#METABOLOMICS WORKBENCH kevin.cho@wustl.edu_20240409_091613 DATATRACK_ID:4766 STUDY_ID:ST003162 ANALYSIS_ID:AN005187 PROJECT_ID:PR001967 VERSION 1 CREATED_ON April 10, 2024, 6:57 pm #PROJECT PR:PROJECT_TITLE Loss of dihydroceramide desaturase drives neurodegeneration by disrupting PR:PROJECT_TITLE endoplasmic reticulum and lipid droplet homeostasis in glial cells PR:PROJECT_TYPE Untargeted Metabolomics and Lipidomics PR:PROJECT_SUMMARY Dihydroceramide desaturases convert dihydroceramides to ceramides, the PR:PROJECT_SUMMARY precursors of all complex sphingolipids. Reduction of DEGS1 dihydroceramide PR:PROJECT_SUMMARY desaturase function causes pediatric neurodegenerative disorder hypomyelinating PR:PROJECT_SUMMARY leukodystrophy-18 (HLD-18). We discovered that infertile crescent (ifc), the PR:PROJECT_SUMMARY Drosophila DEGS1 homolog, is expressed primarily in glial cells to promote CNS PR:PROJECT_SUMMARY development by guarding against neurodegeneration. Loss of ifc causes massive PR:PROJECT_SUMMARY dihydroceramide accumulation and severe morphological defects in cortex glia, PR:PROJECT_SUMMARY including endoplasmic reticulum (ER) expansion, failure of neuronal PR:PROJECT_SUMMARY ensheathment, and lipid droplet depletion. RNAi knockdown of the upstream PR:PROJECT_SUMMARY ceramide synthase schlank in glia of ifc mutants rescues ER expansion, PR:PROJECT_SUMMARY suggesting dihydroceramide accumulation in the ER drives this phenotype. RNAi PR:PROJECT_SUMMARY knockdown of ifc in glia but not neurons drives neuronal cell death, suggesting PR:PROJECT_SUMMARY that ifc function in glia promotes neuronal survival. Our work identifies glia PR:PROJECT_SUMMARY as the primary site of disease progression in HLD-18 and may inform on juvenile PR:PROJECT_SUMMARY forms of ALS, which also feature elevated dihydroceramide levels. PR:INSTITUTE Washington University in St. Louis PR:DEPARTMENT Genetics, Medicine, Chemistry PR:LABORATORY Skeath and Patti Laboratories PR:LAST_NAME Cho PR:FIRST_NAME Kevin PR:ADDRESS 1 Brookings Drive, Campus Box 1134, St. Louis, MO, 63130, USA PR:EMAIL kevin.cho@wustl.edu PR:PHONE 314-935-8813 #STUDY ST:STUDY_TITLE Loss of dihydroceramide desaturase drives neurodegeneration by disrupting ST:STUDY_TITLE endoplasmic reticulum and lipid droplet homeostasis in glial cells ST:STUDY_TYPE Untargeted Metabolomics & Lipidomics ST:STUDY_SUMMARY Dihydroceramide desaturases convert dihydroceramides to ceramides, the ST:STUDY_SUMMARY precursors of all complex sphingolipids. Reduction of DEGS1 dihydroceramide ST:STUDY_SUMMARY desaturase function causes pediatric neurodegenerative disorder hypomyelinating ST:STUDY_SUMMARY leukodystrophy-18 (HLD-18). We discovered that infertile crescent (ifc), the ST:STUDY_SUMMARY Drosophila DEGS1 homolog, is expressed primarily in glial cells to promote CNS ST:STUDY_SUMMARY development by guarding against neurodegeneration. Loss of ifc causes massive ST:STUDY_SUMMARY dihydroceramide accumulation and severe morphological defects in cortex glia, ST:STUDY_SUMMARY including endoplasmic reticulum (ER) expansion, failure of neuronal ST:STUDY_SUMMARY ensheathment, and lipid droplet depletion. RNAi knockdown of the upstream ST:STUDY_SUMMARY ceramide synthase schlank in glia of ifc mutants rescues ER expansion, ST:STUDY_SUMMARY suggesting dihydroceramide accumulation in the ER drives this phenotype. RNAi ST:STUDY_SUMMARY knockdown of ifc in glia but not neurons drives neuronal cell death, suggesting ST:STUDY_SUMMARY that ifc function in glia promotes neuronal survival. Our work identifies glia ST:STUDY_SUMMARY as the primary site of disease progression in HLD-18 and may inform on juvenile ST:STUDY_SUMMARY forms of ALS, which also feature elevated dihydroceramide levels. ST:INSTITUTE Washington University in St. Louis ST:DEPARTMENT Genetics, Medicine, Chemistry ST:LABORATORY Skeath and Patti Laboratories ST:LAST_NAME Cho ST:FIRST_NAME Kevin ST:ADDRESS 1 Brookings Drive, Campus Box 1134, St. Louis, MO, 63130, USA ST:EMAIL kevin.cho@wustl.edu ST:PHONE 314-935-8813 #SUBJECT SU:SUBJECT_TYPE Insect SU:SUBJECT_SPECIES Drosophila melanogaster SU:TAXONOMY_ID 7227 #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 - WL_RPLC_POS_ifc_1 Genotype:Knockout | Sample source:Whole Larvae Chromatography=RPLC; Polarity=Positive; RAW_FILE_NAME(Raw file name)=WL_RPLC_POS_ifc_1.mzML SUBJECT_SAMPLE_FACTORS - WL_RPLC_POS_ifc_2 Genotype:Knockout | Sample source:Whole Larvae Chromatography=RPLC; Polarity=Positive; RAW_FILE_NAME(Raw file name)=WL_RPLC_POS_ifc_2.mzML SUBJECT_SAMPLE_FACTORS - WL_RPLC_POS_ifc_3 Genotype:Knockout | Sample source:Whole Larvae Chromatography=RPLC; Polarity=Positive; RAW_FILE_NAME(Raw file name)=WL_RPLC_POS_ifc_3.mzML SUBJECT_SAMPLE_FACTORS - WL_RPLC_POS_ifc_4 Genotype:Knockout | Sample source:Whole Larvae Chromatography=RPLC; Polarity=Positive; RAW_FILE_NAME(Raw file name)=WL_RPLC_POS_ifc_4.mzML SUBJECT_SAMPLE_FACTORS - WL_RPLC_POS_ifc_5 Genotype:Knockout | Sample source:Whole Larvae Chromatography=RPLC; Polarity=Positive; RAW_FILE_NAME(Raw file name)=WL_RPLC_POS_ifc_5.mzML SUBJECT_SAMPLE_FACTORS - WL_RPLC_POS_WT_1 Genotype:Wild-type | Sample source:Whole Larvae Chromatography=RPLC; Polarity=Positive; RAW_FILE_NAME(Raw file name)=WL_RPLC_POS_WT_1.mzML SUBJECT_SAMPLE_FACTORS - WL_RPLC_POS_WT_2 Genotype:Wild-type | Sample source:Whole Larvae Chromatography=RPLC; Polarity=Positive; RAW_FILE_NAME(Raw file name)=WL_RPLC_POS_WT_2.mzML SUBJECT_SAMPLE_FACTORS - WL_RPLC_POS_WT_3 Genotype:Wild-type | Sample source:Whole Larvae Chromatography=RPLC; Polarity=Positive; RAW_FILE_NAME(Raw file name)=WL_RPLC_POS_WT_3.mzML SUBJECT_SAMPLE_FACTORS - WL_RPLC_POS_WT_4 Genotype:Wild-type | Sample source:Whole Larvae Chromatography=RPLC; Polarity=Positive; RAW_FILE_NAME(Raw file name)=WL_RPLC_POS_WT_4.mzML SUBJECT_SAMPLE_FACTORS - WL_RPLC_POS_WT_5 Genotype:Wild-type | Sample source:Whole Larvae Chromatography=RPLC; Polarity=Positive; RAW_FILE_NAME(Raw file name)=WL_RPLC_POS_WT_5.mzML SUBJECT_SAMPLE_FACTORS - CNS_RPLC_POS_ifc_1 Genotype:Knockout | Sample source:CNS Chromatography=RPLC; Polarity=Positive; RAW_FILE_NAME(Raw file name)=CNS_RPLC_POS_ifc_1.mzML SUBJECT_SAMPLE_FACTORS - CNS_RPLC_POS_ifc_2 Genotype:Knockout | Sample source:CNS Chromatography=RPLC; Polarity=Positive; RAW_FILE_NAME(Raw file name)=CNS_RPLC_POS_ifc_2.mzML SUBJECT_SAMPLE_FACTORS - CNS_RPLC_POS_ifc_3 Genotype:Knockout | Sample source:CNS Chromatography=RPLC; Polarity=Positive; RAW_FILE_NAME(Raw file name)=CNS_RPLC_POS_ifc_3.mzML SUBJECT_SAMPLE_FACTORS - CNS_RPLC_POS_ifc_4 Genotype:Knockout | Sample source:CNS Chromatography=RPLC; Polarity=Positive; RAW_FILE_NAME(Raw file name)=CNS_RPLC_POS_ifc_4.mzML SUBJECT_SAMPLE_FACTORS - CNS_RPLC_POS_ifc_5 Genotype:Knockout | Sample source:CNS Chromatography=RPLC; Polarity=Positive; RAW_FILE_NAME(Raw file name)=CNS_RPLC_POS_ifc_5.mzML SUBJECT_SAMPLE_FACTORS - CNS_RPLC_POS_WT_1 Genotype:Wild-type | Sample source:CNS Chromatography=RPLC; Polarity=Positive; RAW_FILE_NAME(Raw file name)=CNS_RPLC_POS_WT_1.mzML SUBJECT_SAMPLE_FACTORS - CNS_RPLC_POS_WT_2 Genotype:Wild-type | Sample source:CNS Chromatography=RPLC; Polarity=Positive; RAW_FILE_NAME(Raw file name)=CNS_RPLC_POS_WT_2.mzML SUBJECT_SAMPLE_FACTORS - CNS_RPLC_POS_WT_3 Genotype:Wild-type | Sample source:CNS Chromatography=RPLC; Polarity=Positive; RAW_FILE_NAME(Raw file name)=CNS_RPLC_POS_WT_3.mzML SUBJECT_SAMPLE_FACTORS - CNS_RPLC_POS_WT_4 Genotype:Wild-type | Sample source:CNS Chromatography=RPLC; Polarity=Positive; RAW_FILE_NAME(Raw file name)=CNS_RPLC_POS_WT_4.mzML SUBJECT_SAMPLE_FACTORS - CNS_RPLC_POS_WT_5 Genotype:Wild-type | Sample source:CNS Chromatography=RPLC; Polarity=Positive; RAW_FILE_NAME(Raw file name)=CNS_RPLC_POS_WT_5.mzML SUBJECT_SAMPLE_FACTORS - WL_HILIC_NEG_ifc_1 Genotype:Knockout | Sample source:Whole Larvae Chromatography=HILIC; Polarity=Negative; RAW_FILE_NAME(Raw file name)=WL_HILIC_NEG_ifc_1.mzML SUBJECT_SAMPLE_FACTORS - WL_HILIC_NEG_ifc_2 Genotype:Knockout | Sample source:Whole Larvae Chromatography=HILIC; Polarity=Negative; RAW_FILE_NAME(Raw file name)=WL_HILIC_NEG_ifc_2.mzML SUBJECT_SAMPLE_FACTORS - WL_HILIC_NEG_ifc_3 Genotype:Knockout | Sample source:Whole Larvae Chromatography=HILIC; Polarity=Negative; RAW_FILE_NAME(Raw file name)=WL_HILIC_NEG_ifc_3.mzML SUBJECT_SAMPLE_FACTORS - WL_HILIC_NEG_ifc_4 Genotype:Knockout | Sample source:Whole Larvae Chromatography=HILIC; Polarity=Negative; RAW_FILE_NAME(Raw file name)=WL_HILIC_NEG_ifc_4.mzML SUBJECT_SAMPLE_FACTORS - WL_HILIC_NEG_ifc_5 Genotype:Knockout | Sample source:Whole Larvae Chromatography=HILIC; Polarity=Negative; RAW_FILE_NAME(Raw file name)=WL_HILIC_NEG_ifc_5.mzML SUBJECT_SAMPLE_FACTORS - WL_HILIC_NEG_WT_1 Genotype:Wild-type | Sample source:Whole Larvae Chromatography=HILIC; Polarity=Negative; RAW_FILE_NAME(Raw file name)=WL_HILIC_NEG_WT_1.mzML SUBJECT_SAMPLE_FACTORS - WL_HILIC_NEG_WT_2 Genotype:Wild-type | Sample source:Whole Larvae Chromatography=HILIC; Polarity=Negative; RAW_FILE_NAME(Raw file name)=WL_HILIC_NEG_WT_2.mzML SUBJECT_SAMPLE_FACTORS - WL_HILIC_NEG_WT_3 Genotype:Wild-type | Sample source:Whole Larvae Chromatography=HILIC; Polarity=Negative; RAW_FILE_NAME(Raw file name)=WL_HILIC_NEG_WT_3.mzML SUBJECT_SAMPLE_FACTORS - WL_HILIC_NEG_WT_4 Genotype:Wild-type | Sample source:Whole Larvae Chromatography=HILIC; Polarity=Negative; RAW_FILE_NAME(Raw file name)=WL_HILIC_NEG_WT_4.mzML SUBJECT_SAMPLE_FACTORS - WL_HILIC_NEG_WT_5 Genotype:Wild-type | Sample source:Whole Larvae Chromatography=HILIC; Polarity=Negative; RAW_FILE_NAME(Raw file name)=WL_HILIC_NEG_WT_5.mzML SUBJECT_SAMPLE_FACTORS - CNS_HILIC_NEG_ifc_1 Genotype:Knockout | Sample source:CNS Chromatography=HILIC; Polarity=Negative; RAW_FILE_NAME(Raw file name)=CNS_HILIC_NEG_ifc_1.mzML SUBJECT_SAMPLE_FACTORS - CNS_HILIC_NEG_ifc_2 Genotype:Knockout | Sample source:CNS Chromatography=HILIC; Polarity=Negative; RAW_FILE_NAME(Raw file name)=CNS_HILIC_NEG_ifc_2.mzML SUBJECT_SAMPLE_FACTORS - CNS_HILIC_NEG_ifc_3 Genotype:Knockout | Sample source:CNS Chromatography=HILIC; Polarity=Negative; RAW_FILE_NAME(Raw file name)=CNS_HILIC_NEG_ifc_3.mzML SUBJECT_SAMPLE_FACTORS - CNS_HILIC_NEG_ifc_4 Genotype:Knockout | Sample source:CNS Chromatography=HILIC; Polarity=Negative; RAW_FILE_NAME(Raw file name)=CNS_HILIC_NEG_ifc_4.mzML SUBJECT_SAMPLE_FACTORS - CNS_HILIC_NEG_ifc_5 Genotype:Knockout | Sample source:CNS Chromatography=HILIC; Polarity=Negative; RAW_FILE_NAME(Raw file name)=CNS_HILIC_NEG_ifc_5.mzML SUBJECT_SAMPLE_FACTORS - CNS_HILIC_NEG_WT_1 Genotype:Wild-type | Sample source:CNS Chromatography=HILIC; Polarity=Negative; RAW_FILE_NAME(Raw file name)=CNS_HILIC_NEG_WT_1.mzML SUBJECT_SAMPLE_FACTORS - CNS_HILIC_NEG_WT_2 Genotype:Wild-type | Sample source:CNS Chromatography=HILIC; Polarity=Negative; RAW_FILE_NAME(Raw file name)=CNS_HILIC_NEG_WT_2.mzML SUBJECT_SAMPLE_FACTORS - CNS_HILIC_NEG_WT_3 Genotype:Wild-type | Sample source:CNS Chromatography=HILIC; Polarity=Negative; RAW_FILE_NAME(Raw file name)=CNS_HILIC_NEG_WT_3.mzML SUBJECT_SAMPLE_FACTORS - CNS_HILIC_NEG_WT_4 Genotype:Wild-type | Sample source:CNS Chromatography=HILIC; Polarity=Negative; RAW_FILE_NAME(Raw file name)=CNS_HILIC_NEG_WT_4.mzML SUBJECT_SAMPLE_FACTORS - CNS_HILIC_NEG_WT_5 Genotype:Wild-type | Sample source:CNS Chromatography=HILIC; Polarity=Negative; RAW_FILE_NAME(Raw file name)=CNS_HILIC_NEG_WT_5.mzML #COLLECTION CO:COLLECTION_SUMMARY Whole larvae and dissected CNS CO:SAMPLE_TYPE Larvae #TREATMENT TR:TREATMENT_SUMMARY Untargeted lipidomics analysis was conducted on whole larva and dissected CNS of TR:TREATMENT_SUMMARY wild type and ifc-/- mutants at the late-third instar stage. Five replicates TR:TREATMENT_SUMMARY were prepared for each set of experiments. For whole larvae, at least 15 larvae TR:TREATMENT_SUMMARY of each genotype were used for each replicate. For the dissected CNS, at least TR:TREATMENT_SUMMARY 50 wild type and 60 ifc-/- CNS were used per replicate. Immediately following TR:TREATMENT_SUMMARY collection or dissection, larvae and the dissected CNS were flash frozen in TR:TREATMENT_SUMMARY liquid nitrogen and placed at -80°C. #SAMPLEPREP SP:SAMPLEPREP_SUMMARY Whole larvae and CNS samples were extracted utilizing a tissue homogenizer, SP:SAMPLEPREP_SUMMARY employing a solvent mixture of acetonitrile, methanol, and water in a 2:2:1 SP:SAMPLEPREP_SUMMARY ratio, using 40 µL per mg of wet weight. The samples were then stored at -20°C SP:SAMPLEPREP_SUMMARY overnight. Following centrifugation at 14,000 x g for 10 minutes at 4°C, the SP:SAMPLEPREP_SUMMARY supernatant was transferred into an LC/MS vial and subsequently stored at -80°C SP:SAMPLEPREP_SUMMARY until LC/MS analysis. #CHROMATOGRAPHY CH:CHROMATOGRAPHY_TYPE HILIC CH:INSTRUMENT_NAME Thermo Vanquish Flex UHPLC Systems CH:COLUMN_NAME HILICON iHILIC-(P) Classic (100 x 2.1mm,5um) CH:SOLVENT_A water/acetonitrile (95/5); 20 mM ammonium bicarbonate; 0.1% ammonium hydroxide; CH:SOLVENT_A 2.5 μM medronic acid CH:SOLVENT_B acetonitrile/water (95/5) CH:FLOW_GRADIENT 0–1 min: 90% B, 1–12 min: 90-35% B, 12–12.5 min: 35-25% B, 12.5–14.5 CH:FLOW_GRADIENT min: 25% B CH:FLOW_RATE 250 uL/min CH:COLUMN_TEMPERATURE 45 #ANALYSIS AN:ANALYSIS_TYPE MS #MS MS:INSTRUMENT_NAME Thermo Orbitrap ID-X tribrid MS:INSTRUMENT_TYPE Orbitrap MS:MS_TYPE ESI MS:ION_MODE NEGATIVE MS:MS_COMMENTS Data were collected with the following MS source settings: spray voltage, -2.8 MS:MS_COMMENTS kV; sheath gas, 50; auxiliary gas, 10; sweep gas, 1; ion transfer tube MS:MS_COMMENTS temperature, 300°C; vaporizer temperature, 200°C; mass range, 67 – 1000 Da; MS:MS_COMMENTS resolution, 120,000; maximum injection time, 200 ms; isolation window, 1.6 Da. MS:MS_COMMENTS XCMS and Skyline software were used for data processing #MS_METABOLITE_DATA MS_METABOLITE_DATA:UNITS peak area MS_METABOLITE_DATA_START Samples WL_HILIC_NEG_ifc_1 WL_HILIC_NEG_ifc_2 WL_HILIC_NEG_ifc_3 WL_HILIC_NEG_ifc_4 WL_HILIC_NEG_ifc_5 WL_HILIC_NEG_WT_1 WL_HILIC_NEG_WT_2 WL_HILIC_NEG_WT_3 WL_HILIC_NEG_WT_4 WL_HILIC_NEG_WT_5 Factors Genotype:Knockout | Sample source:Whole Larvae Genotype:Knockout | Sample source:Whole Larvae Genotype:Knockout | Sample source:Whole Larvae Genotype:Knockout | Sample source:Whole Larvae Genotype:Knockout | Sample source:Whole Larvae Genotype:Wild-type | Sample source:Whole Larvae Genotype:Wild-type | Sample source:Whole Larvae Genotype:Wild-type | Sample source:Whole Larvae Genotype:Wild-type | Sample source:Whole Larvae Genotype:Wild-type | Sample source:Whole Larvae Glycine 15490151 14902291 14800931 11965879 13191878 10866370 10418561 12671627 10180559 9230180 Pyruvate 75795600 92614416 92902968 92419440 120535416 90136560 108469640 90679392 135537168 121134232 Alanine 151069088 148102400 155855520 144975808 166736464 152576528 157817600 147928496 195312144 176193888 Lactate 1558859264 1466138112 1558577152 1549909248 1635084032 1820667136 1885712000 1697844480 1828337280 1990820992 Serine 24373358 22281050 23501476 18258446 25389750 19565970 18413790 22111544 25037630 20182396 Phosphoenolpyruvate 186955 202809 211829 204288 388909 145273 163675 152855 215471 207921 DHAP 642653 639217 660695 424566 938418 485012 389331 452666 624500 494311 Glycerol 3-phosphate 311892320 317801504 315135968 274590976 340962592 185773664 167372384 164554528 228222608 199037760 2/3-PG 4972382 4731640 4738588 3139438 6324483 2875048 2550338 1940046 2851370 3197174 Fumarate 38787704 41869352 40791252 42520500 45543864 66441924 67648840 62906248 73228032 59112084 Succinate 299368160 309094208 307720864 309724032 331684384 334267968 343217184 270159104 370888512 362578016 Asparagine 63954124 65484924 62377288 49054232 67578216 61223344 58213672 55062856 68886160 59043812 Aspartate 24624244 23184952 22436616 17396618 18357138 18938438 17361946 17135504 15732642 12866094 Malate 614096192 658213888 634738880 647661184 682715136 973887872 977918400 931896960 1052045504 868698816 Threonine 120548552 93381968 103495392 66899680 94681624 49307012 45112060 47531656 41407672 40616448 Proline 591293568 584489472 587373888 554899776 606629888 496029760 477944640 448740640 576244224 540356160 Valine 170729680 149108704 156488704 139301552 138327344 87129456 90074824 92874280 103075208 87895176 Leucine 175854672 114589360 121745504 363600448 320666080 129076888 125734384 261975360 136002144 131711864 Adenine 16476203 14823735 15921752 14602692 11355252 52206084 51791212 38700560 40751160 33736368 a-KG 39250432 46009904 47715100 55099768 62986104 33576372 41105808 40766336 56577848 47686096 Glutamine 371569088 382721440 370045984 282810496 419062752 279796768 267894592 250782112 303843200 254150304 Glutamate 569432192 530808672 552301888 480074464 606459968 364468544 344294304 355488064 422397184 365526272 Methionine 36509300 29424990 33960688 33753696 27690956 30203250 30539880 32314192 35910760 31428256 Ribose-5-phosphate 544287 660980 692213 680299 809439 651940 433752 599743 472537 588317 Glucose-6-phosphate 213341392 180088080 203220656 185656688 201382464 189699648 175150640 168408528 212247088 208724240 Lysine 19748784 20348644 19676354 13567196 19884514 13515924 13290924 11615638 14879620 13255645 Aconitate 13232557 17344150 15644301 17911366 20649208 12378993 12427496 13216969 24081614 17353038 Arginine 110815600 96713680 100462600 81608136 103906992 93018256 87759976 76523840 101934720 91601488 Glucose 68964232 73849520 73270584 67098236 69160552 60371764 57895036 61638404 53402680 53816924 Citrate 345995904 382549728 360204320 372447936 433066560 401645408 392863232 366632544 450848672 401976544 Fructose-bisphosphate 592595 428944 436573 250852 495262 398338 310758 230704 410509 369046 6-Phosphogluconic acid 281328 225958 249168 178631 218974 271632 173877 153110 259241 232455 Phenylalanine 114550336 112490432 112282976 109503672 107717680 168050640 153785696 152609568 147564624 146416416 Tyrosine 898784768 824117632 949566912 1065398464 1055201600 2455355392 2170336768 2149325312 2048419200 1923725184 UMP 3111713 3082369 3058718 2917545 3009174 6550380 5130187 6227417 7423256 5030959 Glutathione 151859776 151299760 146128784 125279816 162622192 137747888 141461472 126013512 159667696 116162824 AMP 213472560 196293328 211318608 209088720 213423424 303616224 286119936 261324480 327575456 313469216 ADP 24661628 22039076 22540032 18761936 21390488 19139698 14371489 12858274 17487246 17052918 ATP 3312118 2736030 2819218 1796568 3487297 1354590 944504 834356 1336684 1039939 Uridine 5'-diphosphoglucuronic acid 315170 285837 292697 228966 282412 324975 331861 262293 309280 246689 UDP-N-acetylglucosamine 13317971 12287296 14007477 10688915 21590266 18846534 18802250 16431084 27072574 22884902 Glutathione, oxidized 590461 515991 517186 395735 419984 448260 370672 293557 346983 264156 NAD 7269626 6463837 6952030 7328879 7916512 10796759 10596752 8668649 11834088 9614821 NADH 2107868 1742286 2029091 1307620 2534949 2700381 1959562 1451450 2545633 1914703 NADPH 523399 502638 511903 341241 486034 655801 545988 530349 589277 478769 FAD 2352128 2106733 2413400 1817261 2756483 2731490 2649915 2411009 2810640 2412997 Xylulose-5-phosphate 3292868 2593090 2963969 2022793 3447758 4287582 3969395 3199786 4239224 4601813 Ribulose-5-phosphate 754718 249646 778285 569297 381354 724126 326018 575137 271203 274405 Gluconate 73549432 73984792 66154168 42996284 59011864 33018062 31381026 27073778 34328528 37502072 Sedoheptulose 7-phosphate 18726060 17885316 18860534 18687560 17764796 25495894 23961852 21577656 32810602 27946676 Isoleucine 166440800 278610592 307868608 221625600 211656496 102145032 148952736 170383984 190706704 127284000 MS_METABOLITE_DATA_END #METABOLITES METABOLITES_START metabolite_name Glycine Pyruvate Alanine Lactate Serine Phosphoenolpyruvate DHAP Glycerol 3-phosphate 2/3-PG Fumarate Succinate Asparagine Aspartate Malate Threonine Proline Valine Leucine Adenine a-KG Glutamine Glutamate Methionine Ribose-5-phosphate Glucose-6-phosphate Lysine Aconitate Arginine Glucose Citrate Fructose-bisphosphate 6-Phosphogluconic acid Phenylalanine Tyrosine UMP Glutathione AMP ADP ATP Uridine 5'-diphosphoglucuronic acid UDP-N-acetylglucosamine Glutathione, oxidized NAD NADH NADPH FAD Xylulose-5-phosphate Ribulose-5-phosphate Gluconate Sedoheptulose 7-phosphate Isoleucine METABOLITES_END #END