#METABOLOMICS WORKBENCH juliehaines_20221216_071050 DATATRACK_ID:3653 STUDY_ID:ST002413 ANALYSIS_ID:AN003933 VERSION 1 CREATED_ON 12-19-2022 #PROJECT PR:PROJECT_TITLE IFN-inducible phospholipid levels govern endosomal antiviral immunity PR:PROJECT_SUMMARY The interferon-induced transmembrane proteins (IFITMs) are implicated in several PR:PROJECT_SUMMARY biological processes including antiviral defense, but their modes of action PR:PROJECT_SUMMARY remain debated. Here, taking advantage of pseudotyped viral entry assays and PR:PROJECT_SUMMARY replicating viruses, we uncover the requirement of host co-factors for endosomal PR:PROJECT_SUMMARY antiviral inhibition through high-throughput proteomics and lipidomics in PR:PROJECT_SUMMARY cellular models of IFITM restriction. Unlike plasma membrane (PM)-localized PR:PROJECT_SUMMARY IFITM restriction that targets infectious SARS-CoV2 and other PM-fusing viral PR:PROJECT_SUMMARY envelopes, inhibition of endosomal viral entry depends on lysines within the PR:PROJECT_SUMMARY conserved IFITM intracellular loop. These residues recruit PR:PROJECT_SUMMARY phosphatydilinositol-3-phosphate (PIP3) that we show here to be required for PR:PROJECT_SUMMARY endosomal IFITM activity. We identify PIP3 as an IFN-inducible phospholipid that PR:PROJECT_SUMMARY acts as a rheostat for endosomal antiviral immunity as its expression levels PR:PROJECT_SUMMARY correlated with the potency of endosomal IFITM restriction and exogenous PIP3 PR:PROJECT_SUMMARY enhanced inhibition of endocytic viruses including the recent SARS-CoV2 Omicron PR:PROJECT_SUMMARY variant. Together, our results identify PIP3 levels as a critical regulator of PR:PROJECT_SUMMARY endosomal IFITM restriction linking it to the Pi3K/Akt/mTORC pathway and PR:PROJECT_SUMMARY elucidate cell-compartment specific antiviral mechanisms, informing the PR:PROJECT_SUMMARY development of broadly acting antiviral strategies. PR:INSTITUTE University of Colorado Denver PR:LABORATORY Angelo D'Alessandro PR:LAST_NAME Haines PR:FIRST_NAME Julie PR:ADDRESS 12801 E 17th Ave, Room 1303, Aurora, Colorado, 80045, USA PR:EMAIL julie.haines@cuanschutz.edu PR:PHONE 3037243339 PR:DOI http://dx.doi.org/10.21228/M8ZB01 #STUDY ST:STUDY_TITLE IFN-inducible phospholipid levels govern endosomal antiviral immunity ST:STUDY_SUMMARY Untargeted lipidomics of 3 cell lines at baseline with focus on phospholipids to ST:STUDY_SUMMARY understand the role of these lipids in endosomal antiviral immunity ST:INSTITUTE University of Colorado Denver ST:LABORATORY Angelo D'Alessandro ST:LAST_NAME Haines ST:FIRST_NAME Julie ST:ADDRESS 12801 E 17th Ave, Room 1303, Aurora, Colorado, 80045, USA ST:EMAIL julie.haines@cuanschutz.edu ST:PHONE 3037243339 ST:SUBMIT_DATE 2022-12-16 #SUBJECT SU:SUBJECT_TYPE Cultured cells SU:SUBJECT_SPECIES Homo sapiens #SUBJECT_SAMPLE_FACTORS: SUBJECT(optional)[tab]SAMPLE[tab]FACTORS(NAME:VALUE pairs separated by |)[tab]Additional sample data SUBJECT_SAMPLE_FACTORS - F2-27-07 Group:K562 Oe-IFITM3 RAW_FILE_NAME=F2-27-07 SUBJECT_SAMPLE_FACTORS - F2-27-08 Group:K562 Oe-IFITM3 RAW_FILE_NAME=F2-27-08 SUBJECT_SAMPLE_FACTORS - F2-27-09 Group:K562 Oe-IFITM3 RAW_FILE_NAME=F2-27-09 SUBJECT_SAMPLE_FACTORS - F2-27-13 Group:mPB-CD34+ RAW_FILE_NAME=F2-27-13 SUBJECT_SAMPLE_FACTORS - F2-27-14 Group:mPB-CD34+ RAW_FILE_NAME=F2-27-14 SUBJECT_SAMPLE_FACTORS - F2-27-15 Group:mPB-CD34+ RAW_FILE_NAME=F2-27-15 SUBJECT_SAMPLE_FACTORS - F2-27-01 Group:THP1 Oe-IFITM3 RAW_FILE_NAME=F2-27-01 SUBJECT_SAMPLE_FACTORS - F2-27-02 Group:THP1 Oe-IFITM3 RAW_FILE_NAME=F2-27-02 SUBJECT_SAMPLE_FACTORS - F2-27-03 Group:THP1 Oe-IFITM3 RAW_FILE_NAME=F2-27-03 #COLLECTION CO:COLLECTION_SUMMARY The human embryonic kidney 293T (HEK293T) and K562 cells were maintained in CO:COLLECTION_SUMMARY Iscove's modified Dulbecco’s medium (IMDM; Sigma), human THP1 cells were CO:COLLECTION_SUMMARY maintained in Roswell Park Memorial Institute medium (RPMI; Lonza). All media CO:COLLECTION_SUMMARY were supplemented with 10% fetal bovine serum (FBS; Gibco), penicillin (100 CO:COLLECTION_SUMMARY IU/ml), streptomycin (100 µg/ml) and 2% glutamine. A549 and Vero E6 cells were CO:COLLECTION_SUMMARY cultured in Dulbecco's Modified Eagle Medium (DMEM, Sigma) supplemented with 10% CO:COLLECTION_SUMMARY fetal bovine serum (FBS; Gibco), 1% Non-essential amino acid (NEAA, Gibco), 1% CO:COLLECTION_SUMMARY Sodium Pyruvate (Gibco). Calu3 were cultured in Dulbecco's Modified Eagle Medium CO:COLLECTION_SUMMARY (DMEM, Sigma) supplemented with 20% fetal bovine serum (FBS; Gibco), 1% CO:COLLECTION_SUMMARY Non-essential amino acid (NEAA, Gibco), 1% Sodium Pyruvate (Gibco) and 2% CO:COLLECTION_SUMMARY GlutaMAX (Gibco). All cells were maintained in a 5% CO2 humidified atmosphere at CO:COLLECTION_SUMMARY 37°C. A549-hACE2 cell lines were generated by lentiviral transduction. CO:COLLECTION_SUMMARY Lentiviral vectors were produced following a standard procedure based on calcium CO:COLLECTION_SUMMARY phosphate co-transfection with 3rd generation helper and transfer plasmids. CO:COLLECTION_SUMMARY Primary cells Human CD34+ HSPC were isolated through positive magnetic bead CO:COLLECTION_SUMMARY selection according to manufacturer’s instructions (Milteney) from umbilical CO:COLLECTION_SUMMARY cord blood or from mononuclear cells collected upon informed consent from CO:COLLECTION_SUMMARY healthy volunteers according to the Institutional Ethical Committee approved CO:COLLECTION_SUMMARY protocol (TIGET01). Otherwise, CB, bone marrow (BM) or G-CSF mPB-CD34+ cells CO:COLLECTION_SUMMARY were directly purchased from Lonza or Hemacare. All cells were maintained in a CO:COLLECTION_SUMMARY 5% CO2 humidified atmosphere at 37°C. CO:SAMPLE_TYPE Cultured cells #TREATMENT TR:TREATMENT_SUMMARY Transduction All cells were transduced at the indicated multiplicity of TR:TREATMENT_SUMMARY infection (MOI) as calculated by titration of vector stocks on 293T cells and TR:TREATMENT_SUMMARY expressed as transducing units (TU)/293T cell. Cell lines were transduced with TR:TREATMENT_SUMMARY LV for 16 hours, washed and kept in culture until reading transduction TR:TREATMENT_SUMMARY efficiency by flow cytometry. Human CB-derived HSPC were cultured in serum-free TR:TREATMENT_SUMMARY StemSpan medium (StemCell Technologies) supplemented with penicillin (100 TR:TREATMENT_SUMMARY IU/ml), streptomycin (100 µg/ml), 100 ng/ml recombinant human stem cell factor TR:TREATMENT_SUMMARY (rhSCF), 20 ng/ml recombinant human thrombopoietin (rhTPO), 100 ng/ml TR:TREATMENT_SUMMARY recombinant human Flt3 ligand (rhFlt3), and 20 ng/ml recombinant human IL6 TR:TREATMENT_SUMMARY (rhIL6) (all from Peprotech) 16 to 24 hours prior to transduction. HSPC were TR:TREATMENT_SUMMARY then transduced at a concentration of 1×106 cells per milliliter with TR:TREATMENT_SUMMARY VSV-gp-pseudotyped SINLV for 16 hours at the indicated multiplicity of infection TR:TREATMENT_SUMMARY (MOI) in the same medium. G-CSF mobilized peripheral blood CD34+ cells were TR:TREATMENT_SUMMARY maintained in culture in CellGro medium (Cell Genix) containing a cocktail of TR:TREATMENT_SUMMARY cytokines: 60 ng/ml IL-3, 100 ng/ml TPO, 300 ng/ml SCF, and 300 ng/ml FLT-3L TR:TREATMENT_SUMMARY (all from Cell Peprotech). Transductions were performed with the indicated dose TR:TREATMENT_SUMMARY of vectors for 16 hours in the same cytokine-containing medium. For single hit TR:TREATMENT_SUMMARY reporter LV transductions cells were washed and maintained in serum-free medium TR:TREATMENT_SUMMARY supplemented with cytokines as above until the reading of the percentage of TR:TREATMENT_SUMMARY positive cells by FACS. To perform OE, KD and KO experiments cells were TR:TREATMENT_SUMMARY transduced with the respective LV and then challenged with a second transduction TR:TREATMENT_SUMMARY with or without drugs. Except for primary cells, KD and KO cells were selected TR:TREATMENT_SUMMARY with Puromycin before the second hit of transduction. #SAMPLEPREP SP:SAMPLEPREP_SUMMARY Lipids were extracted from frozen cell pellets using LC-MS grade cold methanol SP:SAMPLEPREP_SUMMARY at a concentration of 2e6 cells/mL. Samples were briefly vortexed to mix then SP:SAMPLEPREP_SUMMARY incubated for 30 min at -20C. Supernatants were clarified by centrifugation (10 SP:SAMPLEPREP_SUMMARY min, 18,213 g, 4C) then 25 uL aliquots were transferred to autosampler vials. #CHROMATOGRAPHY CH:CHROMATOGRAPHY_SUMMARY 17 minute negative CH:INSTRUMENT_NAME Thermo Vanquish CH:COLUMN_NAME Waters ACQUITY UPLC HSS T3 (150 x 2.1mm,1.8um) CH:COLUMN_TEMPERATURE 45 CH:FLOW_GRADIENT 0-1 min 25% B and 0.3 ml/min; 1-2 min 25-50% B and 0.3 mL/min; 2–8 min CH:FLOW_GRADIENT 50–90% B and 0.3 mL/min; 8–10 min 90–99% B and 0.3 mL/min; 10–14 min CH:FLOW_GRADIENT hold at 99% B and 0.3 mL/min; 14–14.1 min 99–25% B and 0.3 mL/min; CH:FLOW_GRADIENT 14.1–16.9 min hold at 25% B and 0.4 mL/min; 16.9–17 min hold at 25% B and CH:FLOW_GRADIENT resume flow of 0.3 mL/min CH:FLOW_RATE varies, see solvent gradient CH:SOLVENT_A 25% acetonitrile; 5 mM ammonium acetate CH:SOLVENT_B 50% isopropanol/45% acetonitrile/5% water; 5 mM ammonium acetate CH:CHROMATOGRAPHY_TYPE Reversed phase #ANALYSIS AN:ANALYSIS_TYPE MS #MS MS:INSTRUMENT_NAME Thermo Q Exactive Orbitrap MS:INSTRUMENT_TYPE Orbitrap MS:MS_TYPE ESI MS:MS_COMMENTS the mass spectrometer operated in top 10 data dependent MS2 mode with NCE of 30 MS:MS_COMMENTS eV (HCD) and dynamic exclusion of 10 s. MS1 scans were acquired at a resolution MS:MS_COMMENTS of 70,000 across the m/z range of 125-1500. MS2 scans were acquired at a MS:MS_COMMENTS resolution of 17,500. Sheath and auxiliary gases (both N2) were set at 45 and 15 MS:MS_COMMENTS (unitless), respectively. Instrument stability and quality control were assessed MS:MS_COMMENTS via injection of technical replicates every 6 runs. Untargeted lipid results MS:MS_COMMENTS were obtained using LipidSearch (Thermo) with precursor ion tolerance set to 5 MS:MS_COMMENTS ppm and product ion tolerance at 8 ppm. MS:ION_MODE NEGATIVE #MS_METABOLITE_DATA MS_METABOLITE_DATA:UNITS peak area MS_METABOLITE_DATA_START Samples F2-27-07 F2-27-08 F2-27-09 F2-27-13 F2-27-14 F2-27-15 F2-27-01 F2-27-02 F2-27-03 Factors Group:K562 Oe-IFITM3 Group:K562 Oe-IFITM3 Group:K562 Oe-IFITM3 Group:mPB-CD34+ Group:mPB-CD34+ Group:mPB-CD34+ Group:THP1 Oe-IFITM3 Group:THP1 Oe-IFITM3 Group:THP1 Oe-IFITM3 LPE(16:0) 54997.8546 56008.7811 40932.2682 84105.5155 94440.7059 125245.8385 808422.1281 270338.6235 902172.3621 LPE(18:0) 51994.8749 78004.2275 71756.5024 186196.8685 123011.7149 169845.9397 3402227.0240 1691963.8900 4979101.3260 LPE(18:1) 121276.8196 247196.5067 85035.4018 330228.9067 345873.7716 568844.4624 738576.6416 742456.9987 1152855.2050 LPE(18:2) 50920.6819 174967.8120 45826.0353 58225.1918 149157.6833 318300.4518 185314.6779 448048.5437 304172.8850 LPE(20:4) 171499.1337 215537.1088 109974.1002 308638.9094 337391.8897 384163.2723 155385.3099 285531.9697 458548.7268 LPE(20:5) 27214.7927 174549.3865 67137.1189 16762.1415 110915.9967 222674.3807 202670.4285 395734.6350 362772.4076 LPG(18:1) 15376.6941 7773.2879 10652.6813 31630.9229 19883.4897 6932.8270 385317.5140 371030.6634 1315655.7440 LPG(20:0) 6020.3462 27098.0705 22137.3478 12408.8402 5589.3435 9881.3779 287299.9135 264215.2199 1023020.4880 LPI(18:0) 99745.3908 219252.0377 99895.0262 89891.0054 89982.8905 130382.3838 1499566.3430 875209.4396 2212562.3740 PC(14:0_18:3) 69343.8817 54635.9109 51446.3147 95821.7859 51390.0056 53104.5029 2707190.9530 1941941.5180 5702871.6360 PC(14:0_20:5) 83166.7316 113471.6394 52530.5996 75726.9416 114343.9768 81218.4775 1765838.7770 2088714.3180 7176207.8300 PC(14:1e_18:2) 75596.9974 74569.4903 45659.0318 47148.6424 45954.2149 47503.5606 9116438.1120 6370723.9370 18768013.8400 PC(14:1e_18:4) 119634.5682 43086.2862 63237.6868 41857.0411 62013.5792 69161.8976 3010905.8830 3635443.5850 9048391.6480 PC(15:0_16:1) 2190433.3550 1464807.3260 2365065.7440 10745693.4700 2615703.5710 8116948.9950 150025497.0000 70142985.7500 223853436.2000 PC(15:0_18:3) 1027203.3550 751699.4250 1588285.6720 10819031.5200 1865533.3050 7517447.3020 64237867.7500 49660649.5000 206774840.1000 PC(16:0_16:1) 209446.4804 124796.9975 400899.7702 404188.9875 36646.3632 240141.3849 14958198.7800 9878108.6380 31334934.6100 PC(16:0_18:1) 57727.3061 25677.0108 134787.1063 40609.7400 14086.7343 46574.6116 3546760.1890 2105076.5490 7695383.2780 PC(16:0_18:3) 41312.3405 45561.5190 109895.7407 360808.9166 145569.6459 163162.9635 3685027.8690 2643026.5430 5472997.2790 PC(16:0e_20:5) 55944.7980 125050.0312 65475.1049 179443.1922 59671.4509 294020.6597 16686965.4900 10032657.5300 19519670.2900 PC(16:1_14:0) 68133.7583 57316.0201 123965.8618 82664.1366 40835.7097 41940.2881 8387379.5940 4519257.0430 12924443.8500 PC(16:1_16:1) 77930.5102 48030.4719 153096.7815 310305.7240 77046.0743 234501.1425 8602347.0350 7976127.5160 23870281.4500 PC(16:1_18:1) 59377.1411 63706.2620 75788.3094 230980.4662 60892.6043 108880.4945 3131882.9190 1799547.2170 6965097.6790 PC(16:1e_18:3) 53124.2985 58005.8634 148030.3091 68888.0439 55829.9198 71218.0941 6787541.5740 3661220.1760 11868439.3500 PC(17:0_18:1) 25804.4406 22220.2289 21133.1708 47783.0815 21844.6868 27484.8622 1748428.2960 1094569.9490 3521519.4950 PC(17:1_16:0) 2596651.6570 2111318.4730 3942375.2490 16754405.8300 2893074.3780 12615453.0900 156564444.0000 97568971.7300 303112718.8000 PC(17:1_16:1) 3787246.7920 3558700.9640 4969425.6170 28345513.9600 5129120.8700 22008419.5100 218918275.7000 159043586.3000 401859736.4000 PC(17:1_18:2) 190216.3877 119468.9987 404611.8167 5261044.3000 849981.6616 3735743.4540 25147175.1100 23644921.1900 82767559.7200 PC(17:1_18:3) 2268792.6730 1361828.5680 2979791.5440 54444709.1700 9013412.0610 40810785.7300 90251892.1800 66128898.7600 183543861.4000 PC(18:0_18:1) 16977.9897 30089.9602 21015.0686 11085.6868 10846.3061 17441.4444 930983.0300 536199.1903 1533689.6590 PC(18:1_18:2) 147512.3631 137895.2493 143406.1085 193324.1682 233574.3424 261260.1123 2619843.6930 995675.8599 3228142.9080 PC(18:1_20:3) 41431.2792 140885.9580 43648.7567 62226.2314 42305.3624 80072.1871 2589572.1130 1160191.5700 5524341.1080 PC(18:1e_22:5) 108657.9127 4207155.5070 425781.3164 15350718.6400 288778.0266 12177454.4800 222977946.1000 159764435.4000 78268672.9700 PC(18:1e_22:6) 673432.9585 542063.7868 55157.9793 10247299.0200 1928705.4940 6665093.3370 75452734.4400 78250624.7700 142328489.8000 PC(32:4e) 49322.2524 56193.7028 52602.4793 53952.6975 52141.1140 54546.3608 4259949.2050 4149962.8440 10806422.4400 PC(34:6e) 31461.7574 61010.0839 222090.7699 59606.9839 93357.1948 87992.4083 15864167.5900 15313197.0000 41659038.1500 PC(36:6e) 2028337.9760 1337650.1610 3399948.5480 50748371.1400 8429992.8180 37646172.4700 73792767.4000 5480629.0370 186464756.3000 PC(36:7e) 25457.8600 93073.8685 26544.7582 27527.5438 26608.9947 168165.2697 2309149.8770 2264974.7410 6399320.2200 PE(16:0_16:1) 64183.9939 85474.3164 39678.0199 1286611.6620 175856.8194 835379.3005 42725190.1100 12522370.3200 40046160.0600 PE(16:0_18:3) 86284.4751 123467.0621 89963.7059 776065.2419 95448.5395 772481.0416 17405591.0200 7730944.1430 27027469.8300 PE(16:1_16:1) 41962.0853 48037.5757 43347.8598 210205.6588 43917.5247 98909.5207 3808215.3920 1011705.3200 4262734.1460 PE(16:1_18:1) 382219.7707 178611.7770 526982.6610 5303195.6880 1055346.1320 3980301.8480 68361087.4800 32374078.8200 101161433.1000 PE(16:1e_16:1) 58072.9962 38849.9818 72070.7089 59141.2460 37249.2554 38653.1186 51862502.8500 10633306.1400 34046103.2700 PE(16:1e_18:1) 1410310.6320 1070366.1400 2013289.6780 1406431.2690 236622.3204 755187.0622 178717489.3000 62334912.2900 223332419.0000 PE(16:1e_18:2) 206837.3219 484711.9327 482852.8446 394956.6229 37549.7304 524287.0976 118891497.9000 58175062.1300 168270548.8000 PE(16:1e_18:3) 40704.8867 52651.0373 50243.2756 49580.4917 36351.1992 47075.0928 29874055.6000 11012401.6800 30716124.7000 PE(16:1e_20:3) 219806.6306 601196.2443 621468.8844 703965.8298 119042.1813 380659.9762 102082201.5000 47612346.5000 171221418.3000 PE(16:1e_20:4) 783698.3342 639202.3732 1356609.2340 17396634.5900 2871467.3900 12266197.8100 141083020.7000 92451461.2400 293067466.5000 PE(16:1e_20:5) 439567.3895 361809.0651 701088.8915 227643.7836 204980.1643 493251.7195 242513007.9000 130980914.7000 481165786.7000 PE(16:1e_22:5) 660053.9524 1900258.5340 1173633.9910 243963.5966 426448.0790 508945.1073 179602551.7000 139475322.3000 235008365.3000 PE(18:0_14:0) 14866.2123 16805.0632 15564.9661 52928.6617 15264.0906 17300.7366 23015740.5100 8247347.8970 13459645.8400 PE(18:0_22:4) 50581.6764 80682.0692 104518.4195 465586.0613 65014.2815 330738.7482 5133981.7200 2613013.6990 11282542.7700 PE(18:1_20:4) 952323.2400 426280.6186 2101648.4960 3916697.2620 235721.8671 1501772.6960 80221485.0700 57316818.6600 221791684.6000 PE(18:1_20:5) 267297.3891 199836.6993 450141.0256 2757200.4740 417305.3358 1888293.3560 26633506.5300 21401203.2100 72708091.9700 PE(18:1_22:4) 1265917.1880 574734.9253 2448672.3420 5121159.8400 1329865.7020 3533511.9800 43964255.5300 45038042.1200 36061095.0600 PE(18:1_22:5) 111981.5297 163535.5576 161612.6224 4290192.0400 832629.5970 3128282.8950 11223948.4200 12693136.1800 45062724.6100 PE(18:1_22:6) 21224.4704 23533.3704 33533.2782 1875303.5770 134089.3209 720468.8410 3361650.9600 3188302.8940 12891201.9200 PE(18:1e_18:2) 1006974.3440 2476967.6510 1415623.2330 819719.6803 407035.2088 965764.2039 94343990.3800 49056443.7200 173551151.7000 PE(18:1e_20:3) 155685.8050 116350.6994 203783.0776 524643.0393 105883.0803 503543.8195 28721904.2300 22254590.9000 73823198.8100 PE(18:1e_20:4) 1530858.9180 737015.7505 2340494.0800 18544991.7900 2793369.4310 11288540.7200 71602455.1200 59197323.9400 203709022.3000 PE(18:1e_22:4) 63003.3158 52316.0269 44045.4545 2082030.2430 219251.4762 1398464.5910 3110540.7650 3191394.0000 14091678.6200 PE(18:1e_22:5) 857427.3388 671842.1874 1418642.1850 3237795.1170 550177.4953 2168951.4000 47420154.5300 49888201.6000 179918829.5000 PE(18:2_18:2) 90254.8818 55732.9151 175001.6024 1493135.2890 260307.5198 1125527.8170 14464792.7600 12436271.0200 37879204.7200 PE(18:2e_22:5) 201001.5522 167336.9247 321174.1937 1430068.8510 170368.1439 787580.9008 29622367.9300 25961517.0900 81334787.6400 PE(18:2e_22:6) 34722.0880 66400.8660 64081.7145 449997.7067 51521.8293 253918.9987 6462134.3730 5237412.1100 11690205.2400 PE(18:3_18:2) 41776.5540 54301.6706 51727.4241 27066.6798 130197.0457 813563.9519 15249429.8700 8607441.9360 31371992.3600 PE(20:5_18:2) 138186.8417 37080.5448 148294.1061 383546.8132 323343.5169 150601.7773 2088713.6260 999115.6470 4427181.1530 PE(33:2e) 40952.2945 90389.5927 75738.4451 56114.2609 43618.1279 45088.7125 10572856.3600 4039648.6250 15485413.4600 PE(34:6e) 24600.7703 34258.2833 32842.8853 24045.9694 25676.8424 28339.1817 3398654.8770 1415368.5850 4900397.6410 PE(38:7e) 376563.8263 310957.5293 556643.9739 879259.7549 128893.4998 767646.4225 73551480.9800 35433263.7200 175209802.2000 PG(15:0_18:1) 7803.2192 16079.7222 8462.8464 7852.0704 8875.6339 12359.1748 929118.1156 574132.4496 2313463.4830 PG(16:0_16:0) 16049.5400 50613.2818 19459.9767 77257.5824 46861.6948 55582.7306 1394556.9390 401400.4758 1576594.3660 PG(16:0_18:1) 615326.9834 392046.3436 598909.4050 2242070.8290 662532.8630 1712449.9420 56545188.8600 41308611.5400 142913243.4000 PG(16:1_18:1) 14977.6686 16741.2693 15690.0893 166843.7190 54440.4929 176652.2905 4220811.4270 2058863.2610 7944339.2390 PG(17:0_18:1) 50938.2238 72459.4605 65071.9213 72995.4979 208511.4789 471440.5229 9566598.8480 4152508.7170 17223304.7900 PG(17:1_18:1) 48323.9546 71729.5593 24271.8164 23377.0900 25725.3633 216600.6106 1288875.3790 724489.5493 2254821.0870 PG(18:1_14:0) 10582.6328 96495.8412 10861.9264 58420.5964 17856.3043 136943.1486 2243106.4870 732938.6382 2571758.5430 PG(18:1_18:1) 183342.4695 351675.7441 228269.6027 883993.5241 158821.3922 370407.4764 5248698.3480 5127956.3030 18702470.3900 PG(18:1_18:2) 23131.4075 17758.0838 29979.8246 147576.7853 36687.3068 49087.0932 1948165.1600 2273860.2670 7462547.1570 PG(18:1_18:3) 22528.2923 25133.6635 28194.2209 40652.4379 23598.8207 36920.1027 859944.8444 618293.9104 2090621.3560 PG(20:0_16:0) 5751.0194 48438.4673 6727.4811 6067.6825 10626.1055 23130.2009 477715.7655 200375.1255 1188527.8790 PG(20:0_18:1) 124811.7897 992402.6831 57268.2273 47036.9716 418866.7907 1015374.9980 6120321.2730 2393022.6830 12288456.0400 PG(20:0_18:2) 22427.4859 214965.3258 19888.7583 19713.9971 69684.0863 228650.9665 1530556.2560 1011215.9950 3466830.0410 PG(20:0_20:3) 11638.4270 101311.2600 6529.9911 6491.7375 8195.1797 96931.2156 1017517.6690 1168082.8000 8791377.1450 PG(20:0_22:5) 5856.6921 18040.8728 5971.6098 5600.6235 6273.5508 20827.1387 165654.6101 341898.3938 1782074.0470 PG(20:0_22:6) 6437.6723 22733.0310 6607.5733 6447.9255 6488.3243 30527.2295 981463.2011 1621779.4320 6462361.8550 PI(15:0_18:1) 19162.0907 26830.7702 19701.0526 31820.5508 20525.9809 21420.8008 2299615.9940 624421.3596 3002289.1900 PI(16:0_16:0) 21506.8210 336339.1418 20570.1231 52053.1926 155049.3400 404075.3150 2661665.4120 129606.2459 1956105.7960 PI(16:0_16:1) 25968.8558 142236.4051 14565.6472 160645.2599 59817.2860 169328.3028 8116272.2660 1435610.1930 5878587.6450 PI(16:0_18:1) 370448.0371 279134.3779 709336.7341 2310940.5670 425173.3611 1881424.1990 93446810.5600 25408985.6400 85341751.8800 PI(16:0_18:3) 19322.4368 27534.4212 21308.8629 33916.6442 19466.2286 41586.8025 1450202.1430 721897.5997 2790927.5930 PI(16:0_20:5) 10044.0333 14401.2294 9494.5127 8618.9308 9752.3304 12801.1177 3247724.7610 2232872.8820 8769886.0630 PI(16:0e_20:3) 16850.0894 19385.5988 17557.2307 17989.6285 17894.0904 21661.9340 1439580.3330 648826.8143 2426241.8820 PI(16:0e_20:5) 8506.4043 9890.8697 8704.6117 8487.4516 8885.6685 9455.4059 678377.1879 387062.8475 1353251.7880 PI(16:0e_22:5) 13471.4003 20784.9472 13850.3032 63613.6139 22029.0130 47448.7812 5105392.3760 3010052.4910 9439560.3020 PI(16:1_18:1) 72808.1477 52145.8434 90160.9333 1557466.5460 330799.3401 1294871.6250 22233324.6200 7238852.6790 27274586.2000 PI(17:0_18:1) 41682.0665 66702.5096 191567.5339 179914.4387 38450.5228 101056.0960 10796522.8300 3559851.7070 12534686.6800 PI(17:0_20:3) 32096.4295 19386.5121 31389.9738 23560.7088 18943.9184 19386.8354 5514224.6460 2842614.5170 11187626.0700 PI(17:0_20:4) 34824.8235 21052.4900 32889.9638 196441.2635 28438.6091 75187.6561 2452334.9220 2075729.7260 8308422.3470 PI(17:0_20:5) 10660.7639 12566.5647 10899.3423 10761.2599 10855.0277 11926.7756 673211.7259 689169.8062 3064805.2330 PI(18:0_18:1) 2681813.4180 1689167.3480 3060282.9560 5211628.1410 1210237.0840 5414011.5970 196168533.3000 61159070.5400 178741730.1000 PI(18:0_20:1) 104448.0416 47694.3407 94604.5623 244385.7386 43661.8718 151705.1248 5006742.5860 1545779.0940 6194240.6400 PI(18:0_20:2) 1018625.8080 439682.2767 1736187.0700 451069.1590 48268.2754 973250.0055 51576673.9700 17149908.7300 61479395.2100 PI(18:0_20:3) 142349.0799 413971.5516 356341.0848 4712508.9320 664691.8462 3421564.8200 136520556.5000 55394205.3700 285305068.1000 PI(18:0_20:4) 3237916.9890 2005952.1720 6294534.8440 45434031.2400 7467261.3920 30887970.2700 161090728.0000 147752462.9000 523079421.1000 PI(18:0_22:2) 29107.0736 131670.6512 48708.6188 383781.5837 49918.0953 360489.7607 2281599.0940 620949.6884 1916281.7450 PI(18:0_22:3) 72436.8322 106814.6060 31356.5429 267868.7207 44535.9743 368834.2324 5400059.5400 2564708.1080 9217827.7670 PI(18:0_22:4) 158368.4084 62888.0249 431964.5089 5987389.8390 710269.4412 3770841.7710 11950019.1200 9115343.3890 31345472.8500 PI(18:0_22:5) 410856.8172 248844.4304 811609.7502 4642051.8250 893709.2213 3398584.2610 33766542.7700 24712187.0500 89497211.6800 PI(18:0e_20:4) 69273.0838 22681.0535 37175.6865 62829.7237 30104.1834 126300.2118 6147283.9400 8435671.9320 25505289.5400 PI(18:0e_22:5) 15539.9885 57610.0159 15927.4403 15441.4703 17019.2632 20064.1629 5022042.6330 3726944.0900 13220427.9000 PI(18:1_18:1) 1327997.5140 988395.7573 2391364.6710 10582050.6900 1679550.5160 8085822.3570 160377672.8000 65853022.3000 238294004.9000 PI(18:1_18:2) 141665.7831 204457.9764 347615.8536 1864337.5420 490927.5954 1898974.3290 25030823.2400 15570756.4400 58209777.1900 PI(18:1_18:3) 39315.8499 57233.7832 46283.7432 1207382.0550 212674.6797 906593.6642 8911212.9850 6545533.6930 27867515.8900 PI(18:1_20:4) 91220.5448 20610.0089 79366.3718 55342.3228 19467.9791 61214.7035 45953945.2000 38075525.8400 173939559.0000 PI(18:1_20:5) 18641.5958 21753.0150 36220.2603 218717.1455 24126.8755 150593.7052 4846131.9020 4289912.5730 14534862.5800 PI(18:1_22:5) 47521.9394 25238.3055 45461.8585 832186.6820 17941.0190 367496.4759 5735002.1200 3199873.6620 12060848.2800 PI(18:1e_18:1) 68011.1952 429083.4090 56770.2286 48032.7700 251687.0566 212406.7319 4239793.8760 1151718.9500 4424761.7290 PI(18:1e_20:5) 15875.6390 28810.9015 10630.1835 10373.3439 10524.7566 16968.7039 1170515.6830 943237.5834 2894220.2700 PI(18:1e_22:5) 19092.1062 30254.5392 24032.5402 38154.5445 15637.4401 72187.8601 2755810.0940 1951232.6730 7452694.8010 PI(19:0_20:3) 64505.2126 163692.0047 116713.0336 64223.3789 82038.6668 287771.5370 5715258.1400 3340603.6520 12081196.4800 PI(19:1_18:0) 33785.3798 50068.6905 63789.5743 133951.1616 57704.8386 244935.1091 2070446.1800 784889.5014 2763868.7120 PI(19:1_18:1) 71016.0510 114827.0677 114818.8133 332801.6748 55468.2059 322709.8034 12088468.2300 4249196.5640 16589332.0600 PI(32:2) 42207.9042 25399.6997 35625.2271 100028.7179 23737.2066 73058.6292 7176019.1880 4650969.3430 7646987.3440 PS(16:0_18:1) 132016.5500 452020.0300 37961.4600 100041.1400 529925.8100 PS(16:1_20:5) 23316.8946 26144.7013 24436.7962 24266.1249 24433.4454 24951.7057 2221939.0470 1461089.9090 2097249.3420 PS(18:0_20:3) 173600.1912 776214.5945 214976.6978 982480.4251 233887.6249 1035830.7820 9496314.3190 6509575.3240 12455556.3900 PS(18:1_18:2) 156121.7570 487985.5233 74011.0137 470452.2972 205321.7295 532845.5268 4482911.5410 2406105.1430 5372961.1950 PS(18:1_20:3) 163007.5693 401937.1315 199284.4415 1239707.7320 375088.0242 917943.4354 7592773.7250 6421396.4530 10462233.9800 PS(18:1_22:5) 37601.0360 103040.8572 73911.4817 2846977.0520 528013.6378 1921853.5430 3025386.2400 1495490.8230 3395737.0760 PS(18:2(9Z_12Z)/18:2(9Z_12Z)) 25717.9406 29072.0736 27537.0902 195738.4600 38158.8251 169543.7107 2641037.2190 990487.3576 1853561.8110 MS_METABOLITE_DATA_END #METABOLITES METABOLITES_START metabolite_name pubchem_id inchi_key kegg_id other_id other_id_type ri ri_type moverz_quant LPE(16:0) 7.813 LPE(18:0) 9.053 LPE(18:1) 8.064 LPE(18:2) 7.148 LPE(20:4) 7.065 LPE(20:5) 6.3 LPG(18:1) 6.861 LPG(20:0) 7.794 LPI(18:0) 7.547 PC(14:0_18:3) 12.705 PC(14:0_20:5) 12.504 PC(14:1e_18:2) 14.216 PC(14:1e_18:4) 12.643 PC(15:0_16:1) 13.963 PC(15:0_18:3) 13.052 PC(16:0_16:1) 14.496 PC(16:0_18:1) 12.56 PC(16:0_18:3) 13.672 PC(16:0e_20:5) 13.337 PC(16:1_14:0) 13.425 PC(16:1_16:1) 13.485 PC(16:1_18:1) 14.481 PC(16:1e_18:3) 14.381 PC(17:0_18:1) 15.224 PC(17:1_16:0) 15.07 PC(17:1_16:1) 14.064 PC(17:1_18:2) 14.264 PC(17:1_18:3) 13.66 PC(18:0_18:1) 13.46 PC(18:1_18:2) 14.878 PC(18:1_20:3) 14.97 PC(18:1e_22:5) 13.982 PC(18:1e_22:6) 12.938 PC(32:4e) 13.396 PC(34:6e) 12.992 PC(36:6e) 13.683 PC(36:7e) 13.15 PE(16:0_16:1) 12.983 PE(16:0_18:3) 12.347 PE(16:1_16:1) 12.172 PE(16:1_18:1) 13.014 PE(16:1e_16:1) 13.56 PE(16:1e_18:1) 14.656 PE(16:1e_18:2) 13.653 PE(16:1e_18:3) 12.89 PE(16:1e_20:3) 13.845 PE(16:1e_20:4) 13.225 PE(16:1e_20:5) 12.55 PE(16:1e_22:5) 13.489 PE(18:0_14:0) 13.04 PE(18:0_22:4) 13.634 PE(18:1_20:4) 12.941 PE(18:1_20:5) 12.155 PE(18:1_22:4) 13.645 PE(18:1_22:5) 12.067 PE(18:1_22:6) 12.398 PE(18:1e_18:2) 14.788 PE(18:1e_20:3) 14.955 PE(18:1e_20:4) 14.313 PE(18:1e_22:4) 15.17 PE(18:1e_22:5) 14.288 PE(18:2_18:2) 12.444 PE(18:2e_22:5) 13.819 PE(18:2e_22:6) 12.887 PE(18:3_18:2) 12.113 PE(20:5_18:2) 11.654 PE(33:2e) 14.098 PE(34:6e) 11.77 PE(38:7e) 12.792 PG(15:0_18:1) 11.107 PG(16:0_16:0) 11.346 PG(16:0_18:1) 11.352 PG(16:1_18:1) 10.955 PG(17:0_18:1) 11.587 PG(17:1_18:1) 11.16 PG(18:1_14:0) 10.862 PG(18:1_18:1) 11.393 PG(18:1_18:2) 11.024 PG(18:1_18:3) 10.252 PG(20:0_16:0) 11.467 PG(20:0_18:1) 11.504 PG(20:0_18:2) 11.162 PG(20:0_20:3) 11.155 PG(20:0_22:5) 11.033 PG(20:0_22:6) 10.888 PI(15:0_18:1) 11.02 PI(16:0_16:0) 11.229 PI(16:0_16:1) 10.764 PI(16:0_18:1) 11.267 PI(16:0_18:3) 10.459 PI(16:0_20:5) 10.363 PI(16:0e_20:3) 11.284 PI(16:0e_20:5) 10.682 PI(16:0e_22:5) 11.051 PI(16:1_18:1) 10.835 PI(17:0_18:1) 11.505 PI(17:0_20:3) 11.381 PI(17:0_20:4) 10.995 PI(17:0_20:5) 10.64 PI(18:0_18:1) 11.751 PI(18:0_20:1) 12.244 PI(18:0_20:2) 11.8 PI(18:0_20:3) 11.67 PI(18:0_20:4) 11.247 PI(18:0_22:2) 12.26 PI(18:0_22:3) 11.909 PI(18:0_22:4) 11.56 PI(18:0_22:5) 11.228 PI(18:0e_20:4) 11.549 PI(18:0e_22:5) 11.526 PI(18:1_18:1) 11.324 PI(18:1_18:2) 10.96 PI(18:1_18:3) 10.737 PI(18:1_20:4) 10.904 PI(18:1_20:5) 10.432 PI(18:1_22:5) 11.071 PI(18:1e_18:1) 11.663 PI(18:1e_20:5) 10.861 PI(18:1e_22:5) 11.366 PI(19:0_20:3) 11.707 PI(19:1_18:0) 11.963 PI(19:1_18:1) 11.604 PI(32:2) 12.55 PS(16:0_18:1) 2036611.74 PS(16:1_20:5) 12.545 PS(18:0_20:3) 15.094 PS(18:1_18:2) 13.979 PS(18:1_20:3) 14.088 PS(18:1_22:5) 13.663 PS(18:2(9Z,12Z)/18:2(9Z,12Z)) 13.017 METABOLITES_END #END