#METABOLOMICS WORKBENCH amaynard_20231001_192427 DATATRACK_ID:4357 STUDY_ID:ST002897 ANALYSIS_ID:AN004756 PROJECT_ID:PR001773 VERSION 1 CREATED_ON October 1, 2023, 7:31 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, SHIN1 or AICAR ST:STUDY_TITLE treatment ST:STUDY_SUMMARY Culture of K562 cells in RPMI media containing 2000 nM folic acid for 48hr in ST:STUDY_SUMMARY the 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 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_2000_nM_FA_Vehicle_1 Treatment:2000_nM_FA_Vehicle RAW_FILE_NAME=Copy of 20220615_AM_Inhibitor48hr_KM_AM754.raw SUBJECT_SAMPLE_FACTORS K562 K562_2000_nM_FA_Vehicle_2 Treatment:2000_nM_FA_Vehicle RAW_FILE_NAME=Copy of 20220615_AM_Inhibitor48hr_KM_AM755.raw SUBJECT_SAMPLE_FACTORS K562 K562_2000_nM_FA_Vehicle_3 Treatment:2000_nM_FA_Vehicle RAW_FILE_NAME=Copy of 20220615_AM_Inhibitor48hr_KM_AM756.raw SUBJECT_SAMPLE_FACTORS K562 K562_100_nM_FA_Vehicle_1 Treatment:100_nM_FA_Vehicle RAW_FILE_NAME=Copy of 20220615_AM_Inhibitor48hr_KM_AM757.raw SUBJECT_SAMPLE_FACTORS K562 K562_100_nM_FA_Vehicle_2 Treatment:100_nM_FA_Vehicle RAW_FILE_NAME=Copy of 20220615_AM_Inhibitor48hr_KM_AM758.raw SUBJECT_SAMPLE_FACTORS K562 K562_100_nM_FA_Vehicle_3 Treatment:100_nM_FA_Vehicle RAW_FILE_NAME=Copy of 20220615_AM_Inhibitor48hr_KM_AM759.raw SUBJECT_SAMPLE_FACTORS K562 K562_2000_nM_FA_AICAR_1 Treatment:2000_nM_FA_AICAR RAW_FILE_NAME=Copy of 20220615_AM_Inhibitor48hr_KM_AM760.raw SUBJECT_SAMPLE_FACTORS K562 K562_2000_nM_FA_AICAR_2 Treatment:2000_nM_FA_AICAR RAW_FILE_NAME=Copy of 20220615_AM_Inhibitor48hr_KM_AM761.raw SUBJECT_SAMPLE_FACTORS K562 K562_2000_nM_FA_AICAR_3 Treatment:2000_nM_FA_AICAR RAW_FILE_NAME=Copy of 20220615_AM_Inhibitor48hr_KM_AM762.raw SUBJECT_SAMPLE_FACTORS K562 K562_2000_nM_FA_SHIN1_1 Treatment:2000_nM_FA_SHIN1 RAW_FILE_NAME=Copy of 20220615_AM_Inhibitor48hr_KM_AM763.raw SUBJECT_SAMPLE_FACTORS K562 K562_2000_nM_FA_SHIN1_2 Treatment:2000_nM_FA_SHIN1 RAW_FILE_NAME=Copy of 20220615_AM_Inhibitor48hr_KM_AM764.raw SUBJECT_SAMPLE_FACTORS K562 K562_2000_nM_FA_SHIN1_3 Treatment:2000_nM_FA_SHIN1 RAW_FILE_NAME=Copy of 20220615_AM_Inhibitor48hr_KM_AM765.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 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 K562_2000_nM_FA_Vehicle_1 K562_2000_nM_FA_Vehicle_2 K562_2000_nM_FA_Vehicle_3 K562_100_nM_FA_Vehicle_1 K562_100_nM_FA_Vehicle_2 K562_100_nM_FA_Vehicle_3 K562_2000_nM_FA_AICAR_1 K562_2000_nM_FA_AICAR_2 K562_2000_nM_FA_AICAR_3 K562_2000_nM_FA_SHIN1_1 K562_2000_nM_FA_SHIN1_2 K562_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 204093221 203452237 160809079 182400524 155162676 156960067 172014845 157195491 141682354 156723640 166631475 202912181 oxoadipate 10553013 10255148 9813508 9317378 10052149 8844241 9324020 10243134 8451021 9107745 9539326 9882995 2,3-BPG 29152767 29585951 31680827 24026862 26227021 26079764 6791978 7831123 9684527 13304549 13661574 12445476 3-methyl-2-oxopentanoic acid 31602628 27381660 30066670 15449395 14291088 20783249 17734996 13704302 12836375 17001501 18421486 12212898 3-phosphoglycerate 73304539 76424185 72849330 61612558 58439618 59807612 18281624 21231980 20450704 37509827 38696912 40842701 6-phosphogluconate 16524036 17139133 15780449 9541720 10041618 10225432 9241042 10523607 9964968 4277237 4225811 4391027 Acetyl-CoA 30072031 33434871 33356978 28253205 30910408 28347141 26594432 29254822 30355877 15613235 15945217 15905455 acetylcarnitine 171043187 165396468 167113132 207208967 197845595 206266618 385674158 394380543 313426143 106511770 107596846 97865221 ADP-ribose 1841592.8 1788283.6 1091298.2 1798768.7 1430110.7 1108941.1 798694.6 1020872.1 758440.5 856106.6 862649.2 1371893.8 ADP 14983882 15146965 11960692 10819980 9985158 13068352 12847711 13432204 10670412 4745377 5787232 6221713 AICAR 78918826 79071803 72235967 603978995 571688923 574682501 3296138238 3405575084 3579021336 434244621 438331940 457137436 alanine 18336802 19722668 24185295 27289425 21682080 20128764 26595037 22611269 27801316 88640548 76604175 72521200 Alpha-Ketoglutarate 11732364 12069659 10123774 14801800 11163017 11540949 10461062 8824013 7941610 10517486 11753834 14159326 AMP 18097314 18196334 17665119 12890441 12972808 12157334 14199654 13859665 14569374 10014704 9698938 9732679 arginine 4838503429 4873354162 4894630256 5935890585 5589448536 5668526159 6214944080 5907987436 6404286224 5661772438 5441035190 5604200605 argininosuccinate 45090362 44146933 43476358 36956349 32213183 33041931 11642422 12129495 13852709 26452275 24806721 29132556 ascorbate 3135082058 3187974220 2935741044 2969155671 3239786421 2864931491 2875632951 3104235050 3172346529 2807813448 2814977305 2819519132 asparagine 785639618 698278692 764498653 854629687 791867244 796518697 832865157 831479418 914185618 603408528 567860582 604040590 aspartate 287702346 270371842 282559926 419555386 391842354 396257161 552397031 591420733 583938725 634699734 593807425 649138150 ATP 382934995 389730573 553045369 259925080 383317904 376331891 312240180 352807845 514115829 124395008 140309082 122762909 beta-alanine 288501553 282141400 284712488 247359678 253442295 237290173 235641568 246242387 266112179 281717437 305183885 299992191 betaine 397419833 363398192 382589585 590571716 644475173 646741129 860840329 856884738 717193482 1110428444 1252911828 1098571321 Butyrylcarnitine 23134499 22773497 23392626 22083694 20125643 23425406 16313515 17723767 14511151 9793656 10151855 10499987 C5-Carnitines 149226558.8 146926131.7 141960605.7 154058708.3 148422750.4 151961689.8 129525846 132923148.2 122367786.6 185453368.7 188183015.5 179515269.1 carnitine 280520324 277373905 284786575 247313293 251347211 241775382 280480841 256049615 285581798 246510448 238590853 211573135 CDP-choline 813053.4 865894.4 904226.7 730040.8 396172.6 879505.4 1300956.9 1192431.4 1287823.6 2915686.2 2556202.6 3019413.6 CDP 3582123 3963316.4 3646365.9 3825255 3600715.9 3506173.5 445426 872901.1 1516537.6 2467033.3 2518749.1 2487264.1 choline 172320739 164974130 146301646 87185198 1809974 3694374 2255565 cis-aconitate 59008147 54382675 52727428 53546245 56478764 62440971 50611047 52400987 60495133 86916933 76450494 74832406 citrate 1753743459 1867291914 1587588246 1937225205 1792427642 1757141286 1934787545 1948188949 1975649387 2289661103 2100490167 2330642087 CMP 478923.8 1658873.9 509936.4 418859.7 1562466.6 1367678.2 1472759.1 217617.9 791211.9 3093621.7 2924823.7 3042800.9 creatine 240751880 250988712 234569051 308522296 334058093 313761339 472748506 535800798 529322986 592399062 644739573 619247907 CTP 75422994 76723679 109709939 71976578 105085774 103565703 16232153 18692156 25562068 40423819 46694279 39484002 cystathionine 311879337 306321006 303423676 523716277 431167542 452094772 45609032 49097374 44335463 105338382 91249392 116266444 cysteic acid 788094.2 960360.9 765597.6 1234656.7 1005932.1 1415769.8 797675.2 777390.4 851837.2 1309442.5 1668319.6 1687444.2 cystine 9481195 5365036 7486222 7449609 6774616 12594427 24365890 17348923 22216202 33037468 36619685 18687224 dATP 4101773.3 4544138.5 5709648.8 2885590 3872710.4 3745369.1 3055515 2988167.2 3884971.9 853900.8 971442.3 811465 dihydroxyacetone phosphate 21697915 28887544 25401908 11526264 13887475 11316535 8759195 8153426 8842776 4374678 5036103 3928225 dUMP 6013023 5740849 5092530 35248013 34121317 29346411 F6P_G1P 41585387 43002448 42010620 40134085 36319280 38015818 87488361 88957358 86234772 23741821 23452720 24442295 formyl-methionine 10597642.4 10369146.3 10848217.5 8218931.1 7449130.3 9241902.6 6586773.9 6371377.1 5252367.2 4278511.8 5200028.7 4446545.5 Fructose-1,6-bisphosphate 75241468 78443923 81495057 53513527 52114957 52429171 25627482 28252588 29438373 51601717 56802075 53468846 fumarate 136981334 140139106 123428962 84756905 85086375 76197718 39251406 37229832 37704277 53032482 44314273 51967151 gamma-aminobutyric acid 45713152 42345245 45676451 46598954 40624953 44274652 44886519 43829785 54093730 145703323 145369624 154598044 GAR 6628166 6726647 6102465 68353407 66103000 62877272 83152053 80872727 90743570 GDP 10470891 10698282 10365456 6642224 6605850 6113290 6030797 5998291 7203064 4179195 4431097 4195905 Glucose-6-phosphate 47159277 50453166 50870764 52748718 50173317 50614957 129135457 122667240 123834517 34174871 32126120 32864759 glucose 8763128 NA 10260819 16831758 17738997 14748125 38278111 36564602 33088432 24732904 27648929 23867482 glutamate 2575294825 2444118867 2400695333 2815605412 2675603814 2647611897 3049786114 3095858220 3142372922 2722371465 2508908598 2745477219 glutamine 11246135868 10552931521 11144415386 12445751942 12719510377 12258232216 14644433821 14906859412 16065538540 11735753482 11840011996 12337438232 glycerate 7145059205 7021510016 6440322958 6851677724 7446209257 6442901496 6422624684 6598884055 6766347801 6366212005 6479800381 6724683754 Glycerol-3-Phosphate 10458841.1 9892987.4 10311896.3 6961542.5 7433703.2 8186639.6 4650481.4 4691949.5 5024110.3 2941862.9 2872830.2 3186130.7 Glycerophosphocholine 8045013543 7633849153 7777636466 3588289215 3627426255 3541708340 549393924 577787767 614686818 913050406 887956810 858104400 glycine 90761296 77758053 90218313 54505974 59705047 59664918 122814669 130479881 146007554 47015016 41519485 46260351 GSSG 1355889153 1318437825 1176582167 1225565539 1134200447 861109920 1496697854 1561288357 1463963822 1009924557 898040028 999712456 GTP 68334225 66602772 96106058 41857717 66509201 65503923 50178414 55251221 84433491 23329346 25178424 22879379 histidine 1157569076 1073221793 1011064064 1977174045 1700393108 1924141216 2764358970 2854177731 2863291118 2279261513 2237911932 2401190944 hydroxyproline gammaALA 213448571 196261154 202227425 255109619 232863512 239743710 264654607 255632809 271756862 174342819 157341489 177930181 hypoxanthine 3685244.3 2293265.4 2175421.7 2085125.3 2163665.9 1433306.5 115220748.1 118200399.9 75343618.1 2048021.8 2265818 1869072 IMP 5597177 5809694 5772705 5955274 5949483 6177019 8283894 8824455 9344834 2900822 3058968 2862026 inosine 193767.4 227839.4 265433.9 152481.4 208688.7 188986.1 1036747.8 1149815.8 923739.9 210666.2 225932.5 182208.9 isoleucine 2399811867 2138481655 2407869081 2552633675 2334426298 2517178513 2713969621 2895228543 2306367209 2508795426 2362862358 2159619029 Itaconic Acid 227116225 221134880 212748784 199202429 240886662 214268881 207378522 217095924 175997636 205234882 211405746 211651898 kynurenine 10612758 9882625 11852734 13428560 10426290 14092679 7302425 7234534 5932321 9651688 8064649 9559672 lactic acid 1518505136 1542790829 1405604440 1529124248 1610741361 1366636166 1428041207 1467171453 992114100 1425534991 1494924461 1572516556 leucine 2147510364 1888777113 2209182451 2415604918 2306146247 2515759652 2647884979 2481203806 2140330254 2461229213 2307114675 2358340455 lysine 63689920 66293051 71392199 101261972 91431198 103233987 214130986 184486783 208964532 184060874 165302019 183811663 malate 1713197903 1703037136 1594367716 1104932864 1139835025 987751010 552502668 543690372 566562146 725264792 673358559 704393085 Methionine sulfoxide 2256830123 2386339603 2358839211 2725446801 2746599298 2870948492 3218287074 3175788605 3184222840 2824313157 2811707999 3170737686 methionine 417745146 383537536 458828825 530930768 493807736 510146524 640145587 612022776 621107832 540508664 492541936 462001014 methylthioadenosine 351692838 361103043 372364301 340715473 322074980 310762851 193553796 200101296 155392713 132432013 129879914 139584000 N-acetylaspartate 69696344.7 81710183.2 68566865.7 87051978.5 87155999.1 73804382.5 55923988.3 59223567.1 62710978.1 95571334 103610580.1 116420807.9 N-acetylglutamate 64046634 67474151 65712758 53553513 52753960 48599798 34354759 35606180 43766828 19839119 17384350 20950295 N-acetylphenylalanine 1406602.9 1250312.3 1314523.3 1011715.3 976298.6 1111549.3 783813.8 706720.9 647647.8 729161.8 825169.5 647704.5 NAD 715105861 693744032 736985034 623628700 641534937 601270701 501140555 540797676 580778501 397907708 410492506 408861953 NADP 19429278 23169320 21555186 17659877 18397685 15813744 17494174 17036685 19308324 13918592 16734297 13239117 niacinamide 90073800 100351828 88342396 103956842 113156252 136447784 106026962 106067920 87973572 103469247 115690066 112813804 O-phosphoethanolamine 1830665.5 1753784.4 1655657.3 1797322.4 2296185.7 2196604.8 7289968.4 6807560.1 7816992.4 16302260.4 13526725.3 14580185.2 ornithine 18730218 16168591 16986282 20358204 16084974 17203409 43586103 38689387 47231178 28272476 26975918 31875838 orotate 9211606 9224594 9198635 2388734 3344224 2514079 55791439 62890143 66280101 2050330 2065545 1927241 palmitate 167055970 112946014 99672923 97910306 91950949 95643204 81597892 82439903 61276196 93441600 78378150 102576377 Pantothenate 775096056 791732111 739824190 712377825 684783986 690274835 1444288507 1482821168 951554396 740413499 729254116 734428711 phenylalanine 172115669 165464873 173175494 214134682 189083069 205443351 265084860 272393751 197884019 237959438 213875859 231256932 phosphocholine 39297232306 36204598365 37511246884 34078661428 37971338153 36382242082 49502250038 50684100399 52636421427 18059763012 18241887105 19285475757 phosphocreatine 16651111 16805087 17259472 15665745 16772299 15737812 17799453 18934592 20080049 24611856 26323882 23557512 phosphoenolpyruvate 42851833 43205665 42668437 33596412 32850650 35009314 9958325 10086459 12929221 19347435 21012769 18448905 proline 18106132157 17813216289 18013183116 18422419413 17812069788 18672847109 19142055679 18563094472 17341911503 16218359441 16708475483 16842065004 propionylcarnitine 83405518 82821993 84525349 81586842 85223886 80090047 37698347 41324904 31543144 81330316 80871070 75889918 PRPP 10509048 12327018 12249589 69569836 79300057 76667153 3448540 3428135 4973192 78024953 79208797 78829915 Pyridoxine 183071673 176240966 169277434 172197565 152624160 180065549 243955202 232749652 166199474 164029796 180379634 147683770 Ribose-1-phosphate 3005591 2883708 3198502 12702954 12212354 12073722 1253555 1095396 1571403 16464234 17266278 17267470 Ribose_Ribulose-5-phosphate-3Me-Lys 17095150 23393378 20415540 14986112 16233917 15258655 9425953 10343701 9645720 15480257 16215728 16377705 S-Adenosyl-L-Homocysteine 17033310 18246590 17727538 15406658 15567161 15096422 44178544 45741107 49426877 10941193 11116836 11202048 S-Adenosyl-L-methionine 458922954 462342140 376263130 467871071 449172356 385733950 260682136 271234482 272838660 388850080 380441851 440216084 Sedoheptulose-7-phosphate 7662987 6835214 6589023 5848055 5279388 5699829 19395661 19229645 18718049 3350597 3578623 3457455 serine 47111371 38629641 42416908 147933454 151522053 152004510 101750926 105247091 117520386 272706746 236643229 262233717 succinate 88390682 88013208 83305020 50348206 47014119 43486221 21356194 21478568 19953732 37332647 34918846 36892741 succinyl-CoA 5477777 5142575 6406292 4847995 7055381 5953019 4448668 5666886 7162577 2823003 4028604 2831989 taurine 13584234 13077001 14245504 14195635 14408709 14621088 22000953 24663522 24745749 21051284 19119604 20643573 threonine 711451982 689659297 698412676 834207522 880388271 861615019 1194175548 1245119408 1292597520 921937043 973233575 965308632 tryptophan 265961633 253528841 284963758 307133847 291735339 318524832 383367036 365825505 331100051 338809340 324741377 309421193 tyrosine 1075186048 1042579499 1116566576 1235923772 1115421797 1167188396 1269467970 1240453212 1342951394 1130815738 1043035048 1147743114 UDP-GlcNAc 249357371 245356999 243551989 178833389 182625097 172558729 23973691 24190205 25977504 100346667 102151648 102505101 UDP 14593428 16492075 15428086 18320067 19731596 16261160 2207866 2247223 2979349 11957898 12860714 13701029 UMP 2870120.5 3490300.3 3112232.4 3243636.6 3337144.5 3173961.7 467476.2 414077 302438.2 3218723.7 3316893.8 3415491 Ureidosuccinic acid 9726344 10164810 10781564 4955859 5698248 5047612 20566094 22090683 21260221 4058442 3822652 4209516 UTP 321213932 311268175 432064452 371646479 525147431 521799176 66187820 72624550 90732838 229166137 263416528 236002940 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