#METABOLOMICS WORKBENCH QiushiSun_20221128_085601 DATATRACK_ID:3602 STUDY_ID:ST002361 ANALYSIS_ID:AN003856 VERSION 1 CREATED_ON 06-06-2023 #PROJECT PR:PROJECT_TITLE UCP2-dependent redox-sensing in POMC neurons regulates feeding PR:PROJECT_SUMMARY Paradoxically, glucose, the primary driver of satiety, activates a small PR:PROJECT_SUMMARY population of anorexigenic POMC neurons. Here we show that lactate levels in the PR:PROJECT_SUMMARY circulation and in the cerebrospinal fluid are elevated in fed state and PR:PROJECT_SUMMARY addition of lactate to glucose activates the majority of POMC neurons while PR:PROJECT_SUMMARY increasing cytosolic NADH generation, mitochondrial respiration and PR:PROJECT_SUMMARY extracellular pyruvate levels. Inhibition of lactate dehydrogenases diminishes PR:PROJECT_SUMMARY mitochondrial respiration, NADH production, and POMC neuronal activity. However, PR:PROJECT_SUMMARY inhibition of the mitochondrial pyruvate carrier has no effect. POMC-specific PR:PROJECT_SUMMARY downregulation of Ucp2 (Ucp2PomcKO), a molecule regulated by fatty acid PR:PROJECT_SUMMARY metabolism and shown to play a role as transporter in the malate-aspartate PR:PROJECT_SUMMARY shuttle, abolishes lactate- and glucose-sensing of POMC neurons. Ucp2PomcKO mice PR:PROJECT_SUMMARY have impaired glucose metabolism and are prone to obesity on a high fat diet. PR:PROJECT_SUMMARY Altogether, our data show that lactate through redox signaling and blocking PR:PROJECT_SUMMARY mitochondrial glucose utilization activates POMC neurons to regulate feeding and PR:PROJECT_SUMMARY glucose metabolism. PR:INSTITUTE Columbia University PR:LAST_NAME Diano PR:FIRST_NAME Sabrina PR:ADDRESS 1150 St. Nicholas Avenue Russ Berrie Medical Science Pavilion Rm 405 New York, PR:ADDRESS NY, 10032 PR:EMAIL sd3449@cumc.columbia.edu PR:PHONE 212 8514554 PR:DOI http://dx.doi.org/10.21228/M8R12T #STUDY ST:STUDY_TITLE UCP2-dependent redox-sensing in POMC neurons regulates feeding ST:STUDY_SUMMARY Paradoxically, glucose, the primary driver of satiety, activates a small ST:STUDY_SUMMARY population of anorexigenic POMC neurons. Here we show that lactate levels in the ST:STUDY_SUMMARY circulation and in the cerebrospinal fluid are elevated in fed state and ST:STUDY_SUMMARY addition of lactate to glucose activates the majority of POMC neurons while ST:STUDY_SUMMARY increasing cytosolic NADH generation, mitochondrial respiration and ST:STUDY_SUMMARY extracellular pyruvate levels. Inhibition of lactate dehydrogenases diminishes ST:STUDY_SUMMARY mitochondrial respiration, NADH production, and POMC neuronal activity. However, ST:STUDY_SUMMARY inhibition of the mitochondrial pyruvate carrier has no effect. POMC-specific ST:STUDY_SUMMARY downregulation of Ucp2 (Ucp2PomcKO), a molecule regulated by fatty acid ST:STUDY_SUMMARY metabolism and shown to play a role as transporter in the malate-aspartate ST:STUDY_SUMMARY shuttle, abolishes lactate- and glucose-sensing of POMC neurons. Ucp2PomcKO mice ST:STUDY_SUMMARY have impaired glucose metabolism and are prone to obesity on a high fat diet. ST:STUDY_SUMMARY Altogether, our data show that lactate through redox signaling and blocking ST:STUDY_SUMMARY mitochondrial glucose utilization activates POMC neurons to regulate feeding and ST:STUDY_SUMMARY glucose metabolism. ST:INSTITUTE Columbia University ST:LAST_NAME Diano ST:FIRST_NAME Sabrina ST:ADDRESS 1150 St. Nicholas Avenue Russ Berrie Medical Science Pavilion Rm 405 New York, ST:ADDRESS NY, 10032 ST:EMAIL sd3449@cumc.columbia.edu ST:PHONE 212 8514554 ST:SUBMIT_DATE 2022-11-28 #SUBJECT SU:SUBJECT_TYPE Cultured cells SU:SUBJECT_SPECIES Mus musculus SU:TAXONOMY_ID 10090 #SUBJECT_SAMPLE_FACTORS: SUBJECT(optional)[tab]SAMPLE[tab]FACTORS(NAME:VALUE pairs separated by |)[tab]Additional sample data SUBJECT_SAMPLE_FACTORS - Ucp2PomcKO_1_HC_Neg Genotype:UCP2 POMC-knockout Instrument run modes=Hypercarb column negative MS mode (HC_Neg); RAW_FILE_NAME=run15-16Sep22_KO_GlcNA_LacNA_1_HC_Neg.mzML SUBJECT_SAMPLE_FACTORS - Ucp2PomcKO_1_RP_Pos Genotype:UCP2 POMC-knockout Instrument run modes=Reverse phase column positive MS mode (RP_Pos); RAW_FILE_NAME=run15-15Sep22_KO_GlcNA_LacNA_1_RP_Pos.mzML SUBJECT_SAMPLE_FACTORS - Ucp2PomcKO_2_HC_Neg Genotype:UCP2 POMC-knockout Instrument run modes=Hypercarb column negative MS mode (HC_Neg); RAW_FILE_NAME=run16-16Sep22_KO_GlcNA_LacNA_2_HC_Neg.mzML SUBJECT_SAMPLE_FACTORS - Ucp2PomcKO_2_RP_Pos Genotype:UCP2 POMC-knockout Instrument run modes=Reverse phase column positive MS mode (RP_Pos); RAW_FILE_NAME=run16-15Sep22_KO_GlcNA_LacNA_2_RP_Pos.mzML SUBJECT_SAMPLE_FACTORS - Ctrl_1_HC_Neg Genotype:wild type Instrument run modes=Hypercarb column negative MS mode (HC_Neg); RAW_FILE_NAME=run6-16Sep22_Ctrl_GlcNA_LacNA_1_HC_Neg.mzML SUBJECT_SAMPLE_FACTORS - Ctrl_1_RP_Pos Genotype:wild type Instrument run modes=Reverse phase column positive MS mode (RP_Pos); RAW_FILE_NAME=run6-15Sep22_Ctrl_GlcNA_LacNA_1_RP_Pos.mzML SUBJECT_SAMPLE_FACTORS - Ctrl_2_HC_Neg Genotype:wild type Instrument run modes=Hypercarb column negative MS mode (HC_Neg); RAW_FILE_NAME=run25-16Sep22_Ctrl_GlcNA_LacNA_2_HC_Neg_rerun.mzML SUBJECT_SAMPLE_FACTORS - Ctrl_2_RP_Pos Genotype:wild type Instrument run modes=Reverse phase column positive MS mode (RP_Pos); RAW_FILE_NAME=run7-15Sep22_Ctrl_GlcNA_LacNA_2_RP_Pos.mzML #COLLECTION CO:COLLECTION_SUMMARY Hypothalamic primary neuronal cell culture Eight to ten neonatal (0 days old) CO:COLLECTION_SUMMARY pubs from either control or Ucp2PomcKO mice derived from homozygous Cre-positive CO:COLLECTION_SUMMARY parents were used for hypothalamic primary neuronal cell culture. In brief, we CO:COLLECTION_SUMMARY carefully removed the brain's hypothalamus and placed it onto a small culture CO:COLLECTION_SUMMARY dish containing a small volume of Hibernate-A Medium (Cat# A1247501, Thermo CO:COLLECTION_SUMMARY Fisher Scientific). After digestion, the tissues dissociated to single cells CO:COLLECTION_SUMMARY with 6 mL of Hibernate-A Medium containing 2.5 % of Trypsin-EDTA for 15 minutes CO:COLLECTION_SUMMARY at 37℃. Suspended cells were filtered (40 μm) and centrifuged for 5 min at CO:COLLECTION_SUMMARY 1000 rpm. The pellet was re-suspended and plated on XF96 cell culture CO:COLLECTION_SUMMARY microplates (Cat# 101085-004, Agilent Technologies) coated with poly-D-lysine CO:COLLECTION_SUMMARY (Cat# P6407, Sigma-Aldrich) at a density of 0.2 x104 cells per well. Cells were CO:COLLECTION_SUMMARY cultured in Neurobasal-A medium (Cat# 10888022, Thermo Fisher Scientific) CO:COLLECTION_SUMMARY supplemented with 1 % penicillin-streptomycin, 2 % B-27 Supplement (Cat# CO:COLLECTION_SUMMARY 17504044, Thermo Fisher Scientific), and GlutaMAX-I (Cat# 35050061, Thermo CO:COLLECTION_SUMMARY Fisher Scientific), CultureOne supplement (Cat# A3320201, Thermo Fisher CO:COLLECTION_SUMMARY Scientific). For control culture, we used either Ucp2fl/fl; Pomc-CreERT2; CO:COLLECTION_SUMMARY tdTomato mice which neuronal cultures were treated with vehicle (0.03 % ethanol CO:COLLECTION_SUMMARY diluted in medium) or Ucp2+/+; Pomc-CreERT2; tdTomato mice which neuronal CO:COLLECTION_SUMMARY cultures were treated with 2 μM 4-hydroxytamoxifen (Cat# H7904, Sigma-Aldrich). CO:COLLECTION_SUMMARY After 10 days in culture, primary neuronal cells isolated from control and CO:COLLECTION_SUMMARY Ucp2PomcKO mice were treated with 2 μM 4-hydroxytamoxifen for expression of a CO:COLLECTION_SUMMARY CreER recombinase. Primary neuronal cells were used for the measurement of CO:COLLECTION_SUMMARY mitochondria oxidation two days later. CO:SAMPLE_TYPE Neurons #TREATMENT TR:TREATMENT_SUMMARY We used the inducible Cre/loxP technology to generate mice in which UCP2 was TR:TREATMENT_SUMMARY selectively ablated in POMC neurons (Ucp2PomcKO mice). First, mice expressing a TR:TREATMENT_SUMMARY tamoxifen-inducible Cre recombinase (CreERT2) in cells expressing POMC TR:TREATMENT_SUMMARY (Pomc-CreERT2) were crossed with Rosa26-lox-stop-lox-tdTomato (Ai14; TR:TREATMENT_SUMMARY cre-recombinase-dependent expression) mice (Ai14 reporter mice; stock #007914; TR:TREATMENT_SUMMARY The Jackson Laboratory) to label POMC-expressing cells. Pomc-CreERT2; TR:TREATMENT_SUMMARY Rosa26-lox-stop-lox-tdTomato (Pomc-CreERT2; tdTomato) mice have POMC-expressing TR:TREATMENT_SUMMARY cells with the expression of tdTomato by tamoxifen administration. No TR:TREATMENT_SUMMARY observation of POMC-tdTomato expression was found in the absence of tamoxifen TR:TREATMENT_SUMMARY administration, indicating that recombination was strictly dependent upon TR:TREATMENT_SUMMARY tamoxifen-induced Cre recombinase activation. The mice with Pomc-CreERT2; TR:TREATMENT_SUMMARY tdTomato were then crossed with mice harboring conditional alleles Ucp2 floxed TR:TREATMENT_SUMMARY (Ucp2fl/fl; B6;129S-Ucp2tm2.1Lowl/J, Stock# 022394; The Jackson Laboratory) to TR:TREATMENT_SUMMARY generated mice with inducible deletion of Ucp2 specifically in POMC neurons TR:TREATMENT_SUMMARY (Ucp2PomcKO mice). All animal experiments were conducted with the mice TR:TREATMENT_SUMMARY expressing POMC-CreERT2. As control groups, Ucp2fl/fl; Pomc-CreERT2 mice were TR:TREATMENT_SUMMARY injected with tamoxifen, and Ucp2fl/fl; Pomc-CreERT2; tdTomato were mice TR:TREATMENT_SUMMARY injected with corn oil (vehicle), and Ucp2fl/fl; Pomc-CreERT2; tdTomato mice TR:TREATMENT_SUMMARY were injected intraperitoneally (IP) with tamoxifen (0.1 mg/g BW for 5 TR:TREATMENT_SUMMARY consecutive days) starting at 5 weeks of age to induce mature-onset deletion of TR:TREATMENT_SUMMARY Ucp2 in POMC neurons of Ucp2PomcKO mice. However, because there were no TR:TREATMENT_SUMMARY differences between these two control groups, most of the experiments were TR:TREATMENT_SUMMARY performed using Ucp2+/+; Pomc-CreERT2; tdTomato mice injected with tamoxifen (to TR:TREATMENT_SUMMARY label POMC neurons with tdTomato expression) as a control group, unless TR:TREATMENT_SUMMARY otherwise stated. Body composition was measured in vivo by MRI (EchoMRI; Echo TR:TREATMENT_SUMMARY Medical Systems, Houston, TX) monthly at 10:00 AM. We performed transcriptomic TR:TREATMENT_SUMMARY profiling by using a ribosomal tagging strategy to analyze POMC neuron-specific TR:TREATMENT_SUMMARY mRNA expression in vivo. We crossed Pomc-CreERT2 mice30 with Rpl22 floxed TR:TREATMENT_SUMMARY (RiboTag, Stock# 029977, The Jackson Laboratories) mice to generate TR:TREATMENT_SUMMARY Pomc-CreERT2; RiboTag mice, expressing a hemagglutinin A (HA)-tagged ribosomal TR:TREATMENT_SUMMARY protein in the POMC neurons upon tamoxifen injection. #SAMPLEPREP SP:SAMPLEPREP_SUMMARY Cells cultured in 6-well plates were quickly washed once with ice cold 5 mM SP:SAMPLEPREP_SUMMARY HEPES, then immediately quenched in 150ul/well ice cold quench buffer (20% SP:SAMPLEPREP_SUMMARY methanol, 3 mM sodium fluoride, 0.1% formic acid, 1mM phenylalanine and 100uM SP:SAMPLEPREP_SUMMARY EDTA). Each well was harvested using a cell lifter and immediately transferred SP:SAMPLEPREP_SUMMARY to a pre-chilled 96-well plate on dry ice. Once completely frozen, cell lysates SP:SAMPLEPREP_SUMMARY were lyophilized and stored in the -80oC freezer until the sample run. SP:SAMPLEPREP_SUMMARY Lyophilized samples were prepared by resuspending in 50 µL water with SP:SAMPLEPREP_SUMMARY D4-taurine (50 µM) as an internal standard and 5 µL of the supernatant was SP:SAMPLEPREP_SUMMARY injected for each analysis mode using high performance liquid chromatography SP:SAMPLEPREP_SUMMARY (HPLC) into the 6600 Triple TOF LC-MS/MS mass spectrometer (Sciex). #CHROMATOGRAPHY CH:CHROMATOGRAPHY_SUMMARY Hypercarb column with negative MS mode CH:INSTRUMENT_NAME SCIEX ExionLC CH:COLUMN_NAME Thermo Scientific Hypercarb (100 x 4.6 mm,3um) CH:COLUMN_TEMPERATURE 50 CH:FLOW_RATE 1ml/min CH:SOLVENT_A 100% water; 15 mM ammonium formate; 0.03% acetyl acetone; 0.1% formic acid CH:SOLVENT_B 60% acetonitrile/35% isopropanol; 0.1% formic acid; 15 mM ammonium formate CH:ANALYTICAL_TIME 18min CH:WASHING_BUFFER 50%Methanol, 50% water CH:CHROMATOGRAPHY_TYPE Other #ANALYSIS AN:ANALYSIS_TYPE MS AN:ANALYSIS_PROTOCOL_FILE MS_Analysis_Protocol.pdf #MS MS:INSTRUMENT_NAME ABI Sciex 6600 TripleTOF MS:INSTRUMENT_TYPE Triple TOF MS:MS_TYPE ESI MS:MS_COMMENTS Data were collected using an information-dependent analysis (IDA) workflow MS:MS_COMMENTS consisting of a TOF MS scan (200 msec) and a high-resolution IDA experiment (70 MS:MS_COMMENTS msec each) monitoring 10 candidate ions per cycle. Former target ions were MS:MS_COMMENTS excluded after 2 occurrences for 5 seconds and dynamic background subtraction MS:MS_COMMENTS was employed. The mass range for both TOF MS and IDA MS/MS scans was 70-1000. MS:MS_COMMENTS Ion spray voltage = -4500V, ion source gas 1 (GS1) = 50 psi, ion source gas 2 MS:MS_COMMENTS (GS2) = 50 psi, curtain gas (CUR) = 30 psi, temperature (TEM) = 500 MS:MS_COMMENTS oC.declustering potential (DP) = 35 V, collision energy (CE) = 30 V, collision MS:MS_COMMENTS energy spread (CES) = 20 V. El-MAVEN software (Elucidata.io) was used for peak MS:MS_COMMENTS picking and curation from house built targeted and untargeted libraries. MS:MS_COMMENTS Targeted libraries were developed using a commercial standard kit of ~600 MS:MS_COMMENTS metabolites (IROA Technologies) in each mode, yielding reference data (molecular MS:MS_COMMENTS ions and retention times) with wide coverage of the endogenous metabolome. MS:MS_COMMENTS Untargeted libraries contained ~2,700 metabolites drawn from the KEGG database MS:MS_COMMENTS where the top 5 candidates with the highest intensity throughout a sample run MS:MS_COMMENTS for each metabolite were automatically curated. MS:ION_MODE NEGATIVE #MS_METABOLITE_DATA MS_METABOLITE_DATA:UNITS peak area MS_METABOLITE_DATA_START Samples Ucp2PomcKO_1_HC_Neg Ucp2PomcKO_2_HC_Neg Ctrl_1_HC_Neg Ctrl_2_HC_Neg Factors Genotype:UCP2 POMC-knockout Genotype:UCP2 POMC-knockout Genotype:wild type Genotype:wild type 10-HYDROXYDECANOATE 2189.0000 2781.0000 2410.0000 2795.0000 (2-AMINOETHYL)PHOSPHONATE 5671.0000 8785.0000 4349.0000 3760.0000 2-HYDROXYBUTYRATE 519.0000 680.0000 893.6000 648.2300 3-(2-HYDROXYPHENYL)PROPANOATE 435197.5000 655237.4400 674339.3100 401900.8100 3_4 DIHYDROXYMANDELATE 58084.0000 70620.4500 76693.0000 60630.4200 3_4-DIHYDROXYPHENYLGLYCOL 6559934.5000 8762054.0000 8526764.0000 6471353.0000 3-HYDROXYMETHYLGLUTARATE 86030.2100 151044.6400 165546.9100 100581.2700 3-METHOXY-4-HYDROXYMANDELATE 394651.8800 602151.2500 272594.0000 472837.9400 3-SULFINOALANINE 3606.0000 6168.0000 2760.0000 1590.0000 4-HYDROXYBENZALDEHYDE 11990.7400 9907.3900 10568.4400 9988.0500 5-AMINOLEVULINATE 363.0000 563.0000 520.0000 554.0000 5-AMINOPENTANOATE 22908.0000 24784.0000 13146.0000 16534.0000 6-CARBOXYHEXANOATE 32817.7200 35705.7100 28509.2500 37316.8100 6-PHOSPHOGLUCONATE (1) 2545.0000 3539.0000 1499.0000 1494.0000 AMINOISOBUTANOATE 360.0000 248.0000 199.0000 193.0000 ANSERINE 47139.0000 42911.0000 39051.0000 38918.0000 ARGININE 65925.0000 69933.0000 45754.0000 43034.0000 ASPARAGINE 4382.0000 3915.0000 1042.0000 1311.0000 AZELATE 8218.8200 9666.9800 10015.3400 9213.3200 BETAINE 22908.0000 24784.0000 13146.0000 16534.0000 CARNOSINE 10408.4600 8354.1300 1212.2000 2021.3400 CDP-ETHANOLAMINE 20060.0000 19825.0000 15278.0000 13965.0000 CIS-4-HYDROXY-D-PROLINE 363.0000 563.0000 520.0000 554.0000 CITICOLINE 3919.4600 2976.7400 4785.3200 5206.6000 CITRAMALATE 152808.8300 205128.0300 150681.8000 162480.0600 CITRATE 1031823.0000 1061641.0000 691807.0000 836070.0000 CORTISONE 8862.2800 8408.8900 2971.9400 2730.9500 CREATINE 1482.0300 1356.0000 854.0000 872.2000 CYSTEATE 162374.0000 260231.0000 154414.0000 72892.0000 CYSTEINE 1638.0000 2430.0000 1290.0000 1354.0000 CYSTEINE (1) 1582.0000 1431.0000 505.0000 797.0000 CYSTINE 15899.0000 28717.0000 14732.0000 14325.0000 D4-TAURINE 250963.0000 329146.1200 305970.7200 283350.0300 DEOXYADENOSINE 40105.6700 49182.4000 44846.8700 44352.9600 DEOXYGUANOSINE 7010.7600 9924.6000 8075.8800 6687.8300 DEOXYGUANOSINE-MONOPHOSPHATE 926608.3800 903649.3100 505776.4400 491006.4400 DETHIOBIOTIN 2722.0000 1515.0000 2869.0000 2130.0000 DIAMINOPIMELATE 16063.0000 9674.0000 5199.0000 3622.0000 DIHYDROXYACETONE PHOSPHATE 6567.0000 21130.0000 5938.0000 2574.0000 D-PINITOL 75594.0000 82929.0000 51274.0000 71875.0000 D-RIBOSE 5-PHOSPHATE 5809.0000 8363.0000 3741.0000 2862.0000 D-SEDOHEPTULOSE 3799.0000 7159.0000 4739.0000 7218.0000 FERULATE 926892.0600 1189551.0000 1380037.0000 871674.1900 FRUCTOSE 6-PHOSPHATE 5928.0000 8323.0000 4208.0000 2973.0000 FRUCTOSE BISPHOSPHATE 45195.0000 55829.0000 26608.0000 24411.0000 FUMARATE 11208.8400 12021.7900 7248.5900 6233.6500 G6P and G1P 29342.0000 34649.0000 15700.0000 16260.0000 GALACTOSE 1-PHOSPHATE 2626.0000 3556.0000 1924.0000 1464.0000 GAMMA-AMINOBUTYRATE 360.0000 248.0000 187.0000 193.0000 GLUCONATE 12749.0000 25240.0000 2052.4300 16339.8200 GLUCOSAMINATE 9608.0400 9149.7800 6701.5000 7654.6800 GLUCOSAMINE 6-PHOSPHATE 223.0000 269.0000 235.0000 182.0000 GLUCOSAMINE 6-SULFATE 617.0000 990.0000 878.0000 478.0000 GLUTAMATE 423134.0000 451532.0000 208361.0000 242007.0000 GLUTAMINE 1268999.0000 906794.0000 1189631.0000 897445.0000 GLUTARATE 29347.2000 68312.5100 37242.8000 42061.6800 GLYCERALDEHYDE 3-PHOSPHATE 9289.0000 16600.0000 3287.0000 2859.0000 GLYCERATE 1350.0000 1457.0000 900.0000 533.0000 GLYCEROL 3-PHOSPHATE 18630.0000 42270.0000 15785.0000 6406.0000 GUANIDINOSUCCINATE 9926.0000 13430.0000 14514.0000 9177.0000 GUANOSINE MONOPHOSPHATE 251969.1600 246653.3300 143794.3800 144248.9700 HEXOES (GLUCOSE) 104018.0000 113846.0000 70181.0000 69720.0000 HISTIDINE 39008.8500 41250.3100 67204.9400 21913.7700 HOMOCYSTEINE 172.0000 291.0000 230.0000 354.0000 HYDROXYPHENYLLACTIC ACID (1) 1287444.1200 1608133.7500 1623987.3800 1319786.7500 INOSINE 67978.0000 71558.1400 49199.7500 54180.0000 INOSINE-MONOPHOSPHATE 100972.7100 106598.9800 57690.5100 65708.0400 ISOCITRATE 60902.0000 67583.0000 36101.0000 38800.0000 ISOLEUCINE 98386.0000 112620.0000 53781.0000 74026.0000 ITACONATE 39655.7100 44681.0000 71919.7300 34488.1000 ITACONATE (1) 140983.9100 178951.8300 215839.0000 147972.9500 LACTATE 162000.6600 159895.0800 126617.4100 131455.5900 L-ALANINE 5096.0000 4492.0000 2489.0000 2747.0000 L-CARNITINE 218.0000 244.0000 90.0000 98.0000 LEUCINE 137604.0000 145658.0000 71069.0000 80096.0000 LYSINE 77981.0000 78115.0000 40420.0000 45412.0000 MALATE 124378.3700 132723.8100 112327.0000 70034.8200 MALEATE 69846.6600 84729.5100 101728.4100 69063.6900 MALTOSE 9221.0000 11209.0000 9579.0000 8395.0000 MANDELATE 344253.7200 426499.1900 428214.0900 346761.9100 MANNITOL 15088.0000 25479.0000 18165.0000 18540.0000 MANNOSE 6-PHOSPHATE 2211.0000 3602.0000 1340.0000 1541.0000 METHIONINE 14510.0000 13970.0000 6254.0000 6982.0000 METHIONINE SULFOXIMINE 1574.0000 1338.0000 728.0000 666.0000 METHYL 4-AMINOBUTYRATE 22908.0000 24784.0000 13146.0000 16534.0000 METHYLGLUTARATE 11799.9600 9690.4300 11050.7000 13696.7800 MEVALONATE 2765.0000 3305.0000 2441.0000 2716.0000 MONOMETHYLGLUTARATE 59712.3400 70209.9500 69457.1700 68591.1600 N-ACETYLALANINE 1691.0000 2235.0000 908.0000 870.0000 N-ACETYLASPARAGINE 21036.0000 22501.0000 22389.0000 19415.0000 N-ACETYLASPARTATE 386503.0000 541326.0000 302366.0000 290443.0000 N-ACETYLGALACTOSAMINE 5030.0000 3958.0000 3837.0000 2362.0000 N-ACETYLGLUTAMATE 33410.0000 31648.0000 18422.0000 20848.0000 N-ACETYLGLYCINE 1439.0000 1490.0000 866.0000 1074.0000 N-ACETYLLEUCINE 4969.5300 3691.1600 2614.8600 2282.3400 N-ACETYLMETHIONINE 60812.2800 49312.6000 6647.3300 11510.0700 N-ACETYLNEURAMINATE 80861.0000 73091.8400 47130.0000 53317.0700 N-ACETYLPHENYLALANINE 6923.4400 5726.5600 4670.3200 5068.8400 N-ACETYLSERINE 8081.0000 9025.0000 5363.0000 4738.0000 N-ACETYLTRYPTOPHAN 293.0000 415.0000 367.0000 264.0000 N-ALPHA-ACETYLLYSINE 9944.0000 8564.0000 3232.0000 2872.0000 NICOTINAMIDE MONONUCLEOTIDE 4846.0600 5326.5300 3724.1800 3459.2000 N-METYL-ALANINE 360.0000 248.0000 199.0000 193.0000 N_N-DIMETHYLARGININE 15798.0000 13021.0000 4176.0000 5696.0000 NORVALINE 22908.0000 24784.0000 13146.0000 16534.0000 O-PHOSPHOETHANOLAMINE 63180.0000 66907.0000 31763.0000 39006.0000 O-PHOSPHOSERINE 2739.0000 2382.0000 1765.0000 1633.0000 OPHTHALMATE 7499.0000 8087.0000 1448.0000 2830.0000 OXOGLUTARATE 15288.3700 27382.1100 20045.9400 22621.5600 PANTOTHENATE 102817.0000 117135.0000 110459.0000 85007.0000 PENTOSE 479.0000 989.0000 254.0000 419.0000 PHENYLALANINE 18919632.0000 23960060.0000 21498572.0000 23440114.0000 PHOSPHONOACETATE 1364.0000 1369.0000 847.0000 911.0000 PIPECOLATE 512.0000 531.0000 658.0000 613.0000 PROLINE 2764.0000 3153.0000 1190.0000 1614.0000 PYRIDOXAL 106887.0000 186169.0000 155308.0000 133424.0000 PYRIDOXINE 77011.7000 99595.1600 80057.8000 74533.9100 PYRUVIC ALDEHYDE 45591.0000 60448.0500 63368.6400 45017.0000 SEBACATE 5478.5100 5582.1300 8073.1100 5723.5300 SERINE 156391.0000 157202.0000 84544.0000 101936.0000 SHIKIMATE 564210.0000 649952.0000 532039.0000 735461.0000 STACHYOSE 4745.0000 5191.0000 3426.0000 3108.0000 SUBERATE 9084.7700 9788.5300 10582.7400 10832.5000 SUCCINATE 10072.0400 22341.4800 18242.8300 9059.3300 SUGAR ALCOHOL (RIBITOL_ XYLITOL_ ARABITOL) 220.6500 2333.7900 TAURINE 67703.0000 103505.0000 41949.0000 42068.0000 THREONINE and HOMOSERINE 25891.0000 39541.0000 17385.0000 17780.0000 TRANS-4-HYDROXY-L-PROLINE 363.0000 563.0000 520.0000 554.0000 TRANS-ACONITATE 13336.0100 12820.2400 14567.6700 6579.1200 TRYPTOPHAN 42626.6800 44010.3200 23876.2700 24266.5300 TYROSINE 148659.0000 163421.0000 84072.0000 104456.0000 URIDINE MONOPHOSPHATE 262123.0000 265535.0000 145026.0000 143195.0000 VALINE 22908.0000 24784.0000 13146.0000 16534.0000 MS_METABOLITE_DATA_END #METABOLITES METABOLITES_START metabolite_name pubchem_id inchi_key kegg_id other_id other_id_type ri ri_type moverz_quant 10-HYDROXYDECANOATE C02774 10.251 187.132828 (2-AMINOETHYL)PHOSPHONATE C03557 1.4 124.016029 2-HYDROXYBUTYRATE C05984 2.436 103.039528 3-(2-HYDROXYPHENYL)PROPANOATE C01198 10.575 165.055267 3,4 DIHYDROXYMANDELATE C05580 6.993 183.029465 3,4-DIHYDROXYPHENYLGLYCOL C05576 6.188 169.051468 3-HYDROXYMETHYLGLUTARATE C03761 4.333 161.04509 3-METHOXY-4-HYDROXYMANDELATE C05584 7.169 197.045319 3-SULFINOALANINE C00606 2.049 152.001724 4-HYDROXYBENZALDEHYDE C00633 10.453 121.028854 5-AMINOLEVULINATE C00430 1.398 130.05043 5-AMINOPENTANOATE C00431 1.635 116.071213 6-CARBOXYHEXANOATE C02656 6.032 159.065643 6-PHOSPHOGLUCONATE (1) C00345 3.659 275.01651 AMINOISOBUTANOATE C05145 1.494 102.055565 ANSERINE C01262 3.25 239.112305 ARGININE C00062 2.861 173.103912 ASPARAGINE C00152 1.506 131.045334 AZELATE C08261 9.674 187.096954 BETAINE C00719 1.635 116.071213 CARNOSINE C00386 3.684 225.098602 CDP-ETHANOLAMINE C00570 5.198 445.0513 CIS-4-HYDROXY-D-PROLINE C03440 1.398 130.05043 CITICOLINE C00307 5.332 487.102753 CITRAMALATE C00815 3.823 147.029648 CITRATE C00158 4.6 191.019287 CORTISONE C00762 13.648 359.190582 CREATINE C00300 3.604 130.061218 CYSTEATE C00506 1.924 167.997147 CYSTEINE C00097 2.039 120.011887 CYSTEINE (1) C00097 1.556 120.011917 CYSTINE C01420 2.039 239.016251 D4-TAURINE D4-TAURINE 1.392 128.033127 DEOXYADENOSINE C00559 8.728 250.097275 DEOXYGUANOSINE C00330 9.096 266.088257 DEOXYGUANOSINE-MONOPHOSPHATE C00362 12.395 346.055237 DETHIOBIOTIN C01909 7.342 213.123856 DIAMINOPIMELATE C00666 1.647 189.087509 DIHYDROXYACETONE PHOSPHATE C00111 2.093 168.990265 D-PINITOL C03844 1.85 193.06955 D-RIBOSE 5-PHOSPHATE C00117 2.4 229.011124 D-SEDOHEPTULOSE C02076 1.817 209.064423 FERULATE C01494 7.599 193.050705 FRUCTOSE 6-PHOSPHATE C00085 2.627 259.021881 FRUCTOSE BISPHOSPHATE FRUCTOSE BISPHOSPHATE 3.717 338.987793 FUMARATE C00122 5.332 115.003044 G6P and G1P G6P and G1P 2.75 259.02182 GALACTOSE 1-PHOSPHATE C00446 2.483 259.021729 GAMMA-AMINOBUTYRATE C00334 1.494 102.055542 GLUCONATE C00257 2.799 195.050644 GLUCOSAMINATE C03752 1.851 194.067657 GLUCOSAMINE 6-PHOSPHATE C00352 1.54 258.038757 GLUCOSAMINE 6-SULFATE C04132 2.097 258.031586 GLUTAMATE C00025 1.853 146.045792 GLUTAMINE C00064 1.699 145.06221 GLUTARATE C00489 4.451 131.034195 GLYCERALDEHYDE 3-PHOSPHATE C00661 1.85 168.991196 GLYCERATE C00258 1.819 105.018715 GLYCEROL 3-PHOSPHATE C00093 1.925 171.005997 GUANIDINOSUCCINATE C03139 3.951 174.049805 GUANOSINE MONOPHOSPHATE C00144 10.099 362.049744 HEXOES (GLUCOSE) HEXOES (GLUCOSE) 1.903 179.055908 HISTIDINE C00135 2.976 154.061203 HOMOCYSTEINE C00155 1.697 134.028946 HYDROXYPHENYLLACTIC ACID (1) C03672 15.239 181.050583 INOSINE C00294 7.961 267.076965 INOSINE-MONOPHOSPHATE C00130 8.388 347.038849 ISOCITRATE C00311 4.443 191.018936 ISOLEUCINE C00407 2.083 130.087021 ITACONATE C00490 4.909 129.018585 ITACONATE (1) C00490 7.977 129.018677 LACTATE C00256 1.959 89.024231 L-ALANINE C00041 1.376 88.03997 L-CARNITINE C00318 1.566 160.097687 LEUCINE C00123 1.947 130.087204 LYSINE C00047 1.284 145.09787 MALATE C00497 3.352 133.013565 MALEATE C01384 6.676 115.003059 MALTOSE C00208 3.921 341.10791 MANDELATE C01984 6.254 151.039429 MANNITOL C00392 1.807 181.071213 MANNOSE 6-PHOSPHATE C00275 2.917 259.02179 METHIONINE C00073 2.326 148.043304 METHIONINE SULFOXIMINE C03510 1.529 179.050644 METHYL 4-AMINOBUTYRATE C00334 1.635 116.07122 METHYLGLUTARATE METHYLGLUTARATE 5.023 145.049927 MEVALONATE C00418 3.669 147.065628 MONOMETHYLGLUTARATE MONOMETHYLGLUTARATE 3.785 145.050079 N-ACETYLALANINE N-ACETYLALANINE 3.913 130.050537 N-ACETYLASPARAGINE N-ACETYLASPARAGINE 3.952 173.055145 N-ACETYLASPARTATE C01042 4.281 174.040604 N-ACETYLGALACTOSAMINE C01074 3.018 220.082382 N-ACETYLGLUTAMATE C00624 4.684 188.056137 N-ACETYLGLYCINE N-ACETYLGLYCINE 3.646 116.034744 N-ACETYLLEUCINE C02710 5.7 172.096954 N-ACETYLMETHIONINE C02712 5.849 190.053543 N-ACETYLNEURAMINATE C19910 3.301 308.097931 N-ACETYLPHENYLALANINE C03519 8.988 206.081833 N-ACETYLSERINE N-ACETYLSERINE 3.648 146.045303 N-ACETYLTRYPTOPHAN N-ACETYLTRYPTOPHAN 1.71 245.090744 N-ALPHA-ACETYLLYSINE C12989 3.148 187.108261 NICOTINAMIDE MONONUCLEOTIDE C00455 4.746 333.048401 N-METYL-ALANINE C02721 1.494 102.055542 N,N-DIMETHYLARGININE C03626 3.727 201.134125 NORVALINE C01799 1.635 116.07122 O-PHOSPHOETHANOLAMINE C00346 1.36 140.011154 O-PHOSPHOSERINE C01005 1.684 184.000931 OPHTHALMATE OPHTHALMATE 4.127 288.1203 OXOGLUTARATE C00026 4.634 145.013596 PANTOTHENATE C00864 5.017 218.102768 PENTOSE PENTOSE 1.733 149.0457 PHENYLALANINE C02057 5.322 164.072189 PHOSPHONOACETATE C05682 3.027 138.97963 PIPECOLATE C00408 1.708 128.070679 PROLINE C00148 1.55 114.055481 PYRIDOXAL C00250 5.389 166.050323 PYRIDOXINE C00314 5.291 168.065903 PYRUVIC ALDEHYDE C00546 6.679 71.013298 SEBACATE C08277 11.105 201.112778 SERINE C00065 1.371 104.035294 SHIKIMATE C00493 4.96 173.04509 STACHYOSE C01613 4.616 665.21405 SUBERATE C08278 7.102 173.081375 SUCCINATE C00042 4.016 117.018669 SUGAR ALCOHOL (RIBITOL, XYLITOL, ARABITOL) SUGAR ALCOHOL (RIBITOL, XYLITOL, ARABITOL) 1.865 151.061554 TAURINE C00245 1.404 124.007118 THREONINE and HOMOSERINE THREONINE and HOMOSERINE 1.424 118.050537 TRANS-4-HYDROXY-L-PROLINE C01157 1.398 130.05043 TRANS-ACONITATE C02341 7.55 173.00856 TRYPTOPHAN C00525 10.034 203.081696 TYROSINE C00082 5.6 180.066086 URIDINE MONOPHOSPHATE C00105 6.715 323.028595 VALINE C00183 1.635 116.071205 METABOLITES_END #END