#METABOLOMICS WORKBENCH amaynard_20231001_193832 DATATRACK_ID:4358 STUDY_ID:ST002898 ANALYSIS_ID:AN004757 PROJECT_ID:PR001773 VERSION 1 CREATED_ON October 1, 2023, 7:42 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, SHIN1 or AICAR ST:STUDY_TITLE treatment ST:STUDY_SUMMARY Culture of MEL cells in RPMI media containing 2000 nM folic acid for 48hr in the ST:STUDY_SUMMARY presence or absence of SHIN1 (1.25 uM) or AICAR (500 nM). In addition, K562 ST:STUDY_SUMMARY cells were cultured in 100 nM folic acid. These samples were followed up by ST:STUDY_SUMMARY metabolomics targeting 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_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_2000_nM_FA_Vehicle_1 Treatment:2000_nM_FA_Vehicle RAW_FILE_NAME=Copy of 20220615_AM_Inhibitor48hr_KM_AM769.raw SUBJECT_SAMPLE_FACTORS MEL MEL_2000_nM_FA_Vehicle_2 Treatment:2000_nM_FA_Vehicle RAW_FILE_NAME=Copy of 20220615_AM_Inhibitor48hr_KM_AM770.raw SUBJECT_SAMPLE_FACTORS MEL MEL_2000_nM_FA_Vehicle_3 Treatment:2000_nM_FA_Vehicle RAW_FILE_NAME=Copy of 20220615_AM_Inhibitor48hr_KM_AM771.raw SUBJECT_SAMPLE_FACTORS MEL MEL_100_nM_FA_Vehicle_1 Treatment:100_nM_FA_Vehicle RAW_FILE_NAME=Copy of 20220615_AM_Inhibitor48hr_KM_AM772.raw SUBJECT_SAMPLE_FACTORS MEL MEL_100_nM_FA_Vehicle_2 Treatment:100_nM_FA_Vehicle RAW_FILE_NAME=Copy of 20220615_AM_Inhibitor48hr_KM_AM773.raw SUBJECT_SAMPLE_FACTORS MEL MEL_100_nM_FA_Vehicle_3 Treatment:100_nM_FA_Vehicle RAW_FILE_NAME=Copy of 20220615_AM_Inhibitor48hr_KM_AM774.raw SUBJECT_SAMPLE_FACTORS MEL MEL_2000_nM_FA_AICAR_1 Treatment:2000_nM_FA_AICAR RAW_FILE_NAME=Copy of 20220615_AM_Inhibitor48hr_KM_AM775.raw SUBJECT_SAMPLE_FACTORS MEL MEL_2000_nM_FA_AICAR_2 Treatment:2000_nM_FA_AICAR RAW_FILE_NAME=Copy of 20220615_AM_Inhibitor48hr_KM_AM776.raw SUBJECT_SAMPLE_FACTORS MEL MEL_2000_nM_FA_AICAR_3 Treatment:2000_nM_FA_AICAR RAW_FILE_NAME=Copy of 20220615_AM_Inhibitor48hr_KM_AM777.raw SUBJECT_SAMPLE_FACTORS MEL MEL_2000_nM_FA_SHIN1_1 Treatment:2000_nM_FA_SHIN1 RAW_FILE_NAME=Copy of 20220615_AM_Inhibitor48hr_KM_AM778.raw SUBJECT_SAMPLE_FACTORS MEL MEL_2000_nM_FA_SHIN1_2 Treatment:2000_nM_FA_SHIN1 RAW_FILE_NAME=Copy of 20220615_AM_Inhibitor48hr_KM_AM779.raw SUBJECT_SAMPLE_FACTORS MEL MEL_2000_nM_FA_SHIN1_3 Treatment:2000_nM_FA_SHIN1 RAW_FILE_NAME=Copy of 20220615_AM_Inhibitor48hr_KM_AM780.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 48hr in 2000 nM folic acid + vehicle, 100 nM folic TR:TREATMENT_SUMMARY acid + vehicle, 2000 nM folic acid + AICAR (500 nM), and 2000 nM folic acid + TR:TREATMENT_SUMMARY SHIN1 (1.25 uM). #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_2000_nM_FA_Vehicle_1 MEL_2000_nM_FA_Vehicle_2 MEL_2000_nM_FA_Vehicle_3 MEL_100_nM_FA_Vehicle_1 MEL_100_nM_FA_Vehicle_2 MEL_100_nM_FA_Vehicle_3 MEL_2000_nM_FA_AICAR_1 MEL_2000_nM_FA_AICAR_2 MEL_2000_nM_FA_AICAR_3 MEL_2000_nM_FA_SHIN1_1 MEL_2000_nM_FA_SHIN1_2 MEL_2000_nM_FA_SHIN1_3 Factors Treatment:2000_nM_FA_Vehicle Treatment:2000_nM_FA_Vehicle Treatment:2000_nM_FA_Vehicle Treatment:100_nM_FA_Vehicle Treatment:100_nM_FA_Vehicle Treatment:100_nM_FA_Vehicle Treatment:2000_nM_FA_AICAR Treatment:2000_nM_FA_AICAR Treatment:2000_nM_FA_AICAR Treatment:2000_nM_FA_SHIN1 Treatment:2000_nM_FA_SHIN1 Treatment:2000_nM_FA_SHIN1 hydroxyglutarate 433784601 451689177 421742493 253745318 245379857 254784877 158925166 160946011 213069262 292849862 329273861 330521160 oxoadipate 19728930 17860090 18286346 13429085 12772795 16087048 7765377 7735908 8702849 12315454 13043649 17298538 2,3-BPG 7109549 8229339 6591420 12221804 11735150 9891335 3567241 3822692 3545120 6932837 6723614 5919626 3-methyl-2-oxopentanoic acid 4668770 3595317 5448795 5682402 4832158 5077668 3694871 4637825 6728867 3055011 4205961 5077581 3-phosphoglycerate 54639741 59434700 53627112 63826161 65060536 65065295 27134655 28595957 23743252 116219500 113918556 111581866 6-phosphogluconate 2868279 2907031 2433361 3190336 2997353 3239548 1033242 1137060 1033639 2300546 2176337 2372184 Acetyl-CoA 45521731 49534764 54204620 54487048 56506986 50321714 38604804 39320260 34444739 37860993 35533534 29924370 acetylcarnitine 76634430 63070622 66331908 57137549 49135724 52496022 51593099 56297432 35243039 43090720 48121457 43759900 ADP-ribose 4415862.6 4476604.6 4210401.3 4048103.1 3418767.5 3989404.9 1211875.7 1078415.3 1057961.9 2046479.6 2011865.8 2164063.1 ADP 24282105 26863003 24905014 19771898 20339414 18239040 13379468 14178505 11796112 10433473 10516854 12262393 AICAR 64778365 85396438 56907274 98979709 105045946 114152172 1214129615 1175724105 1734642530 126571307 126376707 122692945 alanine 921241215 862135379 1040266803 1002530036 910248861 953917994 691840416 644910791 768771179 1111887383 1100324521 1029815485 Alpha-Ketoglutarate 17500042 14836415 21639276 14572375 11514301 12276300 6788005 11983240 8559132 23508939 20607701 36651040 AMP 27648516 25964670 25769233 23104026 19862987 19007255 18925014 21360816 19000238 14873453 14012725 14413464 arginine 8104168577 8639063819 9677014563 7738535913 7605678402 7851956594 4082455505 3939039778 5015976314 5841919643 5759154842 5347790894 argininosuccinate 60978352 57809600 63846062 56849597 53315254 54301673 9576201 8622284 10750906 10690499 10547077 12289387 ascorbate 3838264755 3970573559 3748073516 2988506421 3120322222 3329846863 2202543592 2251280851 2502047245 3099914997 3116278676 3393322233 asparagine 1525108071 1449308232 1597695037 1382614138 1353641223 1415865548 753271471 676791288 837190117 1081331890 1091321583 1030196891 aspartate 300966009 307660856 328822191 407706480 358366002 385998578 956954865 950347426 1099707273 894857156 866326304 846950827 ATP 479229956 437411306 456426468 452033116 691001408 425977907 375409367 592199747 633822526 224866628 213408699 204345368 beta-alanine 153287255 143845811 149947334 149769084 148304879 148914628 90214755 87349828 105383460 185356410 186237554 168420014 betaine 607039864 60307315 68298605 42776601 66489594 42836768 71082169 31406666 44453718 90750670 289650056 61928795 Butyrylcarnitine 33867275 31839172 39546396 39372096 38991889 43564565 20160297 20390294 12052613 23981185 24229827 27132853 C5-Carnitines 1543103.8 1615103.2 1089890.8 1306908.3 574665.3 808330.1 1487760 1114685 NA NA 765509.4 NA carnitine 183475964 175956937 194316584 213267310 210703828 206180489 185844514 165383460 172464450 181634792 193832461 178669370 CDP-choline 1750669.7 1648998.7 1866733.7 2406542.6 2478810.1 2495428.8 6431894.1 5493470.8 8509753.3 6225540.6 5632639 5654658.8 CDP 6683494.5 6366227.2 5246340.7 6307962.4 7402656.2 7222726.6 1389579.5 1844074.1 1513200.1 3694173.9 3683508.9 3395185.8 choline 208837791 206877252 220161518 165272812 150792842 159519979 184950899 170812448 172987573 119467246 112674573 cis-aconitate 104138320 101848759 108032658 93802220 103000288 103122836 42987510 41583727 40125582 76582901 76204662 75310614 citrate 2688339544 2912310199 2980040816 2757894793 2711239144 2881578178 1318870756 1400193393 1463687047 2288409935 2287004399 2239928967 CMP 1509616.2 1579915.5 1331672.1 1781077.7 1398690.6 1785485.9 1753672 756401.3 1719765.8 853008.8 1552190.2 1404335.6 creatine 90473983 87228833 85398132 64534646 59288520 54612417 50431284 49522937 64089620 119539090 155595732 144261736 CTP 109050617 105011952 96234198 144597108 213780345 129276423 24728163 36534562 35312690 74681445 71734750 69007699 cystathionine 10362157 7624898 8492161 22089936 18018425 21050806 2498131 2512359 2582468 11705840 11065501 13015407 cysteic acid 2701540.1 2937367.1 3022988.1 3676810.1 3005064.3 4129983.9 947793.8 792696.8 800929 2668911.8 2503645.2 819701.8 cystine 8943602 10988471 9712920 11564622 11769644 8690834 27996047 31479709 31293378 22160534 34126490 26806141 dATP 3386695.7 3312247.2 3283405.1 3461688.2 4792423.5 3353234.2 4107085.2 5443951 5514368.6 1526451.1 1409940.8 1123822.9 dihydroxyacetone phosphate 32172151 23148079 17338946 10774170 14456745 10073192 7059811 8503051 14507347 13707675 17970464 26115728 dUMP 3496864 3013329 4181195 F6P_G1P 31138573 31894766 34822039 36013120 36306268 35776251 21894729 21331386 22681200 19963040 18501857 18514170 formyl-methionine 1537918.5 1886002.5 2250966.2 4312205.5 4274883.2 5174584 968133.9 1127121.7 1480074.2 875402.9 144764.4 Fructose-1,6-bisphosphate 82757085 78221781 76438152 89844836 92653950 83253185 28637231 29505512 27667708 50953690 48036042 46212950 fumarate 98704500 94816452 98431351 110406130 95317443 108700615 52815399 53059442 43820145 43616582 39552558 42016948 gamma-aminobutyric acid 92788015 99505365 87528540 59685336 68458070 68277577 43226537 47229363 50576327 117900064 139022645 119737339 GAR 15062526 18298429 13884206 37759734 39128277 43241191 57884456 55354697 54168470 GDP 12619489 11211677 11762828 11615460 12126579 11607137 8371900 9942271 9092328 7990031 7414410 6962489 Glucose-6-phosphate 26158091 26304096 27221763 35412462 33794577 36110465 22212964 18915448 24384545 23935484 23042878 24068777 glucose NA 10804424 NA 13690746 11751956 10802351 14648565 13253560 16279078 20807712 19921135 22099821 glutamate 3783001027 3971871229 4128812730 4520463159 4144374294 4523398654 2719407353 2627397999 2776062762 6215014276 5979286459 6171346659 glutamine 6630502611 6857409111 7401683257 4912785782 5093849766 5586073939 6480637584 5972814699 7659291740 7670713146 7940447580 7848812927 glycerate 12200255525 12637714797 12579606628 9043733211 8644916452 9416530370 5515282223 5737684748 6204472832 8766173356 9121347969 9640880812 Glycerol-3-Phosphate 1501402 1817619.7 NA 1424384.7 NA 1822381.2 NA 682380 898109.7 NA 1029825.4 NA Glycerophosphocholine 11365223658 11485381943 12139328198 9630608895 8968850363 9112761137 9776641140 9402299866 10060661312 5745877916 4862475637 5003908531 glycine 94259116 89784730 92760836 65611688 71905310 60718627 128742504 121775649 147231158 63276459 64166633 59199855 GSSG 1075801952 1104990737 1199197071 988424578 916743876 925708746 921721108 991204722 1168246895 1363991862 1421118739 1416499879 GTP 71342001 70685968 66845434 82925745 132641298 71971545 50654243 77910085 83124709 41621524 40375226 37803182 histidine 569170448 568086332 572483069 538412323 476036990 561722910 353820203 360386813 398931025 966548608 1094229752 1036703092 hydroxyproline gammaALA 386542380 380910797 437129396 468447005 396745480 455559175 236560272 208158095 230271828 319284352 292334973 309051520 hypoxanthine 17509782 14791885.8 20750535.4 16197415.3 12270541.9 13766040.8 1548771571 1215023658 976948103.2 5888640.7 7917967.7 11939572.1 IMP 9438562 10993808 9413189 9147715 9682311 9299644 11264800 11160551 13125748 11151338 10826777 10069777 inosine 5074501.9 3604899 4254136 3420790.7 3020999.8 2405728.8 30166409.5 29140826.5 21495327.9 1274044.3 1473651.7 1269401.4 isoleucine 1431468253 1331651295 1499163415 1308249437 1199532898 1352244808 920888087 903105148 742079327 1534661280 1586802066 1500629433 Itaconic Acid 369016726 401393459 391199588 283270159 301121641 298922998 174341374 190278051 170466921 289816476 288286928 275598890 kynurenine 8463367 7694489 8141518 8910096 6042262 8028483 3051096 2156133 1195063 6589776 6954212 3690312 lactic acid 2685457163 2870013372 2873283169 1988235928 1531006291 1982992062 1224077632 1291289384 878958949 1972613619 2035620235 2037435773 leucine 962202768 863659236 951177493 933235504 762139979 903112810 807531631 749388170 607361256 1396834828 1439122400 1458359483 lysine 44839443 26031009 31650476 25173834 30731638 28812795 108394959 108030397 151378792 109779079 127143654 116570366 malate 1285860073 1334545670 1370166819 1403635351 1238402018 1415480018 712603534 723589217 630827742 624931512 642641702 614790432 Methionine sulfoxide 1378584668 1345036432 1423779994 1101510125 1059563627 1142580445 1289309251 1200101182 1338626731 2142813276 2126475068 2046233866 methionine 75561998 77827559 93186547 83087650 89719344 94786471 161420951 153284574 161192070 220485054 209677538 329665609 methylthioadenosine 34446358 36654214 37999587 35977684 26585075 34154193 19518927 19347755 10756232 30051381 36867208 48799392 N-acetylaspartate NA NA NA NA NA NA NA NA 424498.8 2237743.7 964615.6 426507.5 N-acetylglutamate 39725602 47543035 49526481 37112087 41286298 38987179 47506184 50399974 45308800 23846610 28762115 21762537 N-acetylphenylalanine 688383.6 715002.5 266847.1 620095.8 498735.1 607265 556184.1 616879.2 368159 349990 558638.3 607469.5 NAD 669642312 645444904 688853611 630781449 683195634 674661151 347580158 359920159 378397510 691752834 636030350 578063761 NADP 24793637 21524567 23005598 25382566 27953417 23694693 21071786 20936247 24047568 27530266 25619368 23586338 niacinamide 236015902 196380670 306312408 300739977 297522865 420252186 128381930 120884608 81245561 283005692 258050827 333040062 O-phosphoethanolamine 447627.8 508881.6 524911.1 467481.7 506608.5 433284.1 10151399.8 8504254.7 15451417.6 1227109.3 1275888.4 1411501.7 ornithine 30318961 28571623 34560757 17412023 18245020 18862963 13077550 11530320 22434959 32982911 32353277 38003347 orotate 5684589 5460482 5383121 3301694 3495214 3392092 448384523 492865396 367646793 3604302 3830401 4171857 palmitate 204163308 288499105 187125416 148138266 118935954 141853728 74940255 79072121 60975023 143050059 138349244 121927580 Pantothenate 425448779 440151568 421426046 570896656 441473421 525343144 547539772 559044872 361136739 1311078541 1294806203 1327763014 phenylalanine 25377027 17489876 16916922 12549954 10818041 12211765 70871460 69451959 47448494 97693729 105006635 102260522 phosphocholine 25718122748 27785970560 28555365616 26589647495 27835411726 28253372369 29966577934 29427281780 32099418459 25290693866 24319718321 23014229860 phosphocreatine NA NA NA NA NA NA NA NA NA NA NA NA phosphoenolpyruvate 18423741 21897591 19744495 23619050 23276002 22279065 13304063 13130563 14083934 57239756 56445212 55229044 proline 16987297557 17385441176 18393558384 17702274233 15479269882 16767704248 7309096018 6500485107 7182317888 12536923752 12061809556 11887163589 propionylcarnitine 36694736 32905538 34688128 20945075 17686801 15366200 17569184 15816075 13856744 15434955 18672931 18513880 PRPP 10685838 13796998 8007348 23356547 31192834 24871563 2041290 1364103 2537106 64877205 65728708 57230729 Pyridoxine 118586524 118830266 133192814 158135633 117104009 150281239 136312167 131714875 103595517 201977266 207374446 239160856 Ribose-1-phosphate 5747168 5992225 5029163 14858057 15051675 15532272 1199320 1113817 1026900 20482982 20053684 19244522 Ribose_Ribulose-5-phosphate-3Me-Lys 8328016 9138990 6289711 12204786 13284564 13519052 2954684 3825168 5918052 24809060 25097755 26638622 S-Adenosyl-L-Homocysteine 14065466 15463127 15038685 14523546 15273189 14794763 32452559 33891848 37583584 14322625 13917134 14976000 S-Adenosyl-L-methionine 1107914047 1135172737 1140243135 1192403647 1118339097 1138811868 532858226 534210691 590657692 952509205 1005825440 984277889 Sedoheptulose-7-phosphate 39991825 42161196 42541025 39183635 37084380 40279962 9732141 9618671 9828346 4270944 3968168 3474103 serine 59136494 52152173 59674411 110211629 108020072 105382378 121529511 109695627 139564975 717704068 659820208 650972735 succinate 50021775 55335615 54738161 57602818 51978221 57778086 26723917 30292694 27455056 34322441 38500397 40150613 succinyl-CoA 6456490 7275133 7394593 5608179 7375022 4999573 6417176 7247578 6686987 4350075 4224605 3165027 taurine 7327411 8157685 8387252 8314556 8650234 8160826 8538994 8946146 9456993 9757663 9674957 8080893 threonine 455247263 448792415 445995763 396963167 378840031 376006337 475960412 479442039 561928721 775950417 775905479 775238018 tryptophan 24096881 23464442 24039223 15482276 16122346 17550046 77546188 76446188 80997896 137030074 142943389 170962998 tyrosine 440059532 467753368 503029920 473115849 449602534 472763413 393725182 389782298 373331352 682708263 677355336 704425335 UDP-GlcNAc 270818144 269223200 258502281 342449984 348622983 319491618 83805159 93597459 77303183 206003411 193585686 184739617 UDP 13928159 13831173 12959114 16353136 20926500 17701249 2130304 3083245 3058418 14956447 14869558 15528254 UMP 3276212 3887189.5 3671667.9 4205901.8 4230822.6 4172734.8 861942.1 1474460.9 917125 3341970.5 3287839.8 3401993.8 Ureidosuccinic acid 5961402 6728951 6347570 4188317 5108181 4906220 119308232 137229630 137721597 3853609 4861685 4043497 UTP 298148798 298523456 260067294 409097422 600371923 389464650 63817413 97790165 83658332 318552142 316333842 284994774 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 oxoadipate Oxoadipic acid C6H8O5 160.0372 Dicarboxylic acids 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-ribose ADP-ribose C15H23N5O14P2 559.0717 Purine rNDP ADP ADP C10H15N5O10P2 427.0294 Purine rNDP AICAR AICAR C9H14N4O5 258.0964 Imidazole ribonucleosides and ribonucleotides alanine Alanine C3H7NO2 89.0477 Amino acids Alpha-Ketoglutarate Oxoglutaric acid C5H6O5 146.0215 TCA 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 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 CDP CDP C9H15N3O11P2 403.0182 Pyrimidine rNDP choline Choline C5H14NO 104.1075 Cholines cis-aconitate cis-Aconitic acid C6H6O6 174.0164 TCA acids citrate Citric acid C6H8O7 192.0270 TCA acids CMP CMP C9H14N3O8P 323.0519 Pyrimidine rNMP creatine Creatine C4H9N3O2 131.0695 Amino acids CTP CTP C9H16N3O14P3 482.9845 Pyrimidine rNTP cystathionine Cystathionine C7H14N2O4S 222.0674 Amino acids cysteic acid Cysteic acid C3H7NO5S 169.0045 Amino acids cystine Cystine C6H12N2O4S2 240.0239 Amino acids dATP dATP C10H16N5O12P3 491.0008 Purine dNTP dihydroxyacetone phosphate Dihydroxyacetone phosphate C3H7O6P 169.9980 Short-chain acids dUMP dUMP C9H13N2O8P 308.0410 Pyrimidine dNMP F6P_G1P - - - - 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 gamma-aminobutyric acid gamma-Aminobutyric acid C4H9NO2 103.0633 Amino FA 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 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 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 GSSG Oxidized glutathione C20H32N6O12S2 612.1520 Tripeptides GTP GTP C10H16N5O14P3 522.9907 Purine rNTP histidine Histidine C6H9N3O2 155.0695 Amino acids hydroxyproline gammaALA - - - - hypoxanthine Hypoxanthine C5H4N4O 136.0385 Hypoxanthines IMP IMP C10H13N4O8P 348.0471 Purine rNMP inosine Inosine C10H12N4O5 268.0808 Purine ribonucleosides isoleucine Isoleucine C6H13NO2 131.0946 Amino acids Itaconic Acid Itaconic acid C5H6O4 130.0266 Dicarboxylic acids kynurenine Kynurenine C10H12N2O3 208.0848 Amino acids lactic acid Lactic acid C3H6O3 90.0317 Short-chain 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 N-acetylaspartate N-Acetylaspartic acid C6H9NO5 175.0481 Amino acids N-acetylglutamate N-Acetylglutamic acid C7H11NO5 189.0637 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 ornithine Ornithine C5H12N2O2 132.0899 Amino acids orotate Orotic acid C5H4N2O4 156.0171 Pyrimidine carboxylic acids palmitate Palmitic acid C16H32O2 256.2402 Saturated FA 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 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 S-Adenosyl-L-methionine S-Adenosylmethionine C15H23N6O5S 399.1451 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 succinyl-CoA Succinyl-CoA C25H40N7O19P3S 867.1313 Acyl CoAs 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 METABOLITES_END #END