#METABOLOMICS WORKBENCH amaynard_20231001_202210 DATATRACK_ID:4359 STUDY_ID:ST002899 ANALYSIS_ID:AN004758 PROJECT_ID:PR001773 VERSION 1 CREATED_ON October 1, 2023, 8:29 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 erythroid progenitor cells following SHIN1 and ST:STUDY_TITLE inosine treatment ST:STUDY_SUMMARY Culture of erythroid progenitor cells cultured in SFEM II media supplemented ST:STUDY_SUMMARY with SHIN1 and/or inosine for 48 hours followed up by metabolomics targeting ST:STUDY_SUMMARY polar 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_BIOSOURCE_OR_SUPPLIER Lineage depleted erythroid progenitor cells isolated from E14.5 mice. #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 Erythroid_Progenitor EP_Vehicle_HILIC_1 Treatment:Vehicle RAW_FILE_NAME=Copy of 20220715_AM_ErythProg3_SHIN_ino_AM883.raw SUBJECT_SAMPLE_FACTORS Erythroid_Progenitor EP_Vehicle_HILIC_2 Treatment:Vehicle RAW_FILE_NAME=Copy of 20220715_AM_ErythProg3_SHIN_ino_AM884.raw SUBJECT_SAMPLE_FACTORS Erythroid_Progenitor EP_Vehicle_HILIC_3 Treatment:Vehicle RAW_FILE_NAME=Copy of 20220715_AM_ErythProg3_SHIN_ino_AM885.raw SUBJECT_SAMPLE_FACTORS Erythroid_Progenitor EP_SHIN1_HILIC_1 Treatment:SHIN1 RAW_FILE_NAME=Copy of 20220715_AM_ErythProg3_SHIN_ino_AM886.raw SUBJECT_SAMPLE_FACTORS Erythroid_Progenitor EP_SHIN1_HILIC_2 Treatment:SHIN1 RAW_FILE_NAME=Copy of 20220715_AM_ErythProg3_SHIN_ino_AM887.raw SUBJECT_SAMPLE_FACTORS Erythroid_Progenitor EP_SHIN1_HILIC_3 Treatment:SHIN1 RAW_FILE_NAME=Copy of 20220715_AM_ErythProg3_SHIN_ino_AM888.raw SUBJECT_SAMPLE_FACTORS Erythroid_Progenitor EP_Inosine_HILIC_1 Treatment:Inosine RAW_FILE_NAME=Copy of 20220715_AM_ErythProg3_SHIN_ino_AM889.raw SUBJECT_SAMPLE_FACTORS Erythroid_Progenitor EP_Inosine_HILIC_2 Treatment:Inosine RAW_FILE_NAME=Copy of 20220715_AM_ErythProg3_SHIN_ino_AM890.raw SUBJECT_SAMPLE_FACTORS Erythroid_Progenitor EP_Inosine_HILIC_3 Treatment:Inosine RAW_FILE_NAME=Copy of 20220715_AM_ErythProg3_SHIN_ino_AM891.raw SUBJECT_SAMPLE_FACTORS Erythroid_Progenitor EP_SHIN1_Inosine_HILIC_1 Treatment:SHIN1_Inosine RAW_FILE_NAME=Copy of 20220715_AM_ErythProg3_SHIN_ino_AM892.raw SUBJECT_SAMPLE_FACTORS Erythroid_Progenitor EP_SHIN1_Inosine_HILIC_2 Treatment:SHIN1_Inosine RAW_FILE_NAME=Copy of 20220715_AM_ErythProg3_SHIN_ino_AM893.raw SUBJECT_SAMPLE_FACTORS Erythroid_Progenitor EP_SHIN1_Inosine_HILIC_3 Treatment:SHIN1_Inosine RAW_FILE_NAME=Copy of 20220715_AM_ErythProg3_SHIN_ino_AM894.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 Erythroid progenitor cells were cultured for 48hr in SFEM II media containing TR:TREATMENT_SUMMARY vehicle, SHIN1 (1.25 uM), inosine (100 uM), or SHIN1+inosine. #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 EP_Vehicle_HILIC_1 EP_Vehicle_HILIC_2 EP_Vehicle_HILIC_3 EP_SHIN1_HILIC_1 EP_SHIN1_HILIC_2 EP_SHIN1_HILIC_3 EP_Inosine_HILIC_1 EP_Inosine_HILIC_2 EP_Inosine_HILIC_3 EP_SHIN1_Inosine_HILIC_1 EP_SHIN1_Inosine_HILIC_2 EP_SHIN1_Inosine_HILIC_3 Factors Treatment:Vehicle Treatment:Vehicle Treatment:Vehicle Treatment:SHIN1 Treatment:SHIN1 Treatment:SHIN1 Treatment:Inosine Treatment:Inosine Treatment:Inosine Treatment:SHIN1_Inosine Treatment:SHIN1_Inosine Treatment:SHIN1_Inosine hydroxyglutarate 786541911.2 465532202 848470436.9 1147902041 799786186.7 829919564.4 680603250.6 883204989.7 515762343.1 1204592962 990573403.8 850412958.8 2-oxobutyric acid 44822451.4 52716116.88 45619113.89 63478080.22 50111881.29 58321549.51 42343339.35 42072279.69 40849678.42 48769060.76 39012793.03 45135088.27 3-methyl-2-oxopentanoic acid 4240516.062 5418690.65 6977386.066 3243883.333 4468840.324 4358598.062 5998255.995 5085570.93 5739270.369 6535394.824 5492833.863 6754701.511 3-phosphoglycerate 5165927.639 4475660.302 4434036.384 8751791.741 5649378.658 3999661.562 3966338.444 4945527.608 4203189.635 2826300.308 3437079.848 3777568.214 6-phosphogluconate 214692.9593 193053.1775 272091.0899 734895.8667 378564.8338 383735.7455 330585.3168 352731.3959 253842.6537 158724.811 181700.4967 265176.1862 Acetyl-CoA 2156149.193 2409363.551 1641313.68 1064567.172 674692.0878 640728.6822 1746707.277 2043861.932 1685525.793 1833161.655 1965196.247 1884565.968 AICAR 0 47496.68745 0 4875473.64 3557129.55 2091818.763 28790.13882 30531.33572 23580.63246 0 0 37079.46159 alanine 29313550.26 25745270.61 27133512.09 32710144.44 34841093.17 38669048.62 27085705.92 23395031.72 25271430.61 29715010.89 25140793.06 25989127.87 AMP 778836.1044 1065014.302 808842.5746 251776.2645 154179.53 205829.871 820932.9739 598362.5806 454381.0487 823274.3744 420674.4664 831592.8603 arginine 391988905 425429960 494232831.3 609119975 727584383.6 742699472.5 437609082.7 369379709.4 462055885.2 423876453.5 389945277.4 394545209.5 argininosuccinate 2410972.59 2794901.307 2155953.072 2327468.641 1805672.865 1559807.308 1746255.042 2133228.753 2010735.396 1782053.981 2110234.504 1912775.47 ascorbate 85631559776 94234254951 98975987996 1.23E+11 1.10E+11 1.12E+11 81404478092 90376564637 80530261855 97594143679 91095857098 94992110771 asparagine 102711776.9 98200872.58 98949832.98 85351396.57 93419519.77 80610946.88 89895356.86 99429281.25 88222414.94 84813610.23 84394663.86 96746637.65 aspartate 85675984.47 84548820.75 75663014.36 81752550.64 67994592.86 44626894.62 64572404.9 80247868.92 61059804.56 37562959.64 46626910.58 46904333.57 ATP 77942356.64 71642437.2 73655559.7 41449899.88 31246436.66 26240856.16 68106037.75 81651749.47 67337983.71 48908499.66 63744844.84 70738255.34 beta-alanine 15881043.48 12912656.77 10868650.96 16044799.59 15567339.49 10535766.74 11174316.58 14656982.54 9075956.57 15818644.54 17990188.1 20837697.49 betaine 69442831.23 79845243.67 56252624 61030285.31 48554495.47 44662441.85 49822664.55 57313891.08 44138642.82 126078741.6 38254065.35 53486668.39 carnitine 11717280.74 11247093.85 10648065.51 14713992.12 14580844.47 11687868.39 8866016.882 10515144.82 8215374.829 15317272.26 9178132.541 9934803.34 citrate 452043827.4 416590745.9 403022862.4 421717842 345504548.3 293747077.8 315146208.7 408795229.8 305190616.1 320100851.8 326881066.3 339391041.7 citrulline 3594052.398 4754274.832 2025301.68 1815452.85 2951348.339 23464337.94 26445517.05 1591254.96 2180418.689 3044055.314 3726805.385 1547168.62 creatine 24764850.27 19581300.54 18032450.88 30768336.54 23485964.02 36135073.67 18256304.1 19177248.03 16100775.51 27414316.4 20164200.68 17777539.79 creatinine 3045685.772 1735030.182 2907328.503 2537954.151 2085813.029 7089206.28 2605474.442 1997526.484 1828573.259 6237734.159 1723191.19 1738592.693 CTP 8224262.229 7645896.796 7412838.455 5665106.459 3835250.978 2381133.917 5526637.852 7425059.206 5497438.985 4189637.059 5298162.881 5581507.346 cystine 7586974.647 9540351.856 13669657.83 21150872.99 31678672.5 36817731.88 13426753.86 8656571.945 13759508.89 13196632.21 11575738.73 12613855.84 dihydroxyacetone phosphate 1872255.149 1407974.401 1335965.204 402205.83 NA NA 1150071.14 1954219.296 1154193.949 1695642.676 1812011.376 1229035.423 F6P_G1P 5575604.876 6076182.299 5974875.935 6101800.759 4767938.911 3585456.639 4809253.198 6419742.336 5309015.226 4709757.515 5291475.373 6271309.241 folic acid 1083541.701 1389726.936 1523642.006 1235330.199 1947072.074 2294209.913 1460618.648 980189.2226 1612714.494 1510929.977 1145934.957 1310556.495 Fructose-1,6-bisphosphate 4811904.333 5196981.197 5192189.287 1887848.315 1113934.226 799466.5461 2921809.914 4892581.871 3919100.282 3305443.321 4375945.94 3874766.788 Glucose-6-phosphate 2520266.811 2899646.81 3224900.728 5548428.546 5146696.602 4636584.785 3038124.623 3919099.864 4084381.153 3083143.589 3890235.318 4600699.418 glucose 8348118.818 8956276.157 11628837.03 15209477.32 21603183.44 21534196.22 10647312.45 8013548.76 8410919.05 8119833.625 7114579.591 10154190.59 glutamate 171749903.1 166395785.7 166965333 116149073.5 99016036.43 75542234.49 137254691.3 166432669.7 136262902.2 104590519.2 125048512.1 126678130.8 glutamine 3722725428 3814384770 3937887920 4194080994 4732323002 4560396445 3646231132 3789122288 3507589167 4058474898 3481039343 3793802227 glycerate 154792110.5 167727025.7 150051333 218903944.9 190596156.7 200683731.1 147776971.1 150360050.6 156717405 182305110.5 156639879.6 161867219.1 Glycerophosphocholine 1595981909 1362438624 1249975804 129920039.6 129954883.8 127230194.2 1297987965 1489045642 1631411880 1507521094 1343583021 1283464224 glycine 32190583.25 30694512.09 28775565.99 25476587.05 23184188.81 23614258.25 24835006.88 29460464.74 27959506.86 29181592.14 34912608.54 32861435.66 GMP 149516.0463 197890.571 36758.9923 110460.8331 38836.0464 NA 94267.32225 28868.60768 109538.1461 28346.19429 65377.24592 71191.07338 GSH 2517844921 2310507654 1966735232 2070786150 1433131980 1078811617 2114651333 2405489367 2090826816 1571875800 2658313456 2116281325 GSSG 34628956.64 23612921.77 26596265.63 28303314.78 20485241.8 16911055.87 25824974.91 33512761.74 36956645.1 37972834.21 25379380.63 26697237.73 GTP 5770083.723 5141807.699 5125775.011 3834131.19 2747334.907 1950191.834 4515685.503 5181823.141 4173370.274 3233423.125 3921385.74 4580944.854 guanidinoacetate 5433233.864 4895444.368 4488939.382 5948089.796 4685725.079 4002098.569 4977338.564 4860725.627 5073001.475 4142776.86 4914433.248 2944583.987 histidine 182590787.3 193448735.2 203553702.1 309551704 378808814.5 412844406 248350658.6 169733930.3 188294584.3 220452447.5 192935313.5 185159980 hydroxyproline gammaALA 3643219.037 3508175.157 4171526.901 5301765.583 4487974.906 5135443.805 2889766.235 2414236.205 1499484.308 3639810.302 1897478.333 3913984.057 hypotaurine 1265952.384 1106433.991 852127.7992 330021.8493 320119.9944 NA 695148.0931 1221834.992 346142.9557 834523.4399 1177392.69 1117234.222 hypoxanthine 10922153.99 10566598.98 6358404.954 6160179.308 4779015.245 5372916.064 249197144.7 233412301.7 242089672 222285229.1 272487547.8 244733966.6 IMP 971293.2293 848349.2017 762751.715 1352293.954 873943.3723 589046.7302 851127.5875 1166791.78 895149.0013 669091.1361 851626.9491 777949.8707 isoleucine 278695969.9 271144642.9 292002178.5 345664245.1 403966258 430338026 288398118.8 261981360.4 298634617.6 306035158.1 295745811.2 282906935.3 leucine 257969804.8 271068113.3 291650967.9 348901892.1 405231818.7 441881728.5 297464981.9 236179120.4 270079393.6 270547190.2 266296121.1 285411762.5 lysine 75456891.79 80903986.18 99468747.2 124531226.2 154606490.2 167242138.9 90883298.12 75067361.42 101577697.5 87898763.62 82120691.29 85006979.3 malate 154242658.4 161360484.5 132852471.4 78046553.29 65354865.45 59174845.67 116700858.3 143430621.9 111164790.7 136509270.7 110064407.7 132454259.9 Methionine sulfoxide 30143111.57 25708732.71 34718363.49 35607242.59 42378594.8 51726737.19 32211132.8 26839598.41 32783831.2 39412394.47 34503579.74 31622689.03 methionine 88071259.49 88907691.12 91150553.37 110018298.4 135575955.9 136319716.7 92922268.62 86951944.87 93419710.92 107513714.3 102032593.2 93340686.64 methylthioadenosine 3817583.863 4073969.734 3685552.834 3196872.826 1935876.352 1377216.507 2996820.328 2855378.692 3420652.011 2740615.971 3977759.526 3019755.084 NAD 35126451.34 32813592.65 32338643.6 31713385.73 21094723.19 15005892.81 29885946.28 34060745.73 31451783.88 22907405.55 34763768.85 31265025.81 niacinamide 113327561.1 129053894.3 144631097.6 163945817.7 194115114.8 210332123.2 155873129.5 106613095.2 184818931.9 129897941.7 138616603.3 138021988 ornithine 6246504.521 6998090.675 6378609.875 4050610.705 4228237.504 9591486.949 6998075.095 5396977.155 7485664.177 5273037.892 6714503.176 5296422.83 orotate 714247.306 783740.8166 518525.7322 NA NA 216788.9249 2178582.289 1377044.56 1790688.573 964812.9598 2001542.114 2053109.383 Pantothenate 127520741.4 153042363.3 143211746.3 179631994.7 195512080.4 161679298.2 120249886.2 136792993.8 156083320.4 141429086.2 154419058.6 169573156 phenylalanine 51245075.77 61035509.18 59038156.74 63706940.78 76455385.86 83296795.44 57533854.82 54028403.21 58196680.4 53503923.87 51692757.34 57538238.71 phosphocholine 1565241513 1501721867 1234919696 265047793.8 197939114.1 158460549.9 1287251964 1389990979 1509351713 1304362433 1435713967 1203349256 phosphocreatine 182446.1191 150274.2914 141755.4358 55531.71025 NA 55356.3704 219566.2103 221970.2726 170292.3889 46228.63668 92564.76153 79042.14086 phosphoenolpyruvate 2688748.625 2562590.407 2024909.446 5241274.115 3600639.247 1927771.55 2149694.697 2547960.733 1692524.158 1620675.977 1985844.555 2104447.362 proline 1484976680 1399209086 1433966386 1144637642 1142144225 1038847917 1398929004 1591727671 1370237580 1555904657 1792266412 1818108551 PteGlu 1083541.701 1375393.786 1523645.006 1224918.185 1947069.569 2264278.091 1460622.47 947076.4892 1612720.738 1510937.074 1128509.703 1299851.783 Pyridoxine NA NA NA NA NA NA NA NA NA NA NA NA Ribose_Ribulose-5-phosphate-3MeLys 476361.1162 443126.0119 662423.0801 571710.5693 444597.8844 398532.7989 458128.8166 588572.227 400204.7694 646222.4254 559880.4124 387123.7614 S-Adenosyl-L-Homocysteine 2469028.093 1064752.202 2240542.852 418696.8311 NA NA 1878259.103 2091180.905 2112602.951 2197164.762 2986525.397 1708590.24 sarcosine NA NA NA NA NA NA NA NA 1103737.458 NA 1215449.774 NA Sedoheptulose-7-phosphate 1496362.803 1708459.249 2023882.739 1253382.713 1159855.052 641580.887 1951424.272 2375220.181 2157332.687 2443630.234 2099621.989 2545479.1 serine 24333220.68 24586849.14 25314196.96 32495693.82 34440787.26 43599950.97 25711532.84 21738162.79 23447672.54 32655594.04 27761742.4 29745764.38 succinate 29744320.67 25708421.2 29675579.33 33247152.15 26988140.19 35122481.18 33790229.88 32248217.51 22474571.69 36182141.77 30963897.69 32715169.03 taurine 14198100.97 15734218.52 16564873.58 1107566.224 1373815.227 1449344.337 13044759.03 17129570.31 10786661.35 8614708.141 12854150.11 10115568.23 threonine 239787420.6 206291566.6 195139445.4 210885904.7 255076668.1 313270005.8 196118302.1 183702324 219405564.2 200763176.7 238129235.4 205824179.2 tryptophan 50542907.2 50711064.89 52396609.15 57390469.24 63991628.97 69171079.74 52896906.78 49283395.04 49637234.09 57941187.81 52992823.81 53551525.17 tyrosine 207166455.4 195170037.4 239551530.3 263547311.6 306045314.3 308483729.3 249089312.3 223033303.2 223844952.8 251206977.8 244216510.2 244580485.7 UDP-GlcNAc 20281274.97 19638465.57 16856570.36 8555910.284 6487343.773 5310757.007 14525758.81 18500033.8 15136126.39 12726369.71 17473868.35 16490134.91 UDP 1866201.173 3025459.143 1735235.328 812501.2024 736529.0388 136598.13 773210.1341 1280544.329 793237.7409 506265.5058 558452.6126 898946.48 uric.acid NA NA NA NA NA 65119.3475 NA NA 16777.89991 1291773.076 NA NA UTP 20985375.74 20009423.83 19626157.26 18411447.03 12715715.73 8662049.946 13645022.6 17102916.06 14689057.96 10401986.51 12496302.95 13722415.67 valine 93522130.58 86635989.55 96965722.61 116982198.9 156009142.2 150135141.9 94947349.15 119298246.2 103057273.1 70396360.19 100233879.4 115179510.2 MS_METABOLITE_DATA_END #METABOLITES METABOLITES_START metabolite_name Standardized name Formula Exact mass Sub class hydroxyglutarate 2-Hydroxyglutaric acid C5H8O5 148.0372 Dicarboxylic acids 2-oxobutyric acid 3-Methyl pyruvic acid C4H6O3 102.0317 Oxo FA 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 AICAR AICAR C9H14N4O5 258.0964 Imidazole ribonucleosides and ribonucleotides alanine Alanine C3H7NO2 89.0477 Amino acids AMP AMP C10H14N5O7P 347.0631 Purine rNMP arginine Arginine C6H14N4O2 174.1117 Amino acids argininosuccinate Argininosuccinic acid C10H18N4O6 290.1226 Amino acids ascorbate Vitamin C C6H8O6 176.0321 Gamma butyrolactones 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 betaine Betaine C5H11NO2 117.0790 Amino acids carnitine Carnitine C7H15NO3 161.1052 Carnitines citrate Citric acid C6H8O7 192.0270 TCA acids citrulline Citrulline C6H13N3O3 175.0957 Amino acids creatine Creatine C4H9N3O2 131.0695 Amino acids creatinine Creatinine C4H7N3O 113.0589 Imidazolines CTP CTP C9H16N3O14P3 482.9845 Pyrimidine rNTP cystine Cystine C6H12N2O4S2 240.0239 Amino acids dihydroxyacetone phosphate Dihydroxyacetone phosphate C3H7O6P 169.9980 Short-chain acids F6P_G1P - - - - folic acid Folic acid C19H19N7O6 441.1397 Folic acids Fructose-1,6-bisphosphate Fructose 1,6-bisphosphate C6H14O12P2 339.9961 Monosaccharide phosphates Glucose-6-phosphate Glucose 6-phosphate C6H13O9P 260.0297 Monosaccharide phosphates glucose Glucose C6H12O6 180.0634 Hexoses glutamate Glutamic acid C5H9NO4 147.0532 Amino acids glutamine Glutamine C5H10N2O3 146.0691 Amino acids glycerate Glyceric acid C3H6O4 106.0266 Sugar acids Glycerophosphocholine sn-Glycero-3-phosphocholine C8H20NO6P 257.1028 Organic phosphoric acids glycine Glycine C2H5NO2 75.0320 Amino acids GMP GMP C10H14N5O8P 363.0580 Purine rNMP GSH Glutathione C10H17N3O6S 307.0838 Tripeptides GSSG Oxidized glutathione C20H32N6O12S2 612.1520 Tripeptides GTP GTP C10H16N5O14P3 522.9907 Purine rNTP guanidinoacetate Guanidoacetic acid C3H7N3O2 117.0538 Amino acids histidine Histidine C6H9N3O2 155.0695 Amino acids hydroxyproline gammaALA - - - - hypotaurine Hypotaurine C2H7NO2S 109.0198 Sulfinic acids hypoxanthine Hypoxanthine C5H4N4O 136.0385 Hypoxanthines IMP IMP C10H13N4O8P 348.0471 Purine rNMP isoleucine Isoleucine C6H13NO2 131.0946 Amino acids leucine Leucine C6H13NO2 131.0946 Amino acids lysine Lysine C6H14N2O2 146.1055 Amino acids malate D-Malic acid C4H6O5 134.0215 Beta hydroxy acids Methionine sulfoxide Methionine sulfoxide C5H11NO3S 165.0460 Amino acids methionine Methionine C5H11NO2S 149.0511 Amino acids methylthioadenosine Methylthioadenosine C11H15N5O3S 297.0896 Purine ribonucleosides NAD NAD+ C21H27N7O14P2 663.1091 Nicotinamide dinucleotides niacinamide Niacinamide C6H6N2O 122.0480 Pyridine alkaloids ornithine Ornithine C5H12N2O2 132.0899 Amino acids orotate Orotic acid C5H4N2O4 156.0171 Pyrimidine carboxylic acids 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 PteGlu Folic acid C19H19N7O6 441.1397 Folic acids Pyridoxine Pyridoxine C8H11NO3 169.0739 Pyridine alkaloids Ribose_Ribulose-5-phosphate-3MeLys - - - - S-Adenosyl-L-Homocysteine S-Adenosylhomocysteine C14H20N6O5S 384.1216 Other purines sarcosine Sarcosine C3H7NO2 89.0477 Amino acids 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 uric.acid Uric acid C5H4N4O3 168.0283 Xanthines UTP UTP C9H15N2O15P3 483.9685 Pyrimidine rNTP valine Valine C5H11NO2 117.0790 Amino acids METABOLITES_END #END