#METABOLOMICS WORKBENCH juliehaines_20210702_093259 DATATRACK_ID:2737 STUDY_ID:ST001864 ANALYSIS_ID:AN003021 PROJECT_ID:PR001177 VERSION 1 CREATED_ON July 17, 2021, 11:15 am #PROJECT PR:PROJECT_TITLE Targeting host glycolysis as a strategy for antimalarial development PR:PROJECT_SUMMARY Glycolysis controls cellular energy, redox balance, and biosynthesis. PR:PROJECT_SUMMARY Antiglycolytic therapies are under investigation for treatment of obesity, PR:PROJECT_SUMMARY cancer, aging, autoimmunity, and microbial diseases. Interrupting glycolysis is PR:PROJECT_SUMMARY highly valued as a therapeutic strategy, because glycolytic disruption is PR:PROJECT_SUMMARY generally tolerated in mammals. Unfortunately, anemia is a known dose-limiting PR:PROJECT_SUMMARY side effect of these inhibitors and presents a major caveat to development of PR:PROJECT_SUMMARY antiglycolytic therapies. We developed specific inhibitors of enolase – a PR:PROJECT_SUMMARY critical enzyme in glycolysis – and validated their metabolic and cellular PR:PROJECT_SUMMARY effects on human erythrocytes. Enolase inhibition increases erythrocyte PR:PROJECT_SUMMARY susceptibility to oxidative damage and induces rapid and premature erythrocyte PR:PROJECT_SUMMARY senescence, rather than direct hemolysis. We apply our model of red cell PR:PROJECT_SUMMARY toxicity to address questions regarding erythrocyte glycolytic disruption in the PR:PROJECT_SUMMARY context of Plasmodium falciparum malaria pathogenesis. Our study provides a PR:PROJECT_SUMMARY framework for understanding red blood cell homeostasis under normal and disease PR:PROJECT_SUMMARY states and clarifies the importance of erythrocyte reductive capacity in malaria PR:PROJECT_SUMMARY parasite growth. PR:INSTITUTE University of Colorado Anschutz Medical Campus PR:LAST_NAME Haines PR:FIRST_NAME Julie PR:ADDRESS 12801 E 17th Ave, Room 1303, Aurora, Colorado, 80045, USA PR:EMAIL julie.haines@cuanschutz.edu PR:PHONE 3037243339 #STUDY ST:STUDY_TITLE Targeting host glycolysis as a strategy for antimalarial development ST:STUDY_SUMMARY Glycolysis controls cellular energy, redox balance, and biosynthesis. ST:STUDY_SUMMARY Antiglycolytic therapies are under investigation for treatment of obesity, ST:STUDY_SUMMARY cancer, aging, autoimmunity, and microbial diseases. Interrupting glycolysis is ST:STUDY_SUMMARY highly valued as a therapeutic strategy, because glycolytic disruption is ST:STUDY_SUMMARY generally tolerated in mammals. Unfortunately, anemia is a known dose-limiting ST:STUDY_SUMMARY side effect of these inhibitors and presents a major caveat to development of ST:STUDY_SUMMARY antiglycolytic therapies. We developed specific inhibitors of enolase – a ST:STUDY_SUMMARY critical enzyme in glycolysis – and validated their metabolic and cellular ST:STUDY_SUMMARY effects on human erythrocytes. Enolase inhibition increases erythrocyte ST:STUDY_SUMMARY susceptibility to oxidative damage and induces rapid and premature erythrocyte ST:STUDY_SUMMARY senescence, rather than direct hemolysis. We apply our model of red cell ST:STUDY_SUMMARY toxicity to address questions regarding erythrocyte glycolytic disruption in the ST:STUDY_SUMMARY context of Plasmodium falciparum malaria pathogenesis. Our study provides a ST:STUDY_SUMMARY framework for understanding red blood cell homeostasis under normal and disease ST:STUDY_SUMMARY states and clarifies the importance of erythrocyte reductive capacity in malaria ST:STUDY_SUMMARY parasite growth. ST:INSTITUTE University of Colorado Anschutz Medical Campus ST:LAST_NAME Haines ST:FIRST_NAME Julie ST:ADDRESS 12801 E 17th Ave, Room 1303, Aurora, Colorado, 80045, USA ST:EMAIL julie.haines@cuanschutz.edu ST:PHONE 3037243339 #SUBJECT SU:SUBJECT_TYPE Cultured cells SU:SUBJECT_SPECIES Homo sapiens SU:TAXONOMY_ID 9606 #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 - 2 group:Control, 0h RAW_FILE_NAME=2 SUBJECT_SAMPLE_FACTORS - 4 group:Control, 0h RAW_FILE_NAME=4 SUBJECT_SAMPLE_FACTORS - 6 group:Control, 0h RAW_FILE_NAME=6 SUBJECT_SAMPLE_FACTORS - 20 group:POMSF, 0h RAW_FILE_NAME=20 SUBJECT_SAMPLE_FACTORS - 22 group:POMSF, 0h RAW_FILE_NAME=22 SUBJECT_SAMPLE_FACTORS - 24 group:POMSF, 0h RAW_FILE_NAME=24 SUBJECT_SAMPLE_FACTORS - 26 group:POMHEX, 0h RAW_FILE_NAME=26 SUBJECT_SAMPLE_FACTORS - 28 group:POMHEX, 0h RAW_FILE_NAME=28 SUBJECT_SAMPLE_FACTORS - 30 group:POMHEX, 0h RAW_FILE_NAME=30 SUBJECT_SAMPLE_FACTORS - 32 group:Control, 0.5h RAW_FILE_NAME=32 SUBJECT_SAMPLE_FACTORS - 34 group:Control, 0.5h RAW_FILE_NAME=34 SUBJECT_SAMPLE_FACTORS - 36 group:Control, 0.5h RAW_FILE_NAME=36 SUBJECT_SAMPLE_FACTORS - 50 group:POMSF, 0.5h RAW_FILE_NAME=50 SUBJECT_SAMPLE_FACTORS - 52 group:POMSF, 0.5h RAW_FILE_NAME=52 SUBJECT_SAMPLE_FACTORS - 54 group:POMSF, 0.5h RAW_FILE_NAME=54 SUBJECT_SAMPLE_FACTORS - 56 group:POMHEX, 0.5h RAW_FILE_NAME=56 SUBJECT_SAMPLE_FACTORS - 58 group:POMHEX, 0.5h RAW_FILE_NAME=58 SUBJECT_SAMPLE_FACTORS - 60 group:POMHEX, 0.5h RAW_FILE_NAME=60 SUBJECT_SAMPLE_FACTORS - 62 group:Control, 1h RAW_FILE_NAME=62 SUBJECT_SAMPLE_FACTORS - 64 group:Control, 1h RAW_FILE_NAME=64 SUBJECT_SAMPLE_FACTORS - 66 group:Control, 1h RAW_FILE_NAME=66 SUBJECT_SAMPLE_FACTORS - 80 group:POMSF, 1h RAW_FILE_NAME=80 SUBJECT_SAMPLE_FACTORS - 82 group:POMSF, 1h RAW_FILE_NAME=82 SUBJECT_SAMPLE_FACTORS - 84 group:POMSF, 1h RAW_FILE_NAME=84 SUBJECT_SAMPLE_FACTORS - 86 group:POMHEX, 1h RAW_FILE_NAME=86 SUBJECT_SAMPLE_FACTORS - 88 group:POMHEX, 1h RAW_FILE_NAME=88 SUBJECT_SAMPLE_FACTORS - 90 group:POMHEX, 1h RAW_FILE_NAME=90 SUBJECT_SAMPLE_FACTORS - 122 group:Control, 6h RAW_FILE_NAME=122 SUBJECT_SAMPLE_FACTORS - 124 group:Control, 6h RAW_FILE_NAME=124 SUBJECT_SAMPLE_FACTORS - 126 group:Control, 6h RAW_FILE_NAME=126 SUBJECT_SAMPLE_FACTORS - 140 group:POMSF, 6H RAW_FILE_NAME=140 SUBJECT_SAMPLE_FACTORS - 142 group:POMSF, 6H RAW_FILE_NAME=142 SUBJECT_SAMPLE_FACTORS - 144 group:POMSF, 6H RAW_FILE_NAME=144 SUBJECT_SAMPLE_FACTORS - 146 group:POMHEX, 6H RAW_FILE_NAME=146 SUBJECT_SAMPLE_FACTORS - 148 group:POMHEX, 6H RAW_FILE_NAME=148 SUBJECT_SAMPLE_FACTORS - 150 group:POMHEX, 6H RAW_FILE_NAME=150 #COLLECTION CO:COLLECTION_SUMMARY Banked blood (Saint Louis Children’s Hospital) and fresh blood [healthy CO:COLLECTION_SUMMARY donors] was washed and stored at 50% hematocrit at 4ºC up to 1 month past the CO:COLLECTION_SUMMARY clinical expiration date and collection date, respectively. Parasite strains CO:COLLECTION_SUMMARY provided by the Malaria Research and Reference Reagent Resource Center (MR4) as CO:COLLECTION_SUMMARY follows: 3D7 (MRA-102); K1 (MRA-159); D10 (MRA-201); and IPC-5202 (MRA-1240). CO:COLLECTION_SUMMARY Unless indicated, erythrocytes were cultured as previously described at 2% CO:COLLECTION_SUMMARY hematocrit in complete media (incomplete media supplemented with 27mM sodium CO:COLLECTION_SUMMARY bicarbonate, 11mM glucose, 5mM HEPES, 1mM sodium pyruvate, 0.37mM hypoxanthine, CO:COLLECTION_SUMMARY 0.01mM thymidine, 10ug/mL gentamicin, and 0.5% Albumax (Thermo-Fisher) under 5% CO:COLLECTION_SUMMARY O2/5% CO2/90% N2 atmosphere at 37ºC (71). CO:SAMPLE_TYPE Blood (whole) #TREATMENT TR:TREATMENT_SUMMARY Freshly collected erythrocytes were washed, resuspended to 10% hematocrit in TR:TREATMENT_SUMMARY buffer [25mM HEPES (pH 7.4), 120mM NaCl, 5.4mM KCl, 1.8mM CaCl2, and 1mM TR:TREATMENT_SUMMARY NaH2PO4], incubated for 1 hour at 37ºC, then centrifuged at 2000xg for 5 TR:TREATMENT_SUMMARY minutes at 4ºC, resuspended in fresh wash buffer to 30-40% hematocrit, and TR:TREATMENT_SUMMARY split into 210uL aliquots of packed erythrocytes. Erythrocytes were resuspended TR:TREATMENT_SUMMARY in 253uL of RPMI containing 11.9mM D-[1,2,3-13C] glucose (Millipore-Sigma) and TR:TREATMENT_SUMMARY either treated with POM-SF or POM-HEX at five times the EC50s for MetHb TR:TREATMENT_SUMMARY formation, or an untreated solvent control with the balance volume to 550uL TR:TREATMENT_SUMMARY using washing buffer. All samples were incubated at 37ºC shaking at 500 RPM. TR:TREATMENT_SUMMARY Triplicate samples of supernatants and packed erythrocytes were collected at 0, TR:TREATMENT_SUMMARY 0.5, 1, and 6-hour intervals and snap-frozen in liquid N2. #SAMPLEPREP SP:SAMPLEPREP_SUMMARY RBC and media samples were extracted at 1:10 and 1:20 dilutions, respectively, SP:SAMPLEPREP_SUMMARY in ice-cold 5:3:2 MeOH:MeCN:H2O v/v/v, with vigorous vortexing at 4 degrees C SP:SAMPLEPREP_SUMMARY for 30 min. The extraction buffer was supplemented with 40uM 3-13C lactate to SP:SAMPLEPREP_SUMMARY allow absolute quantification of lactate isotopologues, including lactate SP:SAMPLEPREP_SUMMARY generated via glycolysis ([1,2,3-13C3] enriched) vs. the pentose phosphate SP:SAMPLEPREP_SUMMARY pathway ([2,3-13C2] enriched). Supernatants were clarified via centrifugation SP:SAMPLEPREP_SUMMARY (10,000 g, 10 min, 4 C) then analyzed by UHPLC-MS. #CHROMATOGRAPHY CH:CHROMATOGRAPHY_SUMMARY 10 μl of RBC extracts were injected and separated through a 3 min isocratic CH:CHROMATOGRAPHY_SUMMARY elution on a Kinetex XB-C18 column (150 × 2.1 mm i.d., 1.7 μm particle size CH:CHROMATOGRAPHY_SUMMARY – Phenomenex, Torrance, CA, USA) at 250 μl/min (mobile phase: 5% CH:CHROMATOGRAPHY_SUMMARY acetonitrile, 95% 18 mΩ H2O, 0.1% formic acid; column temperature: 25°C). CH:CHROMATOGRAPHY_TYPE Reversed phase CH:INSTRUMENT_NAME Vanquish CH:COLUMN_NAME Kinetex XB-C18 2.1 x 150 mm, 1.7 um CH:FLOW_GRADIENT isocratic CH:FLOW_RATE 250 ul/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 NEGATIVE MS:MS_COMMENTS The UHPLC system was coupled online with a QExactive mass spectrometer (Thermo, MS:MS_COMMENTS San Jose, CA, USA), scanning in Full MS mode (2 μscans) at 70,000 resolution MS:MS_COMMENTS from 60-900 m/z, with 4 kV spray voltage, 15 sheath gas and 5 auxiliary gas, MS:MS_COMMENTS operated in positive ion mode. Calibration was performed before each analysis MS:MS_COMMENTS using a positive calibration mix (Piercenet – Thermo Fisher, Rockford, IL, MS:MS_COMMENTS USA). Limits of detection (LOD) were characterized by determining the smallest MS:MS_COMMENTS injected amino acid amount required to provide a signal to noise (S/N) ratio MS:MS_COMMENTS greater than three using < 5 ppm error on the accurate intact mass. Based on a MS:MS_COMMENTS conservative definition for Limit of Quantitation (LOQ), these values were MS:MS_COMMENTS calculated to be three fold higher than determined LODs. MS data acquired from MS:MS_COMMENTS the QExactive was converted from .raw file format to.mzXML format using MS:MS_COMMENTS MassMatrix (Cleveland, OH, USA). Amino acid assignments were performed using MS:MS_COMMENTS MAVEN (Princeton, NJ, USA). The MAVEN software platform provides tools for peak MS:MS_COMMENTS picking, feature detection and metabolite assignment against the KEGG pathway MS:MS_COMMENTS database. Assignments were further confirmed using a process for chemical MS:MS_COMMENTS formula determination using isotopic patterns and accurate intact mass (Clasquin MS:MS_COMMENTS et al. 2012). Analyte retention times were confirmed by comparison with external MS:MS_COMMENTS standard retention times, as indicated above. #MS_METABOLITE_DATA MS_METABOLITE_DATA:UNITS peak area MS_METABOLITE_DATA_START Samples 2 4 6 20 22 24 26 28 30 32 34 36 50 52 54 56 58 60 62 64 66 80 82 84 86 88 90 122 124 126 140 142 144 146 148 150 Factors group:Control, 0h group:Control, 0h group:Control, 0h group:POMSF, 0h group:POMSF, 0h group:POMSF, 0h group:POMHEX, 0h group:POMHEX, 0h group:POMHEX, 0h group:Control, 0.5h group:Control, 0.5h group:Control, 0.5h group:POMSF, 0.5h group:POMSF, 0.5h group:POMSF, 0.5h group:POMHEX, 0.5h group:POMHEX, 0.5h group:POMHEX, 0.5h group:Control, 1h group:Control, 1h group:Control, 1h group:POMSF, 1h group:POMSF, 1h group:POMSF, 1h group:POMHEX, 1h group:POMHEX, 1h group:POMHEX, 1h group:Control, 6h group:Control, 6h group:Control, 6h group:POMSF, 6H group:POMSF, 6H group:POMSF, 6H group:POMHEX, 6H group:POMHEX, 6H group:POMHEX, 6H ATP 4996 0 9360 13554 11596 9689 7617 7582 9000 1247 1649 5959 6368 8430 5728 4242 2548 4822 5508 8327 27959 15715 28195 22627 13232 21058 34319 25833 30957 33908 15788 98414 96266 106057 45071 28578 Adenine 155573 157677 133462 141405 142095 151346 130928 138291 132324 143706 212208 169070 192117 176841 166973 153847 160871 162725 133971 116557 173002 134362 122348 152050 175377 163003 161234 108081 102219 121249 77539 123226 112136 110300 113492 102509 dCMP 41428 32084 34222 30749 21603 22076 26639 25745 19130 30550 20178 29330 18655 18411 17611 23390 23674 17827 15537 13201 18643 15055 14247 11478 6467 14178 8648 13205 13629 17155 0 11924 15139 16190 16560 13481 dTMP 1334314 1515622 1791353 1280553 1665920 976289 1471962 1260596 964826 1533592 1430963 1626038 1286060 841648 1179406 979306 1146766 1172310 1257445 1172905 1485518 1277548 1273799 1219071 695331 1438131 566101 1223864 1123150 1213312 26130 1278508 708960 1096031 1203518 963416 Thymidine 20511 25173 16315 25013 13104 11486 19296 13921 12769 14437 8465 8215 13356 11735 18688 10088 19440 13705 4858 10220 4141 11113 12353 8294 11174 12569 8727 6294 5468 5302 40305 7722 7984 12435 9431 3227 UDP 348921 446596 539951 645076 464336 160294 346963 430996 243498 317194 217844 399915 208770 117556 138145 169248 266087 202840 208365 131228 168142 225038 189369 139634 50667 141860 28785 96638 78814 148355 0 131941 72660 88316 99351 81831 IMP 22762 29766 16589 77065 27326 21692 20667 41288 24207 8239 56524 47030 91275 0 41075 0 171307 80085 48877 83228 95930 157653 98045 195850 78082 91525 62250 32992 29168 31724 12657 82577 57589 45104 37354 46106 Inosine 134173 162015 141077 94707 113497 120163 122982 118580 108886 113669 113045 107883 90743 109477 115346 108688 69468 73526 87063 60622 77190 67052 62858 66298 62193 54415 60496 64322 65688 66193 1210 45887 55018 68394 67321 41971 Hypoxanthine 111421 140665 120873 146510 128974 132966 122893 84994 106533 9362 7377 4940 65281 43644 36625 40669 46841 43745 6407 4546 0 32061 16551 8152 6658 17124 16851 0 0 2746 7616 4258 3536 7139 5927 0 (S)(+)-Allantoin 43815 38651 35316 39585 41936 47795 39990 45010 44303 39042 33894 40028 43474 35833 0 21535 0 29330 33053 16010 17986 17194 15666 26213 18326 17007 14842 27402 13742 14105 1758 8427 0 7398 8256 9045 Urate 36476 48321 55883 49370 41439 37158 47909 46201 32769 25868 24950 41171 35128 27233 26812 33989 37853 32360 36205 22342 35904 32671 29437 24566 23145 28011 21250 28767 30877 25574 13873 23021 20133 35801 28389 20275 5-6-Dihydrothymine 360466 394442 347899 313538 340962 285222 337339 323393 269611 343159 328464 298534 310331 280349 287173 314187 295256 281872 287456 293798 304448 250033 277099 296120 256585 302573 184726 290656 225975 269056 74230 303273 279076 280202 287318 228016 Pyridoxal 73730 74291 52514 58277 44603 62797 50617 48081 56808 82221 81026 86184 79482 87576 84178 73430 76145 76359 106200 58969 39353 58439 57464 69192 52477 82276 75860 30076 35575 63209 37635 43949 53550 42611 52387 76411 UDP-glucose 88336 223859 165693 112270 115043 91853 80515 61533 63116 35780 24773 34517 30929 35523 41470 38616 21425 29597 17736 765 19226 23808 17207 9179 5615 10752 6045 12679 11077 13294 0 8804 10280 9760 1906 5747 Diphosphate 1478 1530 1079 6505 2623 712 2292 2814 1297 2032 2780 5546 3259 1342 1516 3535 8585 2398 6550 29905 15760 29393 33142 17665 13961 38516 17282 82077 76985 49039 0 29865 21583 28388 35225 38553 D-Glucose 8049242 8472276 8804735 6823206 7468972 6933306 7033582 7963980 6067010 8301542 6913984 6934990 6069978 5639556 6251898 6036718 6913766 5920758 6111762 4492928 6363206 5285640 5041350 5139284 4527030 6779172 3725381 4732162 3750000 4090909 33887 4317652 3638352 3987520 4573066 4900162 Pyruvate 1565338 1718396 1603873 1181566 1327064 1202172 821317 791458 705806 1699000 1945312 1743200 667261 732488 675195 585828 663681 639012 1668893 1351845 2003297 901194 795157 815340 538943 710162 433353 4652690 4951628 5285586 86796 2040818 2168570 2166179 2174124 1352580 Lactate 9622396 11799080 9511139 7244318 8300388 8235258 7439496 7840298 7124236 9394389 10481420 9852840 9038235 8729325 8568688 8534571 7591390 8320248 10001220 8592491 10247760 9218460 8764350 8977471 7952554 9123597 7544254 21144780 20552450 22207770 968065 16937080 17591160 17511190 20746070 14291810 Mannitol 171232 195207 198243 86740 121680 145357 100666 111236 122377 182034 169250 152372 147786 143065 143502 133682 114501 142141 147146 93285 151669 101188 95280 138356 125466 153741 102012 134067 116475 117560 6477 185881 178053 182539 122386 151302 D-Ribose 221744 283709 242035 155123 241413 238444 242376 207795 199883 189439 201336 191470 175120 210014 184323 189897 142313 170950 177000 89873 120852 163968 119055 118158 122465 196118 118315 97093 103247 113181 13676 142838 111222 110664 89668 84312 Citrate 31621 41872 37074 36586 33107 36355 27297 31864 25922 31208 28141 24151 26317 32315 35649 42063 33807 34275 35221 25419 40905 43514 38232 32946 18897 21015 14610 19410 24445 31052 1565 17707 24182 27556 19124 15734 Succinate 107275 122919 131639 97915 114367 109646 110490 106485 108234 90442 95200 90086 95711 95081 94486 71388 94417 66649 105679 75347 105366 74494 84779 57426 61319 99444 57930 80920 75758 78864 14015 65505 45495 38729 59728 70477 Fumarate 167282 247653 184237 165981 193119 168237 169621 153197 129130 126001 113341 139797 107818 133248 127457 124078 95232 109576 111601 71780 97191 104334 88605 88660 81639 80816 77780 76709 77443 100728 3336 78083 105708 89979 78435 71857 Malate 613170 1009831 790564 552497 572824 531395 407404 312929 302501 299652 223267 295276 184141 215487 229663 209452 130094 161757 173294 30964 148377 154806 107137 99103 67598 61664 62482 142749 249965 277224 3138 106843 155909 161383 93877 76080 Oxaloacetate 36859 53328 66037 69654 60109 49843 48215 53249 43526 63792 42493 43638 40952 38219 35270 39335 56018 50529 33858 67206 48295 56926 54229 43825 32915 54891 30333 36448 38018 60288 1482 34360 36028 37179 44257 34732 2-Hydroxyglutarate/Citramalate 215496 306628 265936 213633 239758 204358 186658 144069 135730 141634 109826 130836 87823 97157 118054 105598 67005 98059 80538 22646 80529 78593 56900 56112 44498 47580 42362 83746 109789 114557 8757 58264 80827 78391 57510 56561 Sedoheptulose 1-phosphate 38854 56618 57220 52308 62525 48226 45258 41432 29182 68834 16192 40269 15618 16761 17962 15709 13267 12300 37363 9523 29471 15864 19329 8805 2813 3839 2378 13436 40826 43272 0 16645 19154 23871 13211 9548 alpha-D-Ribose 1-phosphate 111310 191964 156687 148000 143781 126163 159576 142880 110714 103430 63058 95524 108376 88361 107114 149449 116898 85648 54967 22223 51603 63920 66663 43209 22899 47769 29059 13321 21359 42552 0 28916 37545 43368 29154 19657 Glutathione disulfide 301983 1947697 1561140 384133 678363 493058 406596 298663 343854 373481 123372 291315 136465 252054 391218 233604 146227 254886 164540 28584 168027 147918 96399 95020 74458 75687 54062 81399 102098 254798 0 162029 364292 299940 93797 78062 5-Oxoproline 1236168 1328848 1327924 1056072 1185607 1047367 1187766 1085712 990520 990364 1046222 1045592 853777 965437 967155 1014498 827034 880442 904716 705176 863637 826569 757396 713850 598692 610183 620858 979178 868937 1007876 76549 826852 866679 882702 1091526 756716 S-Glutathionyl-L-cysteine 72060 208247 234832 180128 227508 91585 167972 157413 87481 206741 61861 200555 64369 85841 221486 160810 105196 176793 131027 103137 141829 118298 85369 76873 55712 60183 18246 52669 50022 110848 0 118495 130208 223729 79988 69172 Ascorbate 126826 149868 126379 126748 155056 106776 103111 139434 105388 110010 99936 103326 92980 68466 107504 93841 110374 113235 104656 76260 94174 116813 85287 79978 63047 107804 49715 83542 77844 63670 317 84874 70969 76441 92095 92546 Dehydroascorbate 8273 21070 18182 19716 12353 6281 6656 12553 9161 8125 21827 35116 17831 9280 17741 15529 21997 17188 19330 47770 45972 63177 58651 41200 40325 37836 25573 38724 39007 28209 0 37271 18170 35744 43935 46486 gamma-L-Glutamyl-L-cysteine 20176 39331 48139 48340 56078 6443 28076 36404 25106 24025 17398 49346 13905 23920 23648 22580 48191 47841 37462 52950 32048 39576 57582 18983 14260 36786 6674 42432 29269 48472 4078 26751 32416 41960 30269 39606 gamma-L-Glutamyl-D-alanine 31967 47711 39005 24083 30816 25178 24809 21900 18494 22816 39960 35119 39928 30948 33098 36051 29453 28599 34686 37467 43269 42611 30689 31168 18074 17373 16996 31381 41244 31786 386 30610 33717 46980 44892 27355 L-Homoserine 205133 267673 254816 210612 232343 146695 184529 218624 131480 270908 236140 277175 213750 201289 222570 190540 239348 186083 237748 194552 268161 234381 210010 234656 175727 251608 157859 266690 238016 272500 15991 274579 250385 261931 331460 239491 N-Acetylneuraminate 33653 46086 42040 22774 33173 37837 30533 32686 22660 31564 36131 31154 28389 27554 26329 30228 18886 19115 29921 23553 26201 22465 16837 23762 19780 13191 16179 21032 17837 26053 0 13284 14987 16611 14571 8129 N-Glycoloyl-neuraminate 283419 286809 280175 222227 226544 172472 201581 251648 161112 219192 177513 189257 179704 143185 172157 166185 174685 161855 144337 125310 161366 122808 150786 132437 79688 141624 56781 99865 69489 104702 0 102341 73677 76429 86787 114034 1-4-beta-D-Xylan 345091 389543 330978 249629 293792 253584 275136 319138 260290 352327 283656 290819 242024 255437 250744 240162 274756 163545 250940 204358 234551 197257 192046 208801 176262 260414 154142 186214 178694 157545 24362 172097 176746 164417 139784 193451 UDP-N-acetyl-D-glucosamine 26431 70700 55685 36027 35042 34007 27689 21273 21326 15002 13455 23183 15481 20115 22577 18482 11419 18515 13816 1211 10150 12416 7594 4655 2550 3513 2644 6296 5242 8947 0 5998 10066 8060 2610 5407 CMP-N-acetylneuraminate 29122 199798 147466 34917 64026 44067 32195 26253 31739 37785 10989 28526 13761 22580 34483 21389 14578 21922 16589 1147 16406 16667 8288 7764 6053 4632 2615 4991 8143 21757 0 16000 33043 25469 6907 6080 N-Acetylornithine 89232 88008 65568 53555 73979 59129 55452 46080 54560 47201 61527 54732 69939 57083 55264 64671 47320 71112 53922 56537 62476 59630 39510 59002 57347 47059 45993 48007 56533 54069 8476 60249 54272 75937 58952 53290 trans-4-Hydroxy-L-proline 183162 195253 225280 213621 184573 121540 177353 161177 142903 208127 209166 255842 178809 164663 179851 167920 218492 176852 236580 181868 243772 163277 197237 207635 136630 240224 116456 275243 241271 245511 22561 253389 235411 250649 228558 260776 Aminofructose 6-phosphate 19631 20309 11479 12034 11806 21543 18597 16622 17006 15844 12482 16483 14802 13737 15474 17174 8117 13775 14426 6841 10466 10390 9959 8382 5713 4205 9016 7123 3403 7288 0 6173 11205 11227 9740 5644 L-Cysteate 42974 65947 62435 66734 75839 39334 63094 64127 35545 71220 41244 64966 37692 32543 53061 53884 38835 42225 55313 25552 38332 50158 40542 29903 25480 35245 21621 11616 17169 23141 1405 25132 25291 30239 21772 27158 Dopamine 48701 46623 39363 21431 35210 52477 33487 29975 33348 34497 34762 13637 29612 38098 26510 37121 22752 32129 26960 22427 33355 12082 26271 35929 40156 16259 23188 22839 19198 15124 2470 25678 28945 32529 21109 15755 Glycerol 3-phosphate 373365 496572 391808 365360 365626 421151 332369 266283 301584 199166 190095 192774 213348 222148 205232 190760 134870 149619 120352 51814 100732 130750 92245 97690 87341 86538 66749 34894 40798 51060 0 51986 71800 63845 34155 26504 Ethanolamine phosphate 37629 37223 45618 46358 45245 23140 32177 32187 37322 25432 26901 42529 29686 29352 35482 38012 30593 36142 27977 31346 26238 34263 25281 24713 19911 20151 17590 20070 19209 22365 0 24185 20642 15459 22477 14459 sn-glycero-3-Phosphoethanolamine 711103 594829 446135 214755 338623 436427 253475 205003 228384 399247 341101 214926 325656 386942 342022 253808 241233 193776 271447 124935 198117 111303 123446 196050 221835 151091 219094 153266 129909 176081 1628 158709 270945 229094 174784 124901 6-Pyruvoyltetrahydropterin 225356 239723 231189 154897 174884 194763 200588 175850 171219 167125 180873 173959 142502 173212 173798 170591 119041 142080 132880 87999 105218 97522 98169 107623 110842 99634 106619 74254 67437 95376 0 72510 97518 99129 71573 61880 Peptide tryptophan 70818 81559 60723 45895 66967 67978 61796 65704 61270 59086 51776 46221 55188 59583 64665 57858 50255 51139 56146 15797 48875 47154 39628 44766 37438 27435 38689 17981 35341 47263 0 30520 30239 21124 26555 34606 Shikimate 3-phosphate 21738 30654 32588 22352 29411 23164 31739 22750 20710 21077 23915 26753 18257 20963 23095 18790 23965 21593 27665 16185 25986 18412 24146 19094 21318 20201 14500 21122 23574 21779 0 19652 19559 22539 21083 26057 D-glucono-1,5-lactone 845706 817854 729933 527816 759427 780222 734594 638626 633951 594519 623275 513560 520480 645187 570327 541817 439220 430989 455202 291958 368958 306159 320328 338017 336858 301216 347600 244763 253130 271906 12136 210166 285182 258142 264081 328091 D-Ribitol 5-phosphate 21278 24199 37273 19992 34781 20871 11754 27797 25464 31973 14425 38967 18882 26958 17769 20233 26702 23229 28838 21194 22422 20843 26674 23190 13604 37942 11051 19133 15142 32283 0 16167 16479 11543 23020 22473 Thioredoxin disulfide 48060 72131 82748 79979 122816 30298 85589 87405 64351 102875 40458 69774 44538 25341 35160 58481 15269 44581 37577 66535 122548 57174 102176 51392 34873 117137 21266 72827 44879 151084 0 80427 25084 22909 26926 27713 Pyridoxamine 28710 31207 31916 33066 30111 23647 34711 21117 41279 18131 46386 27529 43464 31469 39599 32477 29683 29518 35671 19911 21654 16790 15404 25820 12589 18140 8562 8507 6627 16896 19708 7815 6443 9673 8576 9605 L-Noradrenaline 57708 86686 116767 274149 173938 96155 209882 267461 153914 75700 54091 51565 52982 62593 58353 75090 132468 65868 49467 29011 31591 31608 37978 32028 32893 40392 28326 11199 14059 39210 16677 18065 31279 20156 27634 17048 Oxalosuccinate 211377 155524 166147 189940 187926 183549 180083 188758 174550 174174 161178 157768 159950 151344 130278 138552 139768 127396 123795 91417 114750 93656 103991 96323 98524 97232 100136 88545 92638 85083 37426 91942 102156 84881 89431 79975 MS_METABOLITE_DATA_END #METABOLITES METABOLITES_START metabolite_name CmpdID parent medRt ATP C00002 505.9883 1.13 Adenine C00147 134.0458 1.22 dCMP C00239 306.0491 1.57 dTMP C00364 321.0525 1.18 Thymidine C00214 241.0810 1.22 UDP C00015 402.9998 1.26 IMP C00130 347.0396 1.44 Inosine C00294 267.0721 1.12 Hypoxanthine C00262 135.0299 1.61 (S)(+)-Allantoin C02350 157.0345 1.42 Urate C00366 167.0213 1.14 5-6-Dihydrothymine C00906 127.0498 1.18 Pyridoxal C00250 166.0498 1.22 UDP-glucose C00029 565.0470 1.10 Diphosphate C00013 176.9349 1.45 D-Glucose C00031 179.0551 1.18 Pyruvate C00022 87.0071 1.18 Lactate C01432 89.0228 1.18 Mannitol C00392 181.0712 1.18 D-Ribose C00121 149.0447 1.12 Citrate C00158 191.0187 1.12 Succinate C00042 117.0177 1.18 Fumarate C00122 115.0021 1.12 Malate C00149 133.0127 1.11 Oxaloacetate C00036 131.0001 1.16 2-Hydroxyglutarate/Citramalate C02630 147.0287 1.11 Sedoheptulose 1-phosphate C06222 289.0329 1.12 alpha-D-Ribose 1-phosphate C00620 229.0112 1.12 Glutathione disulfide C00127 611.1442 1.09 5-Oxoproline C01879 128.0338 1.14 S-Glutathionyl-L-cysteine C05526 425.0803 1.11 Ascorbate C00072 175.0241 1.18 Dehydroascorbate C05422 173.0081 1.21 gamma-L-Glutamyl-L-cysteine C00669 249.0549 1.18 gamma-L-Glutamyl-D-alanine C03738 217.0822 1.12 L-Homoserine C00263 118.0494 1.19 N-Acetylneuraminate C00270 308.0986 1.12 N-Glycoloyl-neuraminate C03410 324.0901 1.18 1-4-beta-D-Xylan C02352 131.0334 1.19 UDP-N-acetyl-D-glucosamine C00043 606.0741 1.10 CMP-N-acetylneuraminate C00128 613.1414 1.09 N-Acetylornithine C00437 173.0920 1.21 trans-4-Hydroxy-L-proline C01157 130.0495 1.18 Aminofructose 6-phosphate C12214 258.0379 1.12 L-Cysteate C00506 167.9960 1.12 Dopamine C03758 152.0704 1.21 Glycerol 3-phosphate C00093 171.0053 1.12 Ethanolamine phosphate C00346 140.0105 1.13 sn-glycero-3-Phosphoethanolamine C01233 214.0479 1.21 6-Pyruvoyltetrahydropterin C03684 236.0771 1.12 Peptide tryptophan C02732 228.0800 1.21 Shikimate 3-phosphate C03175 253.0092 1.13 D-glucono-1,5-lactone C00198 177.0394 1.13 D-Ribitol 5-phosphate C01068 231.0273 1.17 Thioredoxin disulfide C00343 315.0184 1.18 Pyridoxamine C00534 167.0814 1.23 L-Noradrenaline C00547 168.0654 1.23 Oxalosuccinate C05379 189.0064 1.44 METABOLITES_END #END