights/studies/public/MTBLS14085/i_Investigation.txtprimaryOK200ftp://ftp.ebi.ac.uk/pub/databases/metabolights/studies/public/MTBLS14085Metabolites were identified by matching MS/MS spectra against the LipidBlast, HMDB, or in-house database using metaX software. Identification criteria included: mass tolerance < 10 ppm for precursor ions, and MS/MS spectral similarity score > 70%. Lipid species were annotated according to LIPID MAPS classification.MetaboLightsPublicLiquid Chromatography MS - negative - reverse-phaseLiquid Chromatography MS - positive - reverse-phaseInstrument: ACQUITY UPLC System (Waters Corporation, Milford, MA, USA)Column: ACQUITY UPLC CSH C18 column (1.7 μm, 100 mm × 2.1 mm, Waters, UK)Column Temperature: 40°CFlow Rate: 0.3 mL/minInjection Volume:4 μLMobile Phase:Mobile Phase A: Acetonitrile:Water (60:40, v/v) with 10 mM ammonium formate and 0.1% formic acidMobile Phase B: Isopropanol:Acetonitrile (90:10, v/v) with 10 mM ammonium formate and 0.1% formic acidThe mobile phase consisted of phase A (acetonitrile/water, 6:4v/v, 0.1% formic acid) and phase B (isopropanol/acetonitrile, 9:1 v/v, 0.1% formic acid). The gradient elution program was as follows: 0.0 - 0.4 min, 30% B; 0.4 - 1.0 min, 30%-45% B; 1.0 - 3.5 min, 45%-60% B; 3.5-5.0 min, 60-75% B; 5.0-7.0 min, 75%-90% B; 7.0-8.5 min, 90-100% B; 8.5-8.6 min, 100% B; 8.6-8.61 min, 100-30% B; 8.61-10 min, 30% B.Phosphoethanolamine Cytidylyltransferase 2 Integrates DAG Metabolism and TBK1 Activation to Regulate Antiviral Innate Immunity.Ocean university of Chinazengjie LiuCell Linemass spectrometry assayCell pellets were resuspended in 100 μL of ice-cold 80% methanol, followed by addition of 300 μL ice-cold MTBE. After vigorous vortexing, samples were centrifuged at 3000 rpm for 15 min at 4°C. The upper organic phase (200 μL) was collected, vacuum-dried, and reconstituted in 200 μL of dichloromethane:methanol (1:1). The reconstituted extracts were centrifuged again (3000 rpm, 15 min, 4°C) and supernatants transferred to autosampler vials for UPLC-HRMS analysis. All steps were performed on ice.Pooled quality control (QC) samples were prepared by mixing equal aliquots of each sample extract. QC samples were injected every 10 study samples throughout the analytical run to monitor system stability and data quality.Mus musculushttps://www.ebi.ac.uk/metabolights/MTBLS14085Zengjie Liu. Ocean university of China. No. 5 Yushan Road, Shidong District, Qingdao City, Shandong Province; China University of Oceanography, Yushan Campus: 266005. 1839043781@qq.com.Qinghui Ai. Ocean University of China. qhai@ouc.edu.cn.Wencong Lai. WClai@stu.ouc.edu.cn.Step 1: Peak extraction and alignmentThe raw mass spectrometry data (.raw files) were converted to mzML format using MSConvert (ProteoWizard, version 3.0). Peak detection, alignment, and quantification were performed using XCMS (version 3.12.0) in R (version 4.1.0). Parameters were set as follows: method = 'centWave', ppm = 20, peakwidth = c(6, 17), mzdiff = 0.001, snthresh = 6. Retention time correction was performed using the 'obiwarp' method.Step 2: Quality control filteringFeatures with > 50% missing values in QC samples were removed. Data were normalized using probabilistic quotient normalization (PQN).TreatmentGenetype1839043781@qq.comRAW264.7 murine macrophage cell lines, including wild-type (Pcyt2+/+) and Pcyt2 knockout (pcyt2-/-) were used in this study. Cells were cultured in DMEM medium supplemented with 10% fetal bovine serum (FBS) and 1% penicillin-streptomycin at 37°C in a 5% CO2 humidified incubator.Three experimental groups were established, each with six biological replicates:Control group: Untreated Pcyt2+/+ RAW264.7 cells (Pcyt2+/+).VSV-infected group: Pcyt2+/+ RAW264.7 cells infected with vesicular stomatitis virus (VSV) (Pcyt2+/+ + VSV) for 12 h.Knockout group: Untreated pcyt2-/- RAW264.7 cells (pcyt2-/-).After treatment, cells were harvested by trypsinization and pelleted by centrifugation at 800 rpm for 5 minutes. The supernatant was discarded, and the cell pellets were immediately transferred to sterile centrifuge tubes, flash-frozen in liquid nitrogen, and subsequently stored at -80°C until further analysis. MetabolomicsRAW 264.7 CellsLipidomicsuntargeted metabolite profilingRAW 264.7 CellsLipidomicsuntargeted metabolite profilingInstrument: Q-Exactive spectrometer (Thermo Fisher Scientific, Bremen, Germany)Ionization Mode: Electrospray ionization (ESI) in both positive and negative ion modes (separate runs)Source Parameters (common for both modes):Sheath Gas Flow Rate: 45 (arbitrary units)Auxiliary Gas Flow Rate: 12 (arbitrary units)Sweep Gas Flow Rate: 0 (arbitrary units)Capillary Temperature: 350°CSpray Voltage: +3800 V (positive mode) / -3500 V (negative mode)Acquisition Mode: Data-dependent acquisition (DDA, also known as TopN)Full Scan (MS1) Parameters:Scan Range: m/z 70 - 1500Resolution: 70,000 (at m/z 200)AGC Target: 3e6Maximum Injection Time: 100 msMS/MS (MS2) Parameters:Isolation and fragmentation: Top 6 most abundant ions (with intensity threshold > 1e5) were selected for MS/MS fragmentationResolution: 17,500 (at m/z 200)Maximum Injection Time: 50 msDynamic Exclusion: 6 secondsQC Injection Frequency: QC samples were injected every 10 study samples throughout the batch to monitor system stability and performance.NEG_225.2218_183.7567_1NEG_747.5156_357.9582_1NEG_674.5055_369.5694_1NEG_450.2621_87.7348_1NEG_271.2065_61.76_1NEG_742.53_360.7101_1NEG_351.2173_61.807_1NEG_704.5582_411.3913_1NEG_564.5348_398.7903_1NEG_766.5345_363.7897_1NEG_317.2118_136.7875_1NEG_328.236_181.3083_1NEG_743.4856_236.7386_1NEG_465.3173_207.4381_1NEG_325.1839_117.0253_1NEG_271.2276_183.7386_1NEG_728.5528_392.4594_1NEG_646.6126_434.0462_1NEG_464.314_143.1678_1NEG_634.6128_443.4549_1NEG_483.2711_147.9746_1NEG_130.0862_57.3044_1NEG_766.5396_572.9989_1NEG_721.501_325.3615_1NEG_794.5581_377.1869_1NEG_767.4858_274.6159_1NEG_436.2826_128.2453_1NEG_283.2429_181.3083_1NEG_773.5277_310.8624_1NEG_522.283_94.9579_1NEG_464.3143_169.4133_1NEG_714.5062_325.7291_1NEG_524.2985_61.0656_1NEG_756.5533_391.2112_1NEG_476.2775_96.4374_1NEG_647.4649_447.5757_1NEG_307.2641_251.0221_1NEG_327.2327_181.299_1NEG_253.2162_303.6428_1NEG_747.5177_284.9313_1NEG_774.5427_381.8116_1NEG_319.2275_157.7502_1NEG_538.5195_386.525_1NEG_151.0251_57.313_1NEG_536.5034_377.7296_1NEG_592.5657_420.0493_1NEG_702.5428_409.3275_1NEG_746.5115_358.477_1NEG_297.2431_188.4839_1NEG_828.6482_446.8232_1NEG_690.506_344.1782_1NEG_722.5122_404.9179_1NEG_478.293_120.0123_1NEG_785.6521_423.778_1NEG_478.2931_153.4519_1NEG_275.1651_548.5689_1NEG_253.217_195.6254_1NEG_299.2589_231.4671_1NEG_718.5372_367.1535_1NEG_480.3089_144.7546_1NEG_281.2482_239.294_1NEG_718.5285_379.4879_1NEG_1403.9904_482.8423_1NEG_717.4701_274.6489_1NEG_772.5271_359.0234_1NEG_407.2945_301.4157_1NEG_279.1963_95.5033_1NEG_744.5533_398.4981_1NEG_771.6384_435.8234_1NEG_331.2641_228.2528_1NEG_771.6347_415.0299_1NEG_799.6596_412.8375_1NEG_716.522_379.5238_1NEG_676.5272_392.7704_1NEG_770.5684_402.3278_1NEG_329.2484_209.6211_1NEG_797.6503_412.1242_1NEG_303.2326_191.8105_1NEG_409.3102_311.4616_1NEG_409.301_300.6449_1NEG_279.2326_204.4965_1NEG_509.2876_100.8197_1NEG_275.1652_572.6922_1NEG_275.1651_515.5564_1falsePhosphoethanolamine Cytidylyltransferase 2 Integrates DAG Metabolism and TBK1 Activation to Regulate Antiviral Innate ImmunityPhosphatidylethanolamine (PE) biosynthesis is critical for membrane biology and cellular homeostasis. However, its specific role in antiviral innate immunity remains poorly understood. Here, we demonstrate that inhibition of phosphoethanolamine cytidylyltransferase 2 (PCYT2), a key enzyme in PE biosynthesis, promotes TBK1 activation to enhance antiviral immune response. Mechanistically, PCYT2 deficiency leads to the accumulation of diacylglycerol (DAG), which activates protein kinase C δ (PKCδ). We identify PKCδ as a direct kinase for TBK1, and demonstrate that it binds to and phosphorylates TBK1 at Ser716. This Ser716 phosphorylation facilitates subsequent canonical phosphorylation at Ser172, resulting in hyperactivation of the TBK1–IRF3 axis. Our findings uncover a novel link between phospholipid metabolism and antiviral innate immunity, suggesting that targeting the PE biosynthesis pathway could be a potential therapeutic strategy against viral infections.2026-05-072026-03-19MTBLS14085LMFA01010001LMFA01010012LMFA01010014LMSP03010029LMGP01010481LMFA07070060LMFA08040007LMGP01010560LMGP01010884LMGP01010768LMSP03010002LMGP01010882LMGP01010739LMGP01010005LMGP01010488LMGP01010684LMFA08010009LMGP01010594LMGP01010477LMGP01010530LMGP01010937LMFA07070098LMGP01010956LMFA08040038LMGP01010802LMFA07070079LMGP01050012LMGP01030013LMGP01050024LMGP01010564LMFA07010474LMFA08040004LMGP01050018LMGP01010485LMGP01070009LMGP01050016LMSP03010001LMSP03010003LMGP01050045LMFA07070107LMFA08040051LMFA08040015LMGP01050113LMGP01011014LMGP02010337LMST01010059LMGP01011052LMGP01050026LMGP01010532LMGP01011110LMGP01010573LMFA01010019LMGP01060014LMGP01010456LMPR0102010015LMFA08010020LMGP01010707LMFA07070099LMSP02010012LMGP01050121LMGP02050002LMFA07070004LMGP01010006LMPR02010001LMGP01050114LMGP01060010LMGP01020256LMGP01050057LMGP01050082LMSP03010077LMGP01050032LMGL03010002LMSP02010008LMFA08020200LMST01020007LMST01020012LMFA08010008LMGP01050076LMSP02010004LMST01020025LMGP02011250LMPR0102090029LMGP02050001LMFA08020207LMFA07070008LMGP01070006LMGP02010425LMGP01010976LMFA01031043LMGP01030030LMFA07010891LMGP02011228LMGP03010040LMSP03010081LMFA08010004LMGP02010096LMGL03010250LMGP02010095LMGP02030003LMGP03010916LMGP03010771LMFA08040013LMGP12010118LMFA08040009LMGP01050022LMGP01050035LMFA06000255LMGP02010111LMGP01020046LMST01020005LMGL03010001LMFA08010001LMFA01010004LMGP01020012LMGP01020261LMST01020004LMGP01050056LMST04010123LMPR0103140001LMFA01030056LMST04010001LMFA01030073LMFA01030120LMST02020079LMFA03080014LMFA01030001LMPR02020057LMFA01030178LMPR02020055LMFA06000083LMFA04000064LMFA01030185LMFA01030377LMGP02050004LMGP02020094LMFA01030057LMFA03070010LMGP02010326LMGP04010967LMFA05000652LMGP02011220LMGP02010036LMGP04010982LMGP02010009LMFA05000643LMGP04010008LMGP02030040LMGP02010052LMFA02000345LMFA03010087LMGP02010325LMFA01050051LMFA02000130LMGP04010901LMGP04050006LMGP02010118LMFA05000664LMGP04010986LMGP04050008LMSP02020001LMFA01100035LMGP02011200LMGP02030093LMGP10010012LMSP02010009LMGP02010338LMFA07070046LMFA05000662LMSP02010006LMGP03050001LMSP02010007LMSP02010021LMGP02030001LMGP02030013LMSP03010075LMGP02050010LMGP02050038LMGP04010899LMGP02070002LMFA05000687LMGP02011225LMGP02070001LMGP02030006LMSP03010078LMGP02050011LMST05030001LMGP03050006LMSP03010067LMGP02050064LMFA07010960LMGP02010037LMSP03010065LMST05020016LMFA03050002LMSP03010006LMGP02030005LMGP02010294LMGP02020103LMGP04010987LMGP02030004LMGP02010943LMGP12010116