#METABOLOMICS WORKBENCH amaynard_20231001_184733 DATATRACK_ID:4355 STUDY_ID:ST002895 ANALYSIS_ID:AN004754 PROJECT_ID:PR001773 VERSION 1 CREATED_ON October 1, 2023, 6:55 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 K562 cells following folate depletion and inosine ST:STUDY_TITLE supplementation ST:STUDY_SUMMARY Culture of K562 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 Homo sapiens SU:TAXONOMY_ID 9606 SU:CELL_STRAIN_DETAILS K-562 #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 K562 K562_2000nM_FA_Vehicle_HILIC_1 Treatment:2000nM_FA_Vehicle RAW_FILE_NAME=Copy of 20220701_AM_KM_Inosine_Rescue_D2_AM793.raw SUBJECT_SAMPLE_FACTORS K562 K562_2000nM_FA_Vehicle_HILIC_2 Treatment:2000nM_FA_Vehicle RAW_FILE_NAME=Copy of 20220701_AM_KM_Inosine_Rescue_D2_AM794.raw SUBJECT_SAMPLE_FACTORS K562 K562_2000nM_FA_Vehicle_HILIC_3 Treatment:2000nM_FA_Vehicle RAW_FILE_NAME=Copy of 20220701_AM_KM_Inosine_Rescue_D2_AM795.raw SUBJECT_SAMPLE_FACTORS K562 K562_100nM_FA_Vehicle_HILIC_1 Treatment:100nM_FA_Vehicle RAW_FILE_NAME=Copy of 20220701_AM_KM_Inosine_Rescue_D2_AM796.raw SUBJECT_SAMPLE_FACTORS K562 K562_100nM_FA_Vehicle_HILIC_2 Treatment:100nM_FA_Vehicle RAW_FILE_NAME=Copy of 20220701_AM_KM_Inosine_Rescue_D2_AM797.raw SUBJECT_SAMPLE_FACTORS K562 K562_100nM_FA_Vehicle_HILIC_3 Treatment:100nM_FA_Vehicle RAW_FILE_NAME=Copy of 20220701_AM_KM_Inosine_Rescue_D2_AM798.raw SUBJECT_SAMPLE_FACTORS K562 K562_2000nM_FA_Inosine_HILIC_1 Treatment:2000nM_FA_Inosine RAW_FILE_NAME=Copy of 20220701_AM_KM_Inosine_Rescue_D2_AM799.raw SUBJECT_SAMPLE_FACTORS K562 K562_2000nM_FA_Inosine_HILIC_2 Treatment:2000nM_FA_Inosine RAW_FILE_NAME=Copy of 20220701_AM_KM_Inosine_Rescue_D2_AM800.raw SUBJECT_SAMPLE_FACTORS K562 K562_2000nM_FA_Inosine_HILIC_3 Treatment:2000nM_FA_Inosine RAW_FILE_NAME=Copy of 20220701_AM_KM_Inosine_Rescue_D2_AM801.raw SUBJECT_SAMPLE_FACTORS K562 K562_100nM_FA_Inosine_HILIC_1 Treatment:100nM_FA_Inosine RAW_FILE_NAME=Copy of 20220701_AM_KM_Inosine_Rescue_D2_AM802.raw SUBJECT_SAMPLE_FACTORS K562 K562_100nM_FA_Inosine_HILIC_2 Treatment:100nM_FA_Inosine RAW_FILE_NAME=Copy of 20220701_AM_KM_Inosine_Rescue_D2_AM803.raw SUBJECT_SAMPLE_FACTORS K562 K562_100nM_FA_Inosine_HILIC_3 Treatment:100nM_FA_Inosine RAW_FILE_NAME=Copy of 20220701_AM_KM_Inosine_Rescue_D2_AM804.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 K562 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 K562_2000nM_FA_Vehicle_HILIC_1 K562_2000nM_FA_Vehicle_HILIC_2 K562_2000nM_FA_Vehicle_HILIC_3 K562_100nM_FA_Vehicle_HILIC_1 K562_100nM_FA_Vehicle_HILIC_2 K562_100nM_FA_Vehicle_HILIC_3 K562_2000nM_FA_Inosine_HILIC_1 K562_2000nM_FA_Inosine_HILIC_2 K562_2000nM_FA_Inosine_HILIC_3 K562_100nM_FA_Inosine_HILIC_1 K562_100nM_FA_Inosine_HILIC_2 K562_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 3208167.405 3417997.593 3453362.316 2777836.575 2505461.421 2379756.969 3317568.136 3585722.828 3540124.131 4045274.332 3166420.151 3358720.986 2-hydroxyglutarate 1176531769 1059873834 1099901554 1177419977 1093722514 1234811936 1063046060 1132674327 1082752409 1122436502 1268625699 1046134687 2-oxobutyric acid 39854009.95 49000744.98 45546164.77 44140219.38 44477736.16 50141083.49 46836787 44342353.57 40182513 43155047.34 51804401.77 47424067.55 2,3-BPG 24510761.41 25762527.4 25561335.47 17980351.38 18528060.57 18921455.39 28373622.16 24826798.41 26713251.81 27916530.36 28464914.91 27817838.95 3-methyl-2-oxopentanoic acid 126642172 140602104.4 139750270.9 83703620.3 93463734.09 72285921.2 124670063.3 115783220.7 124090140.2 140938726.6 122829937.8 112851817.2 3-phosphoglycerate 40481799.69 40922123.76 44457486.84 31638065.76 30913425.25 33633731.05 38951476.73 36897580.79 36295152.87 37322686.6 39430468.58 37894397.36 6-phosphogluconate 19329161.03 20187754.07 18568356.03 12998572.22 12612414.25 11682489.89 16326955.92 16719463.58 15253371.89 16380131.77 16441543.67 14902515.71 Acetyl-CoA 17814070.84 17860707.65 17388804.28 15032009.01 15401203.71 14579797.39 18603967.62 19007748.26 18036751.39 17702182.66 18145384.02 19138892.66 acetylcarnitine 179566368 166981770.2 183951727.9 238885487 213117761.6 232235743.5 208578914 194051179.7 205483539.2 231379580.5 204919633.5 210077873.7 ADP 24873233.14 32794991.8 26453942.47 21361902.23 25850644.93 18087774.45 53053924.24 55388160.6 54462214.64 37110423.35 38665106.35 51107398.82 alanine 33492901.81 32547271.51 38335042.63 39005866.42 45510999.23 46132689.61 26519863.91 33949776.88 33717157.38 29199770.87 31470702.44 30866979.16 Alpha-Ketoglutarate 37154471.37 41600390.07 39328567.08 31681656.63 26279448.63 31836929.86 30974833.85 34048040.78 30106853.16 34857028.65 38714052.08 37036681.07 Alpha-Lipoic Acid 33931671.3 39819813.14 37035521.29 32618113.67 32871257.32 35087154.54 29938369.21 25465721.04 25796370.68 28420636.53 35313893.35 32286098.69 AMP 4419559.559 4212682.678 4117679.041 3351117.757 3654088.457 3574853.29 3933498.397 3473416.006 3406089.913 4358240.511 4147985.841 4712635.153 arginine 2261218868 2473240857 2397513256 2541818326 2748658955 2533345885 2536977350 2786994297 2960924773 2462899601 2347970020 2617248707 argininosuccinate 40515049.8 36812935.83 38242710.65 33029313.11 28335611.42 31750969.69 31969232.25 32074544.87 31012602.76 31623916.97 31627569.15 35845861.55 asparagine 995870521.6 950727918 959653901.9 1012742258 1017981641 1039079851 839152978.5 850331988.1 816452370.2 939959160.9 912396131.7 848739317.7 aspartate 214577813.1 188893566.5 198310296.5 268354646.7 254539094.4 276804232.3 195182659.1 173932095 183080561.9 226170538.6 214320570.8 193869381.8 ATP 325327269.9 399660151.9 328251386 280678569.7 353639848.1 236272535 606009802 564413194.2 552836017.5 404944607.6 449402232.1 622537388 beta-alanine 185046657.7 165891147.7 179744013.3 172640328.2 169366404.4 181987810.8 159644970.1 165464556.4 152792840.4 158362089.4 162826671.9 162513174 Butyrylcarnitine 23366264.34 22801734.61 24356207.14 20622393.43 18411473.33 20528789.86 19221599.59 20750509 21399021.65 22179755.99 20284773.94 19661760.29 C5-Carnitines 149639915.6 139836819.8 151611609.5 169669274.9 146623561.4 171245154.5 118775440.4 114429904.8 124940493.1 130590406.8 124477569 126005227.5 carnitine 196954595.9 176081655.1 196194990.1 164684537.7 163058923.2 169712342.9 166962488.3 171842114.9 173807909.1 172461379.4 161585695.1 162990685.2 CDP-choline 964662.3112 650279.0066 866781.9569 582546.833 788684.5103 670797.5022 900941.5314 711616.9966 651109.259 797874.6299 843872.3426 820765.2981 cis-aconitate 26440510.86 24618821.4 26186648.82 28424504.68 27390270.41 27098014.02 24031999.7 27397948.27 26147933.64 25445734.62 24909780.31 26284315.84 citrate 764307094.3 764615030.5 767780478.9 910300235.7 811823753.1 837739638.8 727766138.3 800987628.2 777085794.9 760826455.2 748854503.1 748684152.6 Coenzyme-A 5529453.066 6100720.704 8293395.945 6481675.366 7021450.895 7174045.867 4925484.318 8549628.61 8417812.409 7432296.559 8260704.623 7507671.555 creatine 1071779849 1163464420 1230012950 1396363666 1383195589 1462709711 948809246.2 1251234400 1123862291 929287845 1060333731 1021058336 CTP 31355671.93 39658126.53 30280996.72 35436149.76 43947182.09 30413588.03 28732552.78 25720471.78 25330864.52 18433918.14 20425940.58 28697366.69 dATP 3245744.745 3429464.827 3073200.228 1955609.108 2838059.874 2009190.025 8145570.96 7239708.2 7417971.335 5447330.175 5612354.772 7424747.478 dCTP 1221988.971 1306488.361 1032034.796 677946.1194 962062.3745 641308.335 1982326.97 1731895.149 1640909.104 1235891.995 1303628.718 1964657.79 dihydroxyacetonephosphate 11098713.13 8748535.572 8744466.896 6042305.751 5885170.189 5575560.702 11149503.61 11428225.3 11157547.6 12926900.01 11125326.17 10447157.59 erythrose-4-phosphate 829193.2583 796680.8243 845750.5145 984006.1756 616214.0881 831366.8604 829093.0577 622006.7339 905354.3698 843923.0667 713785.6159 735423.3666 F6P G1P 21299300.24 20441556.14 20324633.82 20731759.69 19889702.68 18859966.18 18961874 19246350.04 18042983.11 19809495.75 20028267.33 17568105.48 folic acid 468714.9438 790376.8543 538065.2989 16069.99427 751157.1218 701865.2915 908587.5987 formyl methionine 26031782.82 25816836.24 26537456.86 20514368.18 19525483.63 19420675.89 24326887.67 17790616.04 19071371.52 26484364.23 26159166.75 24755827.01 Fructose-1,6-bisphosphate 39913072.31 43020976.78 40765998.56 33213798.03 32954849 34391980.94 44757735.81 43865620.52 43989975.65 45745043.66 47258446.12 51497312.63 fumarate 88206266.68 85918236.92 93701722.5 58069884.56 60564896.31 59214354.05 84213013.62 82889817.43 79555794.65 84312715.46 84380579.61 83031779.97 GAR 14310497.68 14474068.55 13158779.25 29505992.2 27072418.87 30416636.9 15206.02419 GDP 3191726.51 3342972.878 3388511.663 2012067.555 2282782.28 2364955.046 3048058.853 3417492.068 3646976.037 3364959.866 3266488.479 3202566.253 Glucose-6-phosphate 24414771.2 23330981.76 23823796.61 26343559.24 23546910.72 24322986.26 20611850.68 20879466.34 19521237.55 20894236.27 20734491.59 18962740.14 glutamate 1957414715 1740298216 1842753505 2071216662 1864922577 2055958414 1647139969 1510510128 1598037278 1880425690 1902178052 1622813018 glutamine 8433095336 8271781963 8267180877 9290749471 9590815893 9504237648 7809530823 8234746528 8017573894 7735368317 8015080232 7950549109 glycerate 200444761.2 205324902.3 208841288.4 217196127.8 206600160.3 231070483.9 193122147.1 240534627.5 218821828.6 189576947.9 201804496 194883226.3 Glycerol-3-Phosphate 19418718.93 19065804.79 19938242.86 14127482 14226099.33 13206861.39 21670461.1 21624943.74 21017500.06 21948918.07 21888474.45 21463131.07 Glycerophosphocholine 13616364746 14107126196 14950675962 7170782848 7523021460 7250291508 18830801008 21296280148 21258616527 20429666615 20093173140 21033937151 glycine 72271692.61 63675538.74 66525511.76 45782800.8 41022971.05 45314740.75 68966143.31 68910683.41 67211072.31 41815155.83 40356111.77 35867932.51 GSH 4951583205 5004570310 4907125978 4548571403 4573489134 4703643325 4559525215 3041474556 2784605138 4225351624 5467620808 4884573289 hydroxyproline gammaALA 191093856.3 165648509.3 177241916.4 195090666.7 165763048 198638545.8 141370131.9 123043921.7 124851208.3 155394463.8 159660133.6 140107574.3 hypotaurine 3103578.559 3121016.66 3084005.619 3606345.977 4008631.639 3129365.683 3240627.735 2958079.084 2356922.25 2916640.827 3284122.009 3417136.497 hypoxanthine 1628495.528 1624939.03 1401883.865 1472591.926 1081994.408 1382656.005 1433524024 1505148022 1437190026 1434022844 1332376611 1381406215 IMP 12637778.06 11735909.13 12619867.17 7479965.721 7397888.245 8206158.227 6009806.414 6259602.495 5707452.713 6503116.696 6215874.57 5791554.702 leucine 2050808859 1920346805 2129164883 2170014668 2022864016 2122053718 2018006242 2183622236 2198737942 2200767877 1941528876 2011114274 lysine 50176783.5 56565016.34 50482955.85 63720031.04 75063454.73 60294355.69 59890433.05 63411951.29 67291891.62 59951049.19 55315713.99 58524544.67 malate 981287687.4 940692027.5 1038878261 650047155.1 662989873.5 652173620.7 862744792 898364968.5 872590109.1 900797716.3 878169565.2 915382617.2 malonyl-CoA 1258110.057 1129243.181 1269230.285 919094.653 918629.2654 809840.009 1171193.16 1424810.481 1179587.13 1216617.838 1376128.459 1167944.112 methionine 797104546.8 759918211.1 731197429.2 864634580.7 852023017.8 863411067.4 773013002.8 778698527.7 819069076.5 800411936.1 772606698 802145860.1 methylthioadenosine 372643885.8 347750693.8 370165087.1 311285636.2 305916038.8 285768955.5 381124954.3 428654105.2 491792387.4 409637876.6 341885853.6 359943738.1 N-acetylaspartate 35500162.91 30098052.52 25985359.27 37773763.65 34962836.61 33049695.04 27491540.68 23493276.33 22442135.79 24659854.24 29452087.48 23612772.25 N-acetylmethionine 7244391.114 5311058.692 7090277.65 3770858.015 3277129.755 3456740.886 8229445.659 7521520.973 9311917.015 8395244.263 7746210.629 8075289.807 N-acetylphenylalanine 1313356.475 1205458.86 1184700.417 989059.7988 1053237.3 1182496.636 1214452.436 1117073.306 1178436.852 1409909.041 1458302.792 1160618.903 NAD. 406195551.8 391893122.5 398280218.6 367723160.7 352957096.6 354903234.5 367178629 390679772.6 384855822.2 378325394 410442948.2 378145190.1 NADP 11017708.92 13540623.39 11156912.09 9811498.796 14337125.7 11814355.84 10105390.45 11980194.57 12514367.19 11479012.92 11428593.09 10248721.42 niacinamide 227230986.3 232429329.6 225298004.3 234711791.5 249871424.9 204734344.2 225314736.9 282237874.3 319609923.1 226518526.6 218231054.8 199302718.3 O-phosphoethanolamine 2115772.772 2140022.909 2261643.789 3013785.033 2959486.363 3247978.563 1743723.485 1864717.012 1745085.786 1560312.499 1816532.368 1806595.175 Pantothenate 868482303.1 853456315.7 874100624.8 766948760.6 764094560.2 761524677.1 978568359.3 1043239461 966925222.4 939131178.4 946519798.2 986242627 phenylalanine 210072463.9 196350200.2 212117302.4 234939162 213133420.6 240271569.9 202755267.5 202915595.3 204225108.9 241767418.7 207170476.1 218104604.5 phosphocholine 17777775205 16857361119 17622876623 15982022679 15346991584 14957183878 19614345124 23244848636 21833673422 18997816798 20104033245 21072199212 phosphocreatine 104469355.7 102003094.5 96254251.99 109292117.1 110135141 107008914.5 108359552.7 105802126.8 93562124.81 102789597.2 110561173.4 108484829.3 phosphoenolpyruvate 19309292.25 21050162.69 21009962.37 13421563.45 14911227.13 14978849.72 18181149.36 16742477.87 17196983.42 16309552.48 18659313.13 18891376.18 proline 10534459293 9738497765 10449748552 10647375402 10232692129 10163812712 9481128593 10025042519 9600355341 9195979021 9285640865 9766159106 propionylcarnitine 71376464.13 62640434.53 70018794.19 68445622.95 58715879.12 66148285.98 55692807.23 54450087.47 59169154.61 70391378.51 61311072.67 63616964.7 PRPP 5037562.649 5110096.089 4939400.721 30363306.85 33892030.92 31241608.71 1114315.032 833710.5723 901402.2491 856833.9854 Pyridoxine 261116370.6 284727108.4 278370084.1 243584821.4 277450088.6 233183066.5 286555961.3 270064037 286207829.4 280431881 262266726.5 255505631.9 pyruvate 17018851.39 20797135.63 19872208.68 9791812.072 10657186.96 10839103.81 15600038.99 12339135.96 7557977.974 18544760.77 19068810.96 15838338.97 quinolinic acid 640352.0593 750211.4463 580514.6276 721584.2369 807421.0179 849431.4352 586148.0285 630013.9544 719934.6001 705841.4556 689740.8482 802769.0332 Ribose-1-phosphate 649171.2796 619662.4633 646905.8249 2877906.143 2545250.973 2836572.188 298161.6353 320327.7768 304777.3782 306494.9492 316390.5387 263305.8735 Ribose.Ribulose.5.phosphate.3Me.Lys 3595944.862 3411144.762 3480276.729 3231854.814 3060312.215 3177471.577 3025222.401 3124855.701 3243149.109 3489098.031 3454863.544 3034127.058 S-Adenosyl-L-Homocysteine 7713284.43 6989234.626 8273395.02 7219944.388 7440247.789 6256299.772 7951664.362 10069769.75 9560004.656 7722575.518 8291463.083 8568367.496 Sedoheptulose-7-phosphate 5038516.464 4882547.438 4786247.304 4237614.921 4157913.704 3962112.923 5711459.335 6049801.144 5623838.819 5914587.281 6466964.561 5369502.49 serine 55915011.22 57413599.6 51398459.16 124758573 126442292.8 133024035.1 74223798.22 72782298.4 69442370.97 130407182.5 121299568.3 115657185.1 succinate 45505817.61 38678940.05 43089870.28 35203302.45 33822836.84 36664800.22 42153372.75 62137154.31 52002551.55 51424882.49 40907896.84 33881340.28 taurine 44544284.3 46032468.49 46365910.42 49303623.92 47231918.07 47627183.3 41241239.77 46005746.36 40560528.35 46555778.85 44845443.56 43674281.37 threonine 432888090.5 477550983.6 448232126 513977791.8 572993847 475284021.7 398444817.1 336141996.5 323674459.2 353015671.1 430604463.9 455703809 tryptophan 314112909.6 292402828.4 286659758.2 327063490.6 312022986.6 317586800.7 318399542 300000400.2 303884209.1 327312627 313235875.6 321330670.8 tyrosine 781150505.6 700446679.1 760145520 802033439.2 795914470.6 791325867.6 734442219.5 804779836.9 810785490.2 764580657.1 733481616.8 748542616.7 UDP.GlcNAc 187196641.9 169838073 181776612.9 139572950.3 133672965.5 133766515.4 123555268.3 129297354.6 121669281.5 133102843 133937328 127385073 UDP 4597212.317 5774758.569 5328957.777 5953542.719 5977603.189 6348854.615 2608790.154 3635394.377 4522929.213 2650713.759 2666554.828 2992438.522 UMP 1091333.249 1104025.317 1169841.615 1112514.523 1050302.231 1218879.917 574922.7092 584968.3934 633720.8725 691663.8392 714852.4061 786751.1671 Ureidosuccinic acid 3468230.923 3347421.961 3890598.444 1272605.991 1435801.524 1365094.738 20189596.22 20615039.98 20460017.94 24812720.13 24850143.06 23927937.34 UTP 246232218.5 286032110.2 231583129 302816510.8 375537780.1 259455600.5 166325939.5 149220234.7 143469271 109689611.3 126829965.3 164436201.9 xanthine 376686.0507 359220.7733 542307.4777 483484.0163 413183.0466 382757.838 xanthosine 90595.46524 61874.52697 55936.2871 68947.40225 78945.47197 119729.2188 99147.03556 58474.92676 68516.925 76422.02326 93784.36956 109186.5172 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