#METABOLOMICS WORKBENCH amaynard_20231001_190630 DATATRACK_ID:4356 STUDY_ID:ST002896 ANALYSIS_ID:AN004755 PROJECT_ID:PR001773 VERSION 1 CREATED_ON October 1, 2023, 7:11 pm #PROJECT PR:PROJECT_TITLE Folate depletion induces erythroid differentiation through perturbation of de PR:PROJECT_TITLE novo purine synthesis PR:PROJECT_SUMMARY Folate, an essential vitamin, is a one-carbon acceptor and donor in key PR:PROJECT_SUMMARY metabolic reactions. Erythroid cells harbor a unique sensitivity to folate PR:PROJECT_SUMMARY deprivation, as revealed by the primary pathological manifestation of PR:PROJECT_SUMMARY nutritional folate deprivation: megaloblastic anemia. To study this metabolic PR:PROJECT_SUMMARY sensitivity, we applied mild folate depletion to human and mouse erythroid cell PR:PROJECT_SUMMARY lines, and primary murine erythroid progenitors. We show that folate depletion PR:PROJECT_SUMMARY induces early blockade of purine synthesis and accumulation of the purine PR:PROJECT_SUMMARY synthesis intermediate and signaling molecule, AICAR, followed by enhanced heme PR:PROJECT_SUMMARY metabolism, hemoglobin synthesis, and erythroid differentiation. This is PR:PROJECT_SUMMARY phenocopied by inhibition of folate metabolism using the SHMT1/2 inhibitor - PR:PROJECT_SUMMARY SHIN1, and by AICAR supplementation. Mechanistically, the metabolically-driven PR:PROJECT_SUMMARY differentiation is independent of nucleotide sensing through mTORC1 and AMPK, PR:PROJECT_SUMMARY and is instead mediated by protein kinase C (PKC). Our findings suggest that PR:PROJECT_SUMMARY folate deprivation-induced premature differentiation of erythroid progenitor PR:PROJECT_SUMMARY cells is a molecular etiology to folate-deficiency induced anemia. PR:INSTITUTE Boston Children's Hospital, Harvard Medical School PR:DEPARTMENT pathology PR:LABORATORY Kanarek Lab PR:LAST_NAME Kanarek PR:FIRST_NAME Naama PR:ADDRESS Enders 1116.2, 300 Longwood Ave, Boston, MA 02115 PR:EMAIL naama.kanarek@childrens.harvard.edu PR:PHONE (617) 355-7433 #STUDY ST:STUDY_TITLE Polar metabolite levels in MEL cells following folate depletion and inosine ST:STUDY_TITLE supplementation ST:STUDY_SUMMARY Culture of MEL cells in RPMI media containing 100 nM folic acid for 48hr in the ST:STUDY_SUMMARY presence or absence of 100 uM inosine followed my metabolomics targeting polar ST:STUDY_SUMMARY metabolites. ST:INSTITUTE Boston Children's Hospital, Harvard Medical School ST:DEPARTMENT pathology ST:LABORATORY Kanarek Lab ST:LAST_NAME Kanarek ST:FIRST_NAME Naama ST:ADDRESS Enders 1116.2, 300 Longwood Ave, Boston, MA 02115 ST:EMAIL naama.kanarek@childrens.harvard.edu ST:PHONE 6173557433 ST:NUM_GROUPS 4 #SUBJECT SU:SUBJECT_TYPE Cultured cells SU:SUBJECT_SPECIES Mus musculus SU:TAXONOMY_ID 10090 SU:CELL_STRAIN_DETAILS MEL #SUBJECT_SAMPLE_FACTORS: SUBJECT(optional)[tab]SAMPLE[tab]FACTORS(NAME:VALUE pairs separated by |)[tab]Raw file names and additional sample data SUBJECT_SAMPLE_FACTORS MEL MEL_2000nM_FA_Vehicle_HILIC_1 Treatment:2000nM_FA_Vehicle RAW_FILE_NAME=Copy of 20220701_AM_KM_Inosine_Rescue_D2_AM805.raw SUBJECT_SAMPLE_FACTORS MEL MEL_2000nM_FA_Vehicle_HILIC_2 Treatment:2000nM_FA_Vehicle RAW_FILE_NAME=Copy of 20220701_AM_KM_Inosine_Rescue_D2_AM806.raw SUBJECT_SAMPLE_FACTORS MEL MEL_2000nM_FA_Vehicle_HILIC_3 Treatment:2000nM_FA_Vehicle RAW_FILE_NAME=Copy of 20220701_AM_KM_Inosine_Rescue_D2_AM807.raw SUBJECT_SAMPLE_FACTORS MEL MEL_100nM_FA_Vehicle_HILIC_1 Treatment:100nM_FA_Vehicle RAW_FILE_NAME=Copy of 20220701_AM_KM_Inosine_Rescue_D2_AM808.raw SUBJECT_SAMPLE_FACTORS MEL MEL_100nM_FA_Vehicle_HILIC_2 Treatment:100nM_FA_Vehicle RAW_FILE_NAME=Copy of 20220701_AM_KM_Inosine_Rescue_D2_AM809.raw SUBJECT_SAMPLE_FACTORS MEL MEL_100nM_FA_Vehicle_HILIC_3 Treatment:100nM_FA_Vehicle RAW_FILE_NAME=Copy of 20220701_AM_KM_Inosine_Rescue_D2_AM810.raw SUBJECT_SAMPLE_FACTORS MEL MEL_2000nM_FA_Inosine_HILIC_1 Treatment:2000nM_FA_Inosine RAW_FILE_NAME=Copy of 20220701_AM_KM_Inosine_Rescue_D2_AM811.raw SUBJECT_SAMPLE_FACTORS MEL MEL_2000nM_FA_Inosine_HILIC_2 Treatment:2000nM_FA_Inosine RAW_FILE_NAME=Copy of 20220701_AM_KM_Inosine_Rescue_D2_AM812.raw SUBJECT_SAMPLE_FACTORS MEL MEL_2000nM_FA_Inosine_HILIC_3 Treatment:2000nM_FA_Inosine RAW_FILE_NAME=Copy of 20220701_AM_KM_Inosine_Rescue_D2_AM813.raw SUBJECT_SAMPLE_FACTORS MEL MEL_100nM_FA_Inosine_HILIC_1 Treatment:100nM_FA_Inosine RAW_FILE_NAME=Copy of 20220701_AM_KM_Inosine_Rescue_D2_AM814.raw SUBJECT_SAMPLE_FACTORS MEL MEL_100nM_FA_Inosine_HILIC_2 Treatment:100nM_FA_Inosine RAW_FILE_NAME=Copy of 20220701_AM_KM_Inosine_Rescue_D2_AM815.raw SUBJECT_SAMPLE_FACTORS MEL MEL_100nM_FA_Inosine_HILIC_3 Treatment:100nM_FA_Inosine RAW_FILE_NAME=Copy of 20220701_AM_KM_Inosine_Rescue_D2_AM816.raw #COLLECTION CO:COLLECTION_SUMMARY One million cells from culture were collected via centrifugation for 20 seconds CO:COLLECTION_SUMMARY at 18,000xG, washed with 0.9% NaCl, and collected via centrifugation for 20 CO:COLLECTION_SUMMARY seconds at 18,000xG CO:SAMPLE_TYPE Cultured cells #TREATMENT TR:TREATMENT_SUMMARY MEL cells were cultured for up to 8 days in 100 nM folic acid containing RPMI TR:TREATMENT_SUMMARY media. The treatments were setup as a reverse time-course experiment and TR:TREATMENT_SUMMARY harvested on the same day. Day 0 samples were cultured for 8 days in 2,000 nM TR:TREATMENT_SUMMARY folic acid. #SAMPLEPREP SP:SAMPLEPREP_SUMMARY Pellet was resuspended in extraction buffer (80% Methanol, 25 mM Ammonium SP:SAMPLEPREP_SUMMARY Acetate and 2.5 mM Na-Ascorbate prepared in LC-MS water, supplemented with SP:SAMPLEPREP_SUMMARY isotopically-labelled amino acid standards [Cambridge Isotope Laboratories, SP:SAMPLEPREP_SUMMARY MSK-A2-1.2], aminopterin, and reduced glutathione standard [Cambridge Isotope SP:SAMPLEPREP_SUMMARY Laboratories, CNLM-6245-10]). Samples were vortexed for 10 sec, then centrifuged SP:SAMPLEPREP_SUMMARY for 10 minutes at 18,000 g to pellet cell debris. The supernatant was divided SP:SAMPLEPREP_SUMMARY into two tubes and dried on ice using a liquid nitrogen dryer. One tube of dried SP:SAMPLEPREP_SUMMARY sample was used for polar metabolite detection. Dried samples were resuspended SP:SAMPLEPREP_SUMMARY in 25 μL water #CHROMATOGRAPHY CH:CHROMATOGRAPHY_SUMMARY Sample was injected into a ZIC-pHILIC 150 x 2.1 mm (5 μm particle size) column CH:CHROMATOGRAPHY_SUMMARY (EMD Millipore). operated on a Vanquish™ Flex UHPLC Systems (Thermo Fisher CH:CHROMATOGRAPHY_SUMMARY Scientific, San Jose, CA, USA). Chromatographic separation was achieved using CH:CHROMATOGRAPHY_SUMMARY the following conditions: buffer A was acetonitrile; buffer B was 20 mM ammonium CH:CHROMATOGRAPHY_SUMMARY carbonate, 0.1% ammonium hydroxide in water; resulting pH is around 9 without pH CH:CHROMATOGRAPHY_SUMMARY adjustment. Gradient conditions we used were: linear gradient from 20% to 80% B; CH:CHROMATOGRAPHY_SUMMARY 20–20.5 min: from 80% to 20% B; 20.5–28 min: hold at 20% B at 150 mL/min CH:CHROMATOGRAPHY_SUMMARY flow rate. The column oven and autosampler tray were held at 25 °C and 4 °C, CH:CHROMATOGRAPHY_SUMMARY respectively. CH:CHROMATOGRAPHY_TYPE HILIC CH:INSTRUMENT_NAME Thermo Vanquish CH:COLUMN_NAME EMD Millipore ZIC-HILIC (100 x 2.1 mm,3.5 um) CH:SOLVENT_A 100% acetonitrile CH:SOLVENT_B 100% water; 20 mM ammonium carbonate; 0.1% ammonium hydroxide CH:FLOW_GRADIENT linear gradient from 20% to 80% B; 20–20.5 min: from 80% to 20% B; 20.5–28 CH:FLOW_GRADIENT min: hold at 20% B at 150 mL/min flow rate CH:FLOW_RATE 150 mL/min CH:COLUMN_TEMPERATURE 25 #ANALYSIS AN:ANALYSIS_TYPE MS #MS MS:INSTRUMENT_NAME Thermo Q Exactive Orbitrap MS:INSTRUMENT_TYPE Orbitrap MS:MS_TYPE ESI MS:ION_MODE UNSPECIFIED MS:MS_COMMENTS Polar metabolites were relatively quantified while referencing an in-house MS:MS_COMMENTS library of chemical standards and using TraceFinder 4.1 (Thermo Fisher MS:MS_COMMENTS Scientific, Waltham, MA, USA), with a 5 ppm mass tolerance. #MS_METABOLITE_DATA MS_METABOLITE_DATA:UNITS peak area MS_METABOLITE_DATA_START Samples MEL_2000nM_FA_Vehicle_HILIC_1 MEL_2000nM_FA_Vehicle_HILIC_2 MEL_2000nM_FA_Vehicle_HILIC_3 MEL_100nM_FA_Vehicle_HILIC_1 MEL_100nM_FA_Vehicle_HILIC_2 MEL_100nM_FA_Vehicle_HILIC_3 MEL_2000nM_FA_Inosine_HILIC_1 MEL_2000nM_FA_Inosine_HILIC_2 MEL_2000nM_FA_Inosine_HILIC_3 MEL_100nM_FA_Inosine_HILIC_1 MEL_100nM_FA_Inosine_HILIC_2 MEL_100nM_FA_Inosine_HILIC_3 Factors Treatment:2000nM_FA_Vehicle Treatment:2000nM_FA_Vehicle Treatment:2000nM_FA_Vehicle Treatment:100nM_FA_Vehicle Treatment:100nM_FA_Vehicle Treatment:100nM_FA_Vehicle Treatment:2000nM_FA_Inosine Treatment:2000nM_FA_Inosine Treatment:2000nM_FA_Inosine Treatment:100nM_FA_Inosine Treatment:100nM_FA_Inosine Treatment:100nM_FA_Inosine 2-hydroxybutyric acid 1688124.2 1920787.272 1753371.704 1891652.379 2237355.543 1887096.941 2033273.789 2086949.464 2233483.852 2469181.695 2975002.665 1984512.185 2-hydroxyglutarate 1374463967 1789634243 1579931147 1263715734 1849843600 2130426506 2110726415 1961406253 1841907237 2638896016 1901906834 2606944609 2-oxobutyric acid 58492926.21 55931666.29 52458175.33 50577133.74 57683781.8 52944612.15 57418181.37 59278620.29 60991616.43 58241094.86 74714153.79 66401694.93 2,3-BPG 3978946.201 3151312.4 3106265.267 2937209.335 3337495.086 4279468.835 2177140.788 412708.6467 1704766.398 1475408.807 2903766.27 1710973.149 3-methyl-2-oxopentanoic acid 65832197.74 71492268.48 55773318.49 51386341.38 52707494.22 51783347.26 55706568.57 44822449.17 52512781.4 51490744.87 49082170.71 46291219.82 3-phosphoglycerate 23984311.61 25930548.59 23224866.94 23888944.39 25423310.36 25936948.13 20526545.85 20835860.15 19895136.65 23834723.14 23870739.4 22202306.64 6-phosphogluconate 7078894.062 6751841.393 7085747.263 5834111.457 6437227.92 6609875.739 5788626.419 4936461.179 5864905.891 4772867.684 5296643.21 5260698.538 Acetyl-CoA 31747661.66 28119701 29504566.94 29068762.13 26808604.39 26678407.92 30849813.49 29279016.97 33647177.79 28342022.96 37999014.32 32664408.4 acetylcarnitine 54442767.04 54050540.15 57606908.85 53282464.75 55004237.12 58019414.59 123167778.2 122326771.3 133958407.7 146360986.7 139644439.5 140475524.8 ADP 44470799.01 34989925.45 56028005.44 34877732.89 21971897.32 22520725.18 33475954.31 48907252.2 36344907.78 30371886.94 48236988.64 29618618.35 alanine 467700467.6 429179762.8 434344623.3 328574018.3 312331053.2 343525180.7 256919799.9 236959337.8 232294273.2 230466629.8 225547497.6 215459206.6 Alpha-Ketoglutarate 46927311.97 44028986.21 43244157.32 36955147.87 45980083.15 40209722.8 39978017.21 50853835.84 30925553.15 23737168.69 28942427.7 38044799.78 Alpha-Lipoic Acid 52562957.23 53134551.25 39139733.1 52556005.11 52214417.34 43520243.63 46023189.42 42352861.71 46593733.76 40877850.65 66364673.63 48111132.09 AMP 7250141.613 8670088.353 7217598.398 6567310.34 6829675.387 7476382.315 6179715.857 8582117.41 6657957.287 6287862.891 5500203.839 5631801.668 arginine 3671569867 3828566471 4382358283 3451684300 3465690516 3080067736 3421944652 4165738144 3264061115 3252725219 3382471298 2939203182 argininosuccinate 54392788.48 56017001.78 61406284.91 33695427.33 35887328.72 36681014.72 32203684.92 29317527.67 26186039.76 27195589.52 27982536.65 28536716.25 asparagine 1722785913 1680391768 1550158810 1435568881 1474468090 1577671727 1501270066 1547852463 1520165741 1461473386 1540523158 1436410467 aspartate 187228730.6 183199249 159306939.7 162557881.6 170702009.7 187336215.2 154402410.4 136312668.9 161686385.2 166334456.5 147281763.9 151207164.4 ATP 566255020.7 424795451.2 564787510.1 475513461.3 311726405.5 339054137.7 416680718.2 603943129.4 458744241.4 397113013.9 641550652 408046921.7 beta-alanine 122201274.8 107938720.4 125232928.5 99019070.87 107502689.8 115786814.4 126059632.3 129374740 136685595.6 115942302.4 136263924.7 114420084.5 Butyrylcarnitine 48020972.97 51518397.46 48412314.02 50322232.61 52066084.62 52820711.36 56506243.88 47937905.25 58708029.16 59447545.99 61519443.15 61038069.06 C5-Carnitines 3919106.57 3633672.703 4867464.135 1973656.901 2465686.127 2981738.253 1404556.869 2205463.209 1214396.033 1883945.272 1287903.474 1459417.229 carnitine 180511926.7 172598104.2 176483659.2 148044359.8 154161608.7 157451590.7 175632684.1 163117546.2 171617000.9 163554292.8 168308097.9 158595473.9 CDP-choline 2067542.45 1997857.668 2426583.082 6052978.971 6092034.692 5446421.556 2682804.851 2980966.489 2741305.034 3072247.739 3600970.585 3197178.306 cis-aconitate 48716281 44572884.47 49555530.73 52675010.87 51065543.03 47521106.24 43743577.27 43671121.25 47834967.41 37606114.54 42046782.72 40720552.22 citrate 1365483913 1276418425 1393090378 1432835947 1420288733 1344036310 1085132061 1174357728 1151343218 1039533692 1221449744 1098714141 Coenzyme-A 13100689.83 13261836.04 13734486.69 12401386.75 14585720.74 12876973.67 16188653.71 15686193.57 15906415.59 17199894.69 14795670.83 17106892.18 creatine 79279339.61 81847232.59 106715721.1 85754985.97 97562984.49 98606954.63 88552365.69 100194070.4 81828976.65 83645458.6 94686945.33 83062712.89 CTP 64463843.8 52262038.22 59207632.28 60418411.48 39728788.33 42423218.43 18352015.08 26004038.36 19524854.73 18879674.06 31513921.38 19721145.96 dATP 3210306.224 2516348.525 4335431.164 2402948.047 1511734.525 1517817.384 3231445.489 4511394.677 3763204.616 3550358.455 5621672.143 3632771.322 dCTP 10305546.36 8167910.833 8458840.463 31205911.52 19248178.75 19706203.21 2997083.776 4262316.875 3187225.629 3374567.347 5718316.575 3666574.738 dihydroxyacetonephosphate 10800153.72 13950927.38 12441167.19 9369118.727 12943581.26 15338727.11 26365352.36 24213337.96 25257613.57 34068784.33 18956979.87 34726572.89 erythrose-4-phosphate 1557200.68 1380367.257 1737245.048 1041463.233 938453.9608 1049370.057 1009464.754 841508.2304 1186277.427 799365.0477 650431.7317 700221.2524 F6P G1P 33205170.34 33515289.94 36159326.36 27161123.2 28364318.68 26728304.25 57595211.4 60933200.53 64001586.28 54107437.33 56563293.34 56152178.36 folic acid 773959.2536 782280.6467 681283.018 26745.47595 971583.774 870210.4078 901749.9109 17767.14513 formyl methionine 25801262.2 26156054.22 18207574.82 20337015.23 20081682.15 19554403.85 24002798.53 16853681.09 25820815.7 22627494.34 23533752.06 23182029.47 Fructose-1,6-bisphosphate 36056645.31 36830420.46 39739533.04 35505457.07 33676898.75 36663325.26 55781353.4 64463105.11 66248714.92 65638673.03 69381884.95 75875030 fumarate 102920864.2 103252649.9 96456373.87 80841157.82 85261825.59 86478237.82 81033928.85 65452902.77 69277872.4 70483603.28 66919793.89 69325971.21 GAR 25539401.26 26492010.05 20973260.67 21451937.63 20288060.86 20204806.5 GDP 3640130.01 3483753.66 3536664.414 3495224.917 3625731.092 3672990.344 4218907.959 4578265.199 4222739.861 4305843.679 3955477.599 4296163.363 Glucose-6-phosphate 41380600.01 40018428.52 41228603.01 31260114.15 33730925.31 30452543.42 30611915.48 26424051.66 30998689.13 23852670.3 25214124.85 24367858.42 glutamate 3467290583 3654443946 3477024298 3825188756 4189460418 4346106679 3983037521 3425140264 4155144074 4016558556 3777299203 3905945590 glutamine 6250658363 6116287194 7276041718 5777065541 6334161978 6686880471 7147904664 7878145406 6834574681 7094465498 8001557830 7169243782 glycerate 246849000.9 261235282.9 284259954.2 227193229 274377085.1 263561457.5 276352851.8 305871633.4 280587965.3 297587353.8 341946828.9 332287719.5 Glycerol-3-Phosphate 2503168.09 1280801.527 1057931.534 2436826.88 720993.9884 4174323.386 3646210.39 4208950.266 2055977.141 2761152.217 3995341.877 Glycerophosphocholine 20590586889 20527815064 23138892674 14631261959 13998709257 15232842544 23094679183 25768068199 22738956955 22890124818 24204397322 21468290846 glycine 66271456.61 56483027.38 55492496.99 39979536.53 45034460.42 49505272.47 90545780.43 87710595.17 95580582.36 86983310.92 78911070.41 87816872.52 GSH 4731698656 4916107834 3310254678 5071378513 5441392326 5675961830 5254459885 3638180218 5016955307 5542764206 5864152552 5645329475 hydroxyproline gammaALA 277840541.8 274374093.2 240422560.6 198488561 201579025.5 233998842.2 253002868.5 202859561 264820886.1 234246437.2 216509037.1 227473353.9 hypotaurine 2157557.086 2203074.982 2851930.364 2590183.756 2690100.2 2826531.17 2244247.051 2502758.696 2148829.655 2626397 2747268.183 3351053.837 hypoxanthine 2979574.522 6310805.504 17355907.24 3457547.537 3638799.771 3170153.34 434585395.3 387906708.1 334406802.5 739864411.6 618546529.3 535087839.1 IMP 10549382.13 11642421.94 12548970.62 16623091.6 19590367.64 20933727.34 10099686.86 11584488.19 11506954.51 11351260.32 12833823.47 12204410.95 leucine 1055558119 1036499710 1162920114 861667892.2 952128606.9 953420942.7 1168638712 1088120991 1129901404 1187078441 1021682274 1013808327 lysine 26804077.46 28656386.39 35714999.87 27960779.13 35844516.08 30050913.19 13930574.21 18169707.55 13662451.53 18554216.56 17579011.8 13963992.75 malate 1141469780 1160958937 1080861039 887603306.1 930338909.7 958371014 799846954 664365979 741954619.7 717676602.3 745762867.8 703387647.9 malonyl-CoA 1867330.31 1362060.554 1780663.119 1665829.78 2086147.124 2049009.196 4440360.251 4452420.687 4640319.281 4455882.638 4926549.583 5169078.121 methionine 331216555.1 363236212.4 361329062.6 291334169.4 333965362.3 410991210.2 373380162.4 346460472.7 330086827 588692510.4 315873331 430258831.7 methylthioadenosine 50784937.55 57316211.49 82075467.51 63133544.18 74302411.31 74619079.25 71498594.68 88535021.17 67357463.29 81292559.32 73455592.45 80317354.58 N-acetylaspartate 967022.095 308938.2689 198704.7359 2551710.731 315591.9956 N-acetylmethionine 1570733.707 2525904.075 1486068.783 1609453.92 1698707.271 2122461.197 2487615.06 2679133.277 3086965.32 3479657.062 2758146.417 3761307.911 N-acetylphenylalanine 947813.3193 1077135.32 674225.4667 711146.4638 878187.1765 919084.1106 1488896.614 818606.8366 1560655.993 1593912.981 1478416.458 NAD. 546222388.6 549755214.2 558171216.6 484766744.6 497490114.2 547222471.2 535330011.9 548162877.9 537866014.7 467309483.6 555385650.7 523831270.1 NADP 16034498.91 17718274.16 18536269.73 14202988.29 16405189.62 15452109.58 14348074.4 17391903.49 14373383.5 13643881.64 12703504.52 15157936.24 niacinamide 414714797.2 463132747.1 553930603.3 432313644.3 535143624.3 462933918.2 1943923393 1790154494 1538937362 2618337343 1758362918 1896099279 O-phosphoethanolamine 439741.4517 572674.9368 558111.7814 780097.7464 685171.8477 541051.7215 677483.7984 488523.0916 562603.4297 414539.4925 388208.0851 508049.712 Pantothenate 631201212.8 650010335.8 676733826.3 612629268.8 621505384.8 621613501.2 582777257.6 594026706 597574484.5 568373401.7 657461335.4 587066384.2 phenylalanine 48669690.41 52186675.97 53857801.09 50366700.88 57828238.95 59206615.8 37375704.17 33662205.43 39412803.07 50786737.71 47750354.95 43690484.69 phosphocholine 16302085062 15686078698 18646649389 15001939787 15580625150 15087153761 16513624444 20328825586 16135868761 17803508137 20275245559 18455552516 phosphocreatine phosphoenolpyruvate 9852157.695 10287952.41 8718484.42 10423767.11 10411977.53 9693201.332 6863573.208 6731656.751 6276498.286 7147967.663 6853091.007 5905871.895 proline 13515186603 12820397940 15933554832 11720607296 11587833845 11036666302 14753970725 15731491644 13789096826 13480350355 15321457835 12970194627 propionylcarnitine 30803920.47 27937232.19 21682893.2 14117452.72 15736224.11 17898144.47 10168076.5 6011635.192 9936292.602 9327054.098 8424979.907 9061744.112 PRPP 11752101.72 14192014.59 9211477.407 22967203.64 18376296.11 19117099.56 1332131.016 1263748.165 1553784 1273619.021 1304199.578 Pyridoxine 283623437.8 321764195.2 293187464.2 307790921.9 315120615.7 304154091.8 351662965.4 332420578.9 341988605.4 372020530.6 347410809.2 308504707.2 pyruvate 16215070.34 17614743.06 13222182 11664025.88 7710926.54 7694753.363 14639400.96 22347102.62 10438031.51 10735645.64 6896446.75 12115546.02 quinolinic acid 786737.6322 1136938.426 767023.905 985061.545 955517.5043 892492.6267 985322.0812 904125.3603 1038808.068 878454.9367 1438501.484 1260855.686 Ribose-1-phosphate 2091429.052 2415436.793 2079344.127 4900232.519 5319910.227 4643673.566 633560.3874 675386.0852 721803.8585 639578.5153 534285.7118 615526.2192 Ribose.Ribulose.5.phosphate.3Me.Lys 2795305.318 3389748.202 3276857.146 3400717.844 3605852.666 3948715.361 2682857.873 2181996.05 2465045.638 2907673.477 1826811.532 2710338.499 S-Adenosyl-L-Homocysteine 10849485.87 10975179.64 12508499.96 11563018.48 11993936.77 12450325.01 9370120.642 11137080.26 9561573.864 9522302.571 10123530.88 10824486.86 Sedoheptulose-7-phosphate 30533587.61 31411756.12 36190935.23 21899526.93 21989278.96 19989181.53 77113989.22 76188157.38 85082955.66 73921449.05 75617373.29 75668988.83 serine 9994660.1 12037616.83 10005492.51 27682208.57 41478632.44 49730616.81 14457292.64 17433386.16 16527296.29 65667776.17 59497649.73 59318337.03 succinate 62426692.82 66450937.78 74722582.59 48713852.39 62728810.05 67147609.59 75155894.14 69206721.66 70545157.38 82102975.4 59399507.71 79898713.56 taurine 8775889.35 8828150.712 10093057.19 8530603.794 11612172.29 11471157.08 8767489.841 9192779.011 8754312.031 9434326.732 8958426.543 10613618.88 threonine 263260303.6 229820982.2 214990857 323830676.5 259081146.2 213189094.6 185581033.2 165724182.2 220804383.7 182955490.3 292159616.5 162779111.8 tryptophan 74983291.22 80656485.04 79981547.69 68669144.09 84203238.01 103547821.2 61863770.31 55926737.3 57568452.47 94497199.85 67128741.59 78334513.5 tyrosine 408989386.5 419856618.6 434851895.7 323738696.8 362288994.4 361887430.6 394188751.5 409431549.1 368013185 384639885.3 375834730 371267391.6 UDP.GlcNAc 220884614.6 225797905.6 197445341.5 156234115.5 178465544.3 174272719.5 143680853 138243057.1 155988395.9 139835448 147987752 154679981.4 UDP 7399873.746 7149344.783 9405302.389 7251896.493 7610971.902 7192808.291 3018520.124 4653391.055 3075195.374 3137480.29 3776071.705 3445312.395 UMP 1070747.498 1200188.794 1209399.473 1166685.219 1370176.463 1300518.817 811048.1488 724694.6814 837609.0495 965707.6838 1053970.458 973030.0331 Ureidosuccinic acid 1069645.314 1436090.693 852600.5038 2013311.96 2079940.817 2050681.705 1429145.366 1700731.192 1619043.091 1723882.727 2071839.455 2253458.048 UTP 353945768.6 287548784.1 315461583 376822598.5 270517108.3 278451417.1 93860640.47 131136942.3 107561956.1 97023796.52 154346500.4 104147076.1 xanthine 448256.0679 427408.1478 420705.1598 386782.1044 439222.2285 450897.3364 3968611.281 3557700.565 3394638.129 3414518.965 3026990.258 3228371.869 xanthosine 130344.625 120824.0239 135706.0422 97010.5343 171756.9439 170005.5972 276145.8174 279136.8481 287073.9453 299378.9637 302627.7576 291295.6787 MS_METABOLITE_DATA_END #METABOLITES METABOLITES_START metabolite_name Standardized name Formula Exact mass Sub class 2-hydroxybutyric acid 2-Hydroxybutyric acid C4H8O3 104.0473 Hydroxy FA 2-hydroxyglutarate 2-Hydroxyglutaric acid C5H8O5 148.0372 Dicarboxylic acids 2-oxobutyric acid 3-Methyl pyruvic acid C4H6O3 102.0317 Oxo FA 2,3-BPG - - - - 3-methyl-2-oxopentanoic acid 3-Methyl-2-oxovaleric acid C6H10O3 130.0630 Short-chain keto acids 3-phosphoglycerate 3-Phosphoglyceric acid C3H7O7P 185.9929 Short-chain acids 6-phosphogluconate 6-Phosphogluconic acid C6H13O10P 276.0246 Monosaccharide phosphates Acetyl-CoA Acetyl-CoA C23H38N7O17P3S 809.1258 Acyl CoAs acetylcarnitine CAR 2:0 C9H17NO4 203.1158 Acyl carnitines ADP ADP C10H15N5O10P2 427.0294 Purine rNDP alanine Alanine C3H7NO2 89.0477 Amino acids Alpha-Ketoglutarate Oxoglutaric acid C5H6O5 146.0215 TCA acids Alpha-Lipoic Acid Lipoic acid C8H14O2S2 206.0435 Thia FA AMP AMP C10H14N5O7P 347.0631 Purine rNMP arginine Arginine C6H14N4O2 174.1117 Amino acids argininosuccinate Argininosuccinic acid C10H18N4O6 290.1226 Amino acids asparagine Asparagine C4H8N2O3 132.0535 Amino acids aspartate Aspartic acid C4H7NO4 133.0375 Amino acids ATP ATP C10H16N5O13P3 506.9958 Purine rNTP beta-alanine beta-Alanine C3H7NO2 89.0477 Amino acids Butyrylcarnitine CAR 4:0 C11H21NO4 231.1471 Acyl carnitines C5-Carnitines - - - - carnitine Carnitine C7H15NO3 161.1052 Carnitines CDP-choline CDP-choline C14H26N4O11P2 488.1073 Pyrimidine rNDP cis-aconitate cis-Aconitic acid C6H6O6 174.0164 TCA acids citrate Citric acid C6H8O7 192.0270 TCA acids Coenzyme-A Coenzyme A C21H36N7O16P3S 767.1152 Other purines creatine Creatine C4H9N3O2 131.0695 Amino acids CTP CTP C9H16N3O14P3 482.9845 Pyrimidine rNTP dATP dATP C10H16N5O12P3 491.0008 Purine dNTP dCTP dCTP C9H16N3O13P3 466.9896 Pyrimidine dNTP dihydroxyacetonephosphate Dihydroxyacetone phosphate C3H7O6P 169.9980 Short-chain acids erythrose-4-phosphate Erythrose 4-phosphate C4H9O7P 200.0086 Monosaccharide phosphates F6P G1P - - - - folic acid Folic acid C19H19N7O6 441.1397 Folic acids formyl methionine N-Formylmethionine C6H11NO3S 177.0460 Amino acids Fructose-1,6-bisphosphate Fructose 1,6-bisphosphate C6H14O12P2 339.9961 Monosaccharide phosphates fumarate Fumaric acid C4H4O4 116.0110 TCA acids GAR GAR C7H15N2O8P 286.0566 Glycinamide ribonucleotides GDP GDP C10H15N5O11P2 443.0243 Purine rNDP Glucose-6-phosphate Glucose 6-phosphate C6H13O9P 260.0297 Monosaccharide phosphates glutamate Glutamic acid C5H9NO4 147.0532 Amino acids glutamine Glutamine C5H10N2O3 146.0691 Amino acids glycerate Glyceric acid C3H6O4 106.0266 Sugar acids Glycerol-3-Phosphate sn-Glycero-3-phosphate C3H9O6P 172.0137 Organic phosphoric acids Glycerophosphocholine sn-Glycero-3-phosphocholine C8H20NO6P 257.1028 Organic phosphoric acids glycine Glycine C2H5NO2 75.0320 Amino acids GSH Glutathione C10H17N3O6S 307.0838 Tripeptides hydroxyproline gammaALA - - - - hypotaurine Hypotaurine C2H7NO2S 109.0198 Sulfinic acids hypoxanthine Hypoxanthine C5H4N4O 136.0385 Hypoxanthines IMP IMP C10H13N4O8P 348.0471 Purine rNMP leucine Leucine C6H13NO2 131.0946 Amino acids lysine Lysine C6H14N2O2 146.1055 Amino acids malate D-Malic acid C4H6O5 134.0215 Beta hydroxy acids malonyl-CoA Malonyl-CoA C24H38N7O19P3S 853.1156 Acyl CoAs methionine Methionine C5H11NO2S 149.0511 Amino acids methylthioadenosine Methylthioadenosine C11H15N5O3S 297.0896 Purine ribonucleosides N-acetylaspartate N-Acetylaspartic acid C6H9NO5 175.0481 Amino acids N-acetylmethionine N-Acetylmethionine C7H13NO3S 191.0616 Amino acids N-acetylphenylalanine N-Acetylphenylalanine C11H13NO3 207.0895 Amino acids NAD. NAD+ C21H27N7O14P2 663.1091 Nicotinamide dinucleotides NADP NADP+ C21H28N7O17P3 743.0755 Nicotinamide dinucleotides niacinamide Niacinamide C6H6N2O 122.0480 Pyridine alkaloids O-phosphoethanolamine Phosphoethanolamine C2H8NO4P 141.0191 Phosphoethanolamines Pantothenate Pantothenic acid C9H17NO5 219.1107 Amino acids phenylalanine Phenylalanine C9H11NO2 165.0790 Amino acids phosphocholine Choline phosphate C5H14NO4P 183.0660 Cholines phosphocreatine Phosphocreatine C4H10N3O5P 211.0358 Amino acids phosphoenolpyruvate Phosphoenolpyruvic acid C3H5O6P 167.9824 Short-chain acids proline Proline C5H9NO2 115.0633 Amino acids propionylcarnitine CAR 3:0 C10H19NO4 217.1314 Acyl carnitines PRPP Phosphoribosyl pyrophosphate C5H13O14P3 389.9518 Monosaccharide phosphates Pyridoxine Pyridoxine C8H11NO3 169.0739 Pyridine alkaloids pyruvate Pyruvic acid C3H4O3 88.0160 Short-chain acids quinolinic acid Quinolinic acid C7H5NO4 167.0219 Pyridine alkaloids Ribose-1-phosphate Ribose 1-phosphate C5H11O8P 230.0192 Monosaccharide phosphates Ribose.Ribulose.5.phosphate.3Me.Lys - - - - S-Adenosyl-L-Homocysteine S-Adenosylhomocysteine C14H20N6O5S 384.1216 Other purines Sedoheptulose-7-phosphate Sedoheptulose 7-phosphate C7H15O10P 290.0403 Monosaccharide phosphates serine Serine C3H7NO3 105.0426 Amino acids succinate Succinic acid C4H6O4 118.0266 TCA acids taurine Taurine C2H7NO3S 125.0147 Sulfonic acids threonine Threonine C4H9NO3 119.0582 Amino acids tryptophan Tryptophan C11H12N2O2 204.0899 Amino acids tyrosine Tyrosine C9H11NO3 181.0739 Amino acids UDP.GlcNAc UDP-N-acetylglucosamine C17H27N3O17P2 607.0816 Pyrimidine ribonucleosides UDP UDP C9H14N2O12P2 404.0022 Pyrimidine rNDP UMP UMP C9H13N2O9P 324.0359 Pyrimidine rNMP Ureidosuccinic acid N-Carbamoylaspartic acid C5H8N2O5 176.0433 Amino acids UTP UTP C9H15N2O15P3 483.9685 Pyrimidine rNTP xanthine Xanthine C5H4N4O2 152.0334 Xanthines xanthosine Xanthosine C10H12N4O6 284.0757 Purine ribonucleosides METABOLITES_END #END