#METABOLOMICS WORKBENCH amaynard_20230822_102535 DATATRACK_ID:4239 STUDY_ID:ST002831 ANALYSIS_ID:AN004624 PROJECT_ID:PR001773 VERSION 1 CREATED_ON August 25, 2023, 1:36 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 Folate depletion upregulates heme synthesis ST:STUDY_SUMMARY Culture of K562 cells in RPMI media containing 2,000 nM folic acid or 100 nM ST:STUDY_SUMMARY folic acid for 6 days followed my polar metabolomics. 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 2 #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_1 Folic_Acid_Concentration:2000_nM RAW_FILE_NAME=20210201_QEF1_HILIC_AdamHeme6KO_AM304.raw SUBJECT_SAMPLE_FACTORS K562 K562_2000nM_2 Folic_Acid_Concentration:2000_nM RAW_FILE_NAME=20210201_QEF1_HILIC_AdamHeme6KO_AM305.raw SUBJECT_SAMPLE_FACTORS K562 K562_2000nM_3 Folic_Acid_Concentration:2000_nM RAW_FILE_NAME=20210201_QEF1_HILIC_AdamHeme6KO_AM306.raw SUBJECT_SAMPLE_FACTORS K562 K562_2000nM_4 Folic_Acid_Concentration:2000_nM RAW_FILE_NAME=20210201_QEF1_HILIC_AdamHeme6KO_AM307.raw SUBJECT_SAMPLE_FACTORS K562 K562_100nM_1 Folic_Acid_Concentration:100_nM RAW_FILE_NAME=20210201_QEF1_HILIC_AdamHeme6KO_AM308.raw SUBJECT_SAMPLE_FACTORS K562 K562_100nM_2 Folic_Acid_Concentration:100_nM RAW_FILE_NAME=20210201_QEF1_HILIC_AdamHeme6KO_AM309.raw SUBJECT_SAMPLE_FACTORS K562 K562_100nM_3 Folic_Acid_Concentration:100_nM RAW_FILE_NAME=20210201_QEF1_HILIC_AdamHeme6KO_AM310.raw SUBJECT_SAMPLE_FACTORS K562 K562_100nM_4 Folic_Acid_Concentration:100_nM RAW_FILE_NAME=20210201_QEF1_HILIC_AdamHeme6KO_AM311.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 6 days in either 2,000 nM folic acid or 100 nM TR:TREATMENT_SUMMARY folic acid containing RPMI media. #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 Arbitrary Units MS_METABOLITE_DATA_START Samples K562_2000nM_1 K562_2000nM_2 K562_2000nM_3 K562_2000nM_4 K562_100nM_1 K562_100nM_2 K562_100nM_3 K562_100nM_4 Factors Folic_Acid_Concentration:2000_nM Folic_Acid_Concentration:2000_nM Folic_Acid_Concentration:2000_nM Folic_Acid_Concentration:2000_nM Folic_Acid_Concentration:100_nM Folic_Acid_Concentration:100_nM Folic_Acid_Concentration:100_nM Folic_Acid_Concentration:100_nM 2,3-Bisphosphoglycerate 14515514.32 12088118.91 11465938.19 13352804.04 10516803.06 12182877.81 11724027.61 12347832.37 2-hydroxybutyric acid 2932280.017 2784813.901 2629777.612 3127329.089 3982942.674 3406474.829 3434585.829 4015758.783 2-hydroxyglutarate 158007471.1 155112791.9 183123627.9 145713662.8 224222722 247155203.8 236689823.8 227914190.3 3-methyl-2-oxopentanoic acid 15351842.41 17076892.79 17205432.95 21928987.48 8445858.052 5849933.336 3307794.441 9546537.766 3-phosphoglycerate 114550922.9 106332037.9 101404251.2 113155846 113404485.3 86759677.5 89220358.81 107511602.4 3-ureidopropionate 1384065.372 1177037.927 1739581.424 1251824.155 1391026.731 1574650.641 2064207.362 2370753.03 acetoacetate 4287562.96 4494242.694 4089487.959 4770335.949 7073757.063 5079437.959 7706403.756 6923618.578 Acetyl CoA 17294473.81 15505808.67 15809367.37 15079360.91 12113093.89 11586707.16 10262207.57 11417738.2 acetylalanine 16191936.67 16293152.25 11921288.51 12182045.05 17308426.88 16071418.75 15803492.22 17509501.88 acetylaspartate 51813805.63 50120462.57 53551381.12 41258384.38 118167913.8 119179554.9 126726408 119370320.9 acetylcarnitine 39900866.5 39910290.63 40739272.19 42444788.26 29501225.78 25856734.26 34344410.85 28549927.36 acetylglutamate 7215393.787 6900550.004 7379338.885 7370315.311 1517746.157 1403499.778 1538619.994 1806116.006 acetylserine 32200885.17 27203631 28845014.33 28849614.34 4774268.431 5686855.318 6118822.402 4959942.654 ADP 41911585.71 43539911.74 43212323.8 41309914.2 26762203.68 27455992.74 28742655.61 27487466.05 AICAR 28492321.52 24930208.84 29189001.49 27212774.69 134485425.5 141909913.7 143321366.5 132495971.7 alanine 17738185.74 18235118.78 19777980.96 16492207.76 36454948.07 40615490.21 39572416.43 32738865.61 AMP 8274053.643 8159298.582 8498969.824 8038866.716 7774215.321 8184352.707 8752280.587 7586057.971 arginine 4613622450 4942361043 4789294300 5313876826 6455657497 6568745727 5768827465 6245380402 argininosuccinate unc 40950001.29 44953064.14 46619594.85 35456951.09 43911421.97 32872686.34 35259983.1 41582642.31 asparagine 266100307.2 266479015.2 288797433.9 266261847.1 294162742 318311840.7 294588544.5 278445428.3 aspartate 535547519 559183869.5 579600601.7 528296356.4 840585971.4 751895947.8 758648860 788288925.1 asymmetric dimethylarginine 14440717.02 14251340.53 15039320.76 14685690.69 15327320.41 14114059.07 14261263.46 14251043.71 ATP 465706874.5 525617803.8 558096177.6 541496173.2 211014263 238160290.2 211821655.9 278225592.8 beta-alanine 71438935.97 66961031.37 72177332.7 65188324.15 81401967.33 86569996.09 88954579.61 86252455.66 betaine 710643429.1 645238795.6 708061297 689731214.6 610714577.2 573705256.3 782557522.4 605237242.1 biotin 449927652.3 413214414 425760262.1 419827856.7 463345298.4 477543881.7 457773497.5 446704830.3 Butyrylcarnitine 8358109.441 7647830.74 7286489.122 7032393.732 2640638.017 2276508.257 4387943.24 1947800.025 C5 Carnitines 67304325.37 65330674.13 68806974.27 67310323.48 75918814.77 70454373.57 79792061.26 76388557.04 cAMP 3859693.283 3633672.294 3401221.415 3455677.794 4782036.057 4618281.303 4630172.759 4373157.38 carnitine 121037929 110264337.3 117809526.6 112378692.3 95378129.57 103937278.2 124904413.3 99664677.49 CDP 2073721.066 2716497.931 2814856.046 2529432.794 1217805.179 1329789.186 1083403.427 1637031.281 choline 301908835.9 278469166.1 277981822.1 273831568.2 117877405.8 105657883.4 102553266.2 104218732.5 cis-aconitate 22401771.18 21838889.1 22931544.35 22711324.42 27628474.91 24554206.71 25730534.89 26409410 citrate 758250526.7 684076486.8 732070844.4 724749950.4 670373839.8 616627243 626320054.6 671909268.4 citrulline 3557213.722 2892466.469 3595603.677 3829046.589 4301643.008 5446772.489 5144151.109 4507764.159 CMP 243724.698 262249.935 306861.9214 283640.89 243232.3494 151767.4153 198268.1448 178245.8589 Coenzyme A 11608919.5 9807486.9 8741630.821 9732320.666 11251977.73 10305530.38 9625070.485 10103259.58 creatine 443159688.3 428323618.2 428646950.2 406931148.6 1255758624 1297333890 1248278498 1128764767 creatinine 0 1905566.862 2420974.048 2649890.871 4026442.545 3835858.679 4620198.082 0 CTP 36250069.9 42066219.73 43335457.35 44603843.33 21195573.22 23350945.6 20629836.44 28192475.01 cystathionine 86365887.24 59510082.79 70628816.5 45332773.24 41703199.29 50611864.63 51683882.54 42673065.27 cystine 33601698.13 48981584.97 38400686.73 72129617.21 67298218.71 78990568.9 53498198.8 69752634.16 cytidine 8446432.359 12082028.02 8290099.228 10305483.18 0 4386999.405 8148555.435 2840583.97 dATP 4067178.656 5057999.303 5149522.233 5020519.82 1852127.695 2833528.654 2337029.827 2197248.303 dCTP 1112429.373 1432730.98 1538693.917 1474693.306 935600.4252 577045.7487 513742.2628 1073709.094 dihydroxyacetone phosphate 43813097.05 32183455.05 30353695.72 34313533.51 30997538.72 28966632.68 21564233.82 32998788.48 dimethylglycine 19314069.26 16987497.83 12973537.64 14880854.55 25038621.28 25866830.89 23992785.31 23977492.24 dTMP 660406.5169 810973.3473 695865.2476 800940.8775 208839.954 243968.4343 199475.2803 95728.12454 dUMP 301871.985 291398.4206 338926.2491 403607.8683 17789498.61 4935532.393 5100601.482 13729061.7 erythrose-4-phosphate 1383427.618 1348679.994 1325500.806 1197641.33 655060.9964 905354.8389 701722.3528 512370.1876 F6P_G1P 23738795.64 21616522.56 23111885.51 22683593.33 16717086.1 20580922.44 19243051.33 16588746.64 FAD 1359607.399 1619441.945 1245237.344 124841.4931 3063799.924 2711896.593 1392822.194 2525829.367 folic acid 791697.3673 1222759.476 924737.5574 1413475.842 0 0 0 0 formyl-methionine 7355497.515 6246302.027 6922752.662 7907877.587 4262491.081 4203986.675 3699995.442 5744519.927 Fructose-1,6-bisphosphate 72002752.91 73509892.33 73776139.64 72734637.11 46252908.61 49990337.25 49427465.2 50735684.11 fumarate 53600384.42 51882469.5 61736650.71 49046165.98 45686781.24 38049601.71 39156228.7 43626337.23 GABA 325948562 305226511.9 337615639.9 288662620.7 250893763.4 271137201.4 267045572.2 240887291 GAR 19218374.95 17019981.6 19583359.9 18021674.63 66834995.59 60846675.52 63434476.97 65999046.15 GDP 3827321.726 3911579.264 3793284.123 3978452.825 2393993.283 2618148.761 2891261.017 2528803.6 Glucose-6-phosphate 22637344.9 21803250.44 23256774.08 22451395.96 12229104.74 16013096.05 14539799.8 12051237.4 glucose 10803151.31 11109037.01 11397189.56 11456177.39 16722549.13 21499925.19 19955406.03 21231045.91 glutamate 1897977894 1919473124 2071673797 1858815436 2469576184 2027133575 2092284217 2381171749 glutamine 7786388030 7507367012 8058709250 7463425082 9437301709 10117253252 9477590635 8965110160 glyceraldehyde-3-phosphate 405654.9085 302290.2302 374793.5942 350249.7691 0 0 0 0 glycerate 4113868393 4274891969 4902607930 4101029465 5869546173 6528124627 6013660729 5983222773 Glycerol-3-Phosphate 15831489.88 14862681.75 16761906.44 16080445.15 16006047.7 15387387.37 15607792.92 16524724.83 Glycerophosphocholine 28372149557 28046534316 28483339612 28084056379 12107944799 11556570818 11047661023 11140152975 glycine 27690014.7 29394363.53 30441335.85 26490436.36 29834474.72 31904499.11 32604051.67 30424042.76 GMP 355431.2644 414402.681 403485.3755 331936.7535 268610.6795 316389.2731 275895.8121 271879.065 Glutathione 4584540429 4897659624 5470056356 5316521360 5244865323 6084190260 5386418210 6188208183 Glutathione disulfide 540532472.2 418860044.9 369805165.5 304012996.2 331811091.9 331464975.5 466319474.5 245471818.8 GTP 31086299.1 37941912.43 37367589.88 41814791.69 14728284.98 18722218.48 16310896.67 19161921.17 guanine 4100026.014 3940276.782 3195470.948 3238244.541 472875.0029 0 665965.0031 389627.263 guanosine 2339402.01 2525794.023 2767575.459 2544454.912 291392.5034 249528.6382 55011.09093 265241.4813 histidine 744233995.7 764658986.4 781788111.6 756132730.6 1496292993 1574854021 1398643877 1372999376 hydroxyproline 82689557.33 77836654.07 89453557.7 79538885.05 90845039.92 92702483.47 92277812.46 92380698.48 hypotaurine 639952.111 249486.6094 0 217326.4924 2832192.741 1524546.398 388070.8829 3213551.118 hypoxanthine 4315375.681 4263422.39 4353137.297 4152664.906 0 905972.9696 939356.6062 881164.0381 IMP 5713399.219 4824089.869 5025835.665 5052832.72 5373114.121 6042027.831 5619734.399 5382990.757 inosine 3053806.388 2661805.883 2611639.178 2973284.991 0 0 474183.6308 0 isoleucine 1388211071 1483753218 1257933533 1470556660 1277098369 1353512723 1247746600 1221881021 Itaconic Acid 23701682.17 19657310.27 25868998.74 19915233.69 34786578.28 43294836.87 42528060.35 38961747.75 kynurenine 3607512.81 4063936.825 3796419.962 5607627.948 4328453.139 2774908.77 2609923.961 2259569.369 lactic acid 474582712.6 460167073.6 454244595.1 452865252.2 247729623.6 161561918 213802469.7 232324055.6 leucine 1492773065 1561977764 1439826681 1530736769 1401003420 1443949188 1374661399 1285086958 lysine 144855657.3 159222464.4 141489296.6 214693419.9 185503063 193574799.5 164064143.9 194021128 malate 528102256.1 515089029.8 573542049.4 486665580 469602049.2 408458131.1 412396708.3 446002581.9 Methionine sulfoxide 569900928.1 756288013.9 634960460.6 693385045.5 390506723.7 666231611.2 791536525.3 355444142.7 methionine 486906988 467430337.9 378127703 435215522 505264540.4 439634769.5 353885920.7 499851325.8 methylthioadenosine 62527230.76 56656008.71 45480017.71 48257486.57 42896930.96 41179837.53 36552275.77 36381320.95 myo-inositol 492287327.7 466871766.5 509769463.7 481260889 944364665.4 940521714.2 958382474.4 924482906.1 N-acetylglutamate 1084474.817 1059519.501 2097104.857 1211526.446 2082492.817 3575725.237 4918409.74 2764003.675 N-acetylglutamine 10557739.68 9575930.954 9829757.777 9706003.268 4618275.186 4751897.074 5244726.999 4812099.299 N-acetyllysine 2498969.311 2077480.449 1697924.192 2930771.108 1995345.663 2995449.162 3457483.462 0 NAD+ 491343965.5 480896810.5 523407993.1 482630248.7 352089908.7 357434676.2 342269103.3 347000919.9 NADH 13577380.95 14792824.03 13608411.12 15916563.06 6353924.987 2143375.4 2577606.737 5084733.637 NADP 12484501.66 12455703.62 11024821.69 10946515.24 12876872.45 16556208.51 18288513.49 13352575.92 NADPH 5243118.889 4857246.331 6695138.249 7186352.422 4112162.277 2154238.749 700076.7699 5264214.395 niacinamide 6850994.733 6402808.491 7313117.575 6888870.899 8588303.928 13522771.06 13172385.06 11363329.59 Nicotinic Acid 4817733.838 6692434.045 6373336.576 7063502.015 7324126.521 7709812.631 6913043.095 7605506.257 O-phosphoethanolamine 6018714.472 5772408.25 6654361.859 6140118.202 57657303.73 55004476.25 58854066.58 56370324.19 ornithine 40728367.63 41726397.72 38966567.54 40987241.6 47711285.42 47927754.2 44114969.98 47331488.52 orotate 4875491.834 4394586.347 4700963.089 3870282.013 2313618.354 2616361.234 3032775.524 2347116.286 palmitate 81047945.9 81062631.83 49337700.71 77840172.24 73073538.79 56050221.03 68832791.24 72302189.86 phenylalanine 210120234.6 219206073.6 237908750.4 226010804 193847088.1 201000098.2 181084071.8 184948248.5 phosphocholine 36562847960 33980791203 36122982296 32423021172 19307273846 19499035495 19847632681 18911079202 phosphocreatine 14563994.7 12797803.97 16103811.61 13266339.45 44894052.87 45854505.42 45512861.02 45868857.82 phosphoenolpyruvate 27318899.11 25500878.41 26757988.36 29549653.03 26026256.92 33821415.82 32391906.39 37953208.54 phosphoethanolamine 5480644.093 4625052.454 4547560.407 5078199.058 50190769.29 51152494.17 51556855.07 50632356.39 phosphoserine 21102645.81 21813745.77 23058978.43 22178401.78 13668764.51 12909638.96 13656363.5 13305128.77 Pipecolate 2444591.212 2854677.255 3073850.85 2879431.676 4577634.538 4629461.146 4948385.768 6525799.081 proline 9186896011 8827983595 9446539050 8688655649 8704990286 9079750136 8809937169 8808110819 propionylcarnitine 17423982.17 15436835.04 18636377.15 16611907.83 10281729.49 11236956.66 11764217.21 10088112.77 PRPP 1900644.24 1739656.782 1523882.992 1829694.815 17233176.37 21530995.17 22671134.93 17983799.69 Pyridoxal 30103026.01 20522603.62 16194482.84 15960144.55 33968893.39 28095723.77 22378669.46 32607698.91 Pyridoxine 238923848.5 241031613.1 232679326 289464540.2 305192573.2 310103372 250533176.7 274967916.1 pyruvate 13482462.21 14546124.97 16442362.9 15174070.09 22986146.64 21165715.62 25737586.58 27573193.69 Ribose 1-phosphate 5526162.037 4900490.376 4814134.015 5151959.123 6389759.399 5533995.773 5094924.151 5967333.797 Ribose_Ribulose 5-phosphate_3Me-Lys 5526162.037 4900490.376 4814134.015 5151959.123 6389759.399 5114512.415 4770589.742 5967333.797 S-Adenosyl-L-Homocysteine 35636580.57 32076232.78 28340404.93 28406146.48 27644264.12 27439957 26186248.92 23710576.2 S-Adenosyl-L-methionine 264186353.6 271302663.5 294022191.1 245156800.3 440402637 377668402.3 413163884.4 406376528.6 sarcosine 2088988.941 1693420.379 1943691.702 1793050.991 3899148.064 2944382.062 3839472.293 3130477.955 Sedoheptulose-7-phosphate 9092715.739 8218686.332 9034026.404 8469665.64 5643806.092 6347642.909 5840679.547 5428905.305 serine 75162158.92 79007926.19 82429768.99 75230778.37 103510439.4 118817481.9 113194754.6 105487761 succinate 10674150.94 10843572.99 10699288.22 9033596.024 5307124.332 5001079.5 4796129.301 4302733.665 succinyl-CoA 2303148.603 3801819.361 4171262.783 3986539.947 3491188.634 3523401.074 3379237.471 4014658.951 taurine 19934496.44 19518761.72 21036150.24 20329022.08 45896815.09 52064517.68 52097150.1 47004409.73 threonine 271549587.6 303606833.2 278197711.2 270475687.6 433939911.2 469843999.3 436202093.9 405425507.3 trans-urocanate 12475192.86 29512828.05 11408925.39 13189974.42 16511461.5 13126105.04 27970154.92 14907387.17 tryptophan 182172535 198910877 173513362.2 208448480.5 146266906.6 156058320.6 145500360 149280358.9 tyrosine 645378718 682729139.9 676474448.6 685725421.2 671969730.7 694066630.9 599134705 662233895.9 UDP-GlcNAc 143912309.2 154194459.1 165412695.3 159590995.4 103124768.5 100179694.7 91874951 103415823.3 UDP-hexose 123507732.6 122246139.6 123888906 120545833.8 87115893.69 63519720.52 66544126.93 85725456.87 UDP 12715640.27 13613538.78 12049437.94 13667090.58 9285389.209 10849743.88 10513689.46 8541087.179 UMP 2082092.344 1927133.453 1953234.332 1970349.345 1872456.047 1972835.239 2270354.912 1661091.143 Ureidosuccinic_acid 5910600.051 5828607.282 5931733.047 5775046.491 4983849.827 4529637.507 5178611.385 5115111.332 UTP 167450773.6 180309012.5 190587326.5 199315805.3 133692984.6 155068411.2 140311411.3 172946573.9 valine 413300322.6 407297465 367418208.1 476277958.8 391151441 405076016 353511873.1 370147826.6 MS_METABOLITE_DATA_END #METABOLITES METABOLITES_START metabolite_name Standardized name Formula Exact mass Super class Main class Sub class 2,3-Bisphosphoglycerate - 2-hydroxybutyric acid 2-Hydroxybutyric acid C4H8O3 104.0473 Fatty Acyls Fatty acids Hydroxy FA 2-hydroxyglutarate 2-Hydroxyglutaric acid C5H8O5 148.0372 Fatty Acyls Fatty acids Dicarboxylic acids 3-methyl-2-oxopentanoic acid 3-Methyl-2-oxovaleric acid C6H10O3 130.0630 Organic acids Keto acids Short-chain keto acids 3-phosphoglycerate 3-Phosphoglyceric acid C3H7O7P 185.9929 Organic acids Short-chain acids Short-chain acids 3-ureidopropionate Ureidopropionic acid C4H8N2O3 132.0535 Organic acids Carboximidic acids Ureas acetoacetate Acetoacetic acid C4H6O3 102.0317 Fatty Acyls Fatty acids Oxo FA Acetyl CoA Acetyl-CoA C23H38N7O17P3S 809.1258 Fatty Acyls Fatty esters Acyl CoAs acetylalanine N-Acetylalanine C5H9NO3 131.0582 Organic acids Amino acids and peptides Amino acids acetylaspartate N-Acetylaspartic acid C6H9NO5 175.0481 Organic acids Amino acids and peptides Amino acids acetylcarnitine CAR 2:0 C9H17NO4 203.1158 Fatty Acyls Fatty esters Acyl carnitines acetylglutamate N-Acetylglutamic acid C7H11NO5 189.0637 Organic acids Amino acids and peptides Amino acids acetylserine O-Acetylserine C5H9NO4 147.0532 Organic acids Amino acids and peptides Amino acids ADP Adenosine C10H13N5O4 267.0968 Nucleic acids Purines Purine ribonucleosides AICAR AICAR C9H14N4O5 258.0964 Nucleic acids Imidazoles Imidazole ribonucleosides and ribonucleotides alanine Alanine C3H7NO2 89.0477 Organic acids Amino acids and peptides Amino acids AMP AMP C10H14N5O7P 347.0631 Nucleic acids Purines Purine rNMP arginine Arginine C6H14N4O2 174.1117 Organic acids Amino acids and peptides Amino acids argininosuccinate unc - asparagine Asparagine C4H8N2O3 132.0535 Organic acids Amino acids and peptides Amino acids aspartate Aspartic acid C4H7NO4 133.0375 Organic acids Amino acids and peptides Amino acids asymmetric dimethylarginine Dimethylarginine C8H18N4O2 202.1430 Organic acids Amino acids and peptides Amino acids ATP ATP C10H16N5O13P3 506.9958 Nucleic acids Purines Purine rNTP beta-alanine beta-Alanine C3H7NO2 89.0477 Organic acids Amino acids and peptides Amino acids betaine Betaine C5H11NO2 117.0790 Organic acids Amino acids and peptides Amino acids biotin Biotin C10H16N2O3S 244.0882 Organoheterocyclic compounds Heterocyclic compounds Biotins Butyrylcarnitine CAR 4:0 C11H21NO4 231.1471 Fatty Acyls Fatty esters Acyl carnitines C5 Carnitines - cAMP 3',5' cyclic AMP C10H12N5O6P 329.0525 Nucleic acids Purines Purine rNMP carnitine Carnitine C7H15NO3 161.1052 Organic nitrogen compounds Carnitines Carnitines CDP CDP C9H15N3O11P2 403.0182 Nucleic acids Pyrimidines Pyrimidine rNDP choline Choline C5H14NO 104.1075 Organic nitrogen compounds Cholines Cholines cis-aconitate cis-Aconitic acid C6H6O6 174.0164 Organic acids TCA acids TCA acids citrate Citric acid C6H8O7 192.0270 Organic acids TCA acids TCA acids citrulline Citrulline C6H13N3O3 175.0957 Organic acids Amino acids and peptides Amino acids CMP CMP C9H14N3O8P 323.0519 Nucleic acids Pyrimidines Pyrimidine rNMP Coenzyme A Coenzyme A C21H36N7O16P3S 767.1152 Nucleic acids Purines Other purines creatine Creatine C4H9N3O2 131.0695 Organic acids Amino acids and peptides Amino acids creatinine Creatinine C4H7N3O 113.0589 Organoheterocyclic compounds Azoles Imidazolines CTP CTP C9H16N3O14P3 482.9845 Nucleic acids Pyrimidines Pyrimidine rNTP cystathionine Cystathionine C7H14N2O4S 222.0674 Organic acids Amino acids and peptides Amino acids cystine Cystine C6H12N2O4S2 240.0239 Organic acids Amino acids and peptides Amino acids cytidine Cytidine C9H13N3O5 243.0855 Nucleic acids Pyrimidines Pyrimidine ribonucleosides dATP dATP C10H16N5O12P3 491.0008 Nucleic acids Purines Purine dNTP dCTP dCTP C9H16N3O13P3 466.9896 Nucleic acids Pyrimidines Pyrimidine dNTP dihydroxyacetone phosphate Dihydroxyacetone phosphate C3H7O6P 169.9980 Organic acids Short-chain acids Short-chain acids dimethylglycine Dimethylglycine C4H9NO2 103.0633 Organic acids Amino acids and peptides Amino acids dTMP dTMP C10H15N2O8P 322.0566 Nucleic acids Pyrimidines Pyrimidine dNMP dUMP dUMP C9H13N2O8P 308.0410 Nucleic acids Pyrimidines Pyrimidine dNMP erythrose-4-phosphate Erythrose 4-phosphate C4H9O7P 200.0086 Carbohydrates Monosaccharides Monosaccharide phosphates F6P_G1P - FAD FAD C27H33N9O15P2 785.1571 Nucleic acids Flavins Flavin nucleotides folic acid Folic acid C19H19N7O6 441.1397 Organoheterocyclic compounds Pterins Folic acids formyl-methionine N-Formylmethionine C6H11NO3S 177.0460 Organic acids Amino acids and peptides Amino acids Fructose-1,6-bisphosphate Fructose 1,6-bisphosphate C6H14O12P2 339.9961 Carbohydrates Monosaccharides Monosaccharide phosphates fumarate Fumaric acid C4H4O4 116.0110 Organic acids TCA acids TCA acids GABA gamma-Aminobutyric acid C4H9NO2 103.0633 Fatty Acyls Fatty acids Amino FA GAR GAR C7H15N2O8P 286.0566 Nucleic acids Glycinamide ribonucleotides Glycinamide ribonucleotides GDP GDP C10H15N5O11P2 443.0243 Nucleic acids Purines Purine rNDP Glucose-6-phosphate Glucose 6-phosphate C6H13O9P 260.0297 Carbohydrates Monosaccharides Monosaccharide phosphates glucose Glucose C6H12O6 180.0634 Carbohydrates Monosaccharides Hexoses glutamate Glutamic acid C5H9NO4 147.0532 Organic acids Amino acids and peptides Amino acids glutamine Glutamine C5H10N2O3 146.0691 Organic acids Amino acids and peptides Amino acids glyceraldehyde-3-phosphate Glyceraldehyde 3-phosphate C3H7O6P 169.9980 Organic acids Short-chain acids Short-chain acids glycerate Glyceric acid C3H6O4 106.0266 Carbohydrates Monosaccharides Sugar acids Glycerol-3-Phosphate sn-Glycero-3-phosphate C3H9O6P 172.0137 Organic acids Organic phosphoric acids Organic phosphoric acids Glycerophosphocholine sn-Glycero-3-phosphocholine C8H20NO6P 257.1028 Organic acids Organic phosphoric acids Organic phosphoric acids glycine Glycine C2H5NO2 75.0320 Organic acids Amino acids and peptides Amino acids GMP GMP C10H14N5O8P 363.0580 Nucleic acids Purines Purine rNMP Glutathione Glutathione C10H17N3O6S 307.0838 Organic acids Amino acids and peptides Tripeptides Glutathione disulfide Oxidized glutathione C20H32N6O12S2 612.1520 Organic acids Amino acids and peptides Tripeptides GTP GTP C10H16N5O14P3 522.9907 Nucleic acids Purines Purine rNTP guanine Guanine C5H5N5O 151.0494 Nucleic acids Purines Hypoxanthines guanosine Guanosine C10H13N5O5 283.0917 Nucleic acids Purines Purine ribonucleosides histidine Histidine C6H9N3O2 155.0695 Organic acids Amino acids and peptides Amino acids hydroxyproline Hydroxyproline C5H9NO3 131.0582 Organic acids Amino acids and peptides Amino acids hypotaurine Hypotaurine C2H7NO2S 109.0198 Organic acids Sulfinic acids Sulfinic acids hypoxanthine Hypoxanthine C5H4N4O 136.0385 Nucleic acids Purines Hypoxanthines IMP IMP C10H13N4O8P 348.0471 Nucleic acids Purines Purine rNMP inosine Inosine C10H12N4O5 268.0808 Nucleic acids Purines Purine ribonucleosides isoleucine Isoleucine C6H13NO2 131.0946 Organic acids Amino acids and peptides Amino acids Itaconic Acid Itaconic acid C5H6O4 130.0266 Fatty Acyls Fatty acids Dicarboxylic acids kynurenine Kynurenine C10H12N2O3 208.0848 Organic acids Amino acids and peptides Amino acids lactic acid Lactic acid C3H6O3 90.0317 Organic acids Short-chain acids Short-chain acids leucine Leucine C6H13NO2 131.0946 Organic acids Amino acids and peptides Amino acids lysine Lysine C6H14N2O2 146.1055 Organic acids Amino acids and peptides Amino acids malate D-Malic acid C4H6O5 134.0215 Organic acids Hydroxy acids Beta hydroxy acids Methionine sulfoxide Methionine sulfoxide C5H11NO3S 165.0460 Organic acids Amino acids and peptides Amino acids methionine Methionine C5H11NO2S 149.0511 Organic acids Amino acids and peptides Amino acids methylthioadenosine Methylthioadenosine C11H15N5O3S 297.0896 Nucleic acids Purines Purine ribonucleosides myo-inositol Myo-inositol C6H12O6 180.0634 Organic oxygen compounds Alcohols and polyols Inositols N-acetylglutamate N-Acetylglutamic acid C7H11NO5 189.0637 Organic acids Amino acids and peptides Amino acids N-acetylglutamine N-Acetylglutamine C7H12N2O4 188.0797 Organic acids Amino acids and peptides Amino acids N-acetyllysine N(2)-Acetyllysine C8H16N2O3 188.1161 Organic acids Amino acids and peptides Amino acids NAD+ NAD+ C21H28N7O14P2 664.1170 Nucleic acids Nicotinamides Nicotinamide dinucleotides NADH NADH C21H29N7O14P2 665.1248 Nucleic acids Nicotinamides Nicotinamide dinucleotides NADP NADP+ C21H29N7O17P3 744.0833 Nucleic acids Nicotinamides Nicotinamide dinucleotides NADPH NADPH C21H30N7O17P3 745.0911 Nucleic acids Nicotinamides Nicotinamide dinucleotides niacinamide Niacinamide C6H6N2O 122.0480 Alkaloids Nicotinic acid alkaloids Pyridine alkaloids Nicotinic Acid Nicotinic acid C6H5NO2 123.0320 Alkaloids Nicotinic acid alkaloids Pyridine alkaloids O-phosphoethanolamine Phosphoethanolamine C2H8NO4P 141.0191 Organic acids Phosphate esters Phosphoethanolamines ornithine Ornithine C5H12N2O2 132.0899 Organic acids Amino acids and peptides Amino acids orotate Orotic acid C5H4N2O4 156.0171 Nucleic acids Pyrimidines Pyrimidine carboxylic acids palmitate Palmitic acid C16H32O2 256.2402 Fatty Acyls Fatty acids Saturated FA phenylalanine Phenylalanine C9H11NO2 165.0790 Organic acids Amino acids and peptides Amino acids phosphocholine Choline phosphate C5H14NO4P 183.0660 Organic nitrogen compounds Cholines Cholines phosphocreatine Phosphocreatine C4H10N3O5P 211.0358 Organic acids Amino acids and peptides Amino acids phosphoenolpyruvate Phosphoenolpyruvic acid C3H5O6P 167.9824 Organic acids Short-chain acids Short-chain acids phosphoethanolamine Phosphoethanolamine C2H8NO4P 141.0191 Organic acids Phosphate esters Phosphoethanolamines phosphoserine Phosphoserine C3H8NO6P 185.0089 Organic acids Amino acids and peptides Amino acids Pipecolate Pipecolic acid C6H11NO2 129.0790 Organic acids Amino acids and peptides Amino acids proline Proline C5H9NO2 115.0633 Organic acids Amino acids and peptides Amino acids propionylcarnitine CAR 3:0 C10H19NO4 217.1314 Fatty Acyls Fatty esters Acyl carnitines PRPP Phosphoribosyl pyrophosphate C5H13O14P3 389.9518 Carbohydrates Monosaccharides Monosaccharide phosphates Pyridoxal Pyridoxal C8H9NO3 167.0582 Alkaloids Nicotinic acid alkaloids Pyridine alkaloids Pyridoxine Pyridoxine C8H11NO3 169.0739 Alkaloids Nicotinic acid alkaloids Pyridine alkaloids pyruvate Pyruvic acid C3H4O3 88.0160 Organic acids Short-chain acids Short-chain acids Ribose 1-phosphate Ribose 1-phosphate C5H11O8P 230.0192 Carbohydrates Monosaccharides Monosaccharide phosphates Ribose_Ribulose 5-phosphate_3Me-Lys - S-Adenosyl-L-Homocysteine S-Adenosylhomocysteine C14H20N6O5S 384.1216 Nucleic acids Purines Other purines S-Adenosyl-L-methionine S-Adenosylmethionine C15H23N6O5S 399.1451 Nucleic acids Purines Other purines sarcosine Sarcosine C3H7NO2 89.0477 Organic acids Amino acids and peptides Amino acids Sedoheptulose-7-phosphate Sedoheptulose 7-phosphate C7H15O10P 290.0403 Carbohydrates Monosaccharides Monosaccharide phosphates serine Serine C3H7NO3 105.0426 Organic acids Amino acids and peptides Amino acids succinate Succinic acid C4H6O4 118.0266 Organic acids TCA acids TCA acids succinyl-CoA Succinyl-CoA C25H40N7O19P3S 867.1313 Fatty Acyls Fatty esters Acyl CoAs taurine Taurine C2H7NO3S 125.0147 Organic acids Sulfonic acids Sulfonic acids threonine Threonine C4H9NO3 119.0582 Organic acids Amino acids and peptides Amino acids trans-urocanate Urocanic acid C6H6N2O2 138.0429 Alkaloids Histidine alkaloids Imidazole alkaloids tryptophan Tryptophan C11H12N2O2 204.0899 Organic acids Amino acids and peptides Amino acids tyrosine Tyrosine C9H11NO3 181.0739 Organic acids Amino acids and peptides Amino acids UDP-GlcNAc UDP-N-acetylglucosamine C17H27N3O17P2 607.0816 Nucleic acids Pyrimidines Pyrimidine ribonucleosides UDP-hexose UDP-hexose C15H24N2O17P2 566.0550 Nucleic acids Pyrimidines Pyrimidine rNDP UDP UDP C9H14N2O12P2 404.0022 Nucleic acids Pyrimidines Pyrimidine rNDP UMP UMP C9H13N2O9P 324.0359 Nucleic acids Pyrimidines Pyrimidine rNMP Ureidosuccinic_acid N-Carbamoylaspartic acid C5H8N2O5 176.0433 Organic acids Amino acids and peptides Amino acids UTP UTP C9H15N2O15P3 483.9685 Nucleic acids Pyrimidines Pyrimidine rNTP valine Valine C5H11NO2 117.0790 Organic acids Amino acids and peptides Amino acids METABOLITES_END #END