#METABOLOMICS WORKBENCH QiushiSun_20221128_085601 DATATRACK_ID:3602 STUDY_ID:ST002361 ANALYSIS_ID:AN003855 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 Reverse phase column with positive MS mode CH:INSTRUMENT_NAME SCIEX ExionLC CH:COLUMN_NAME Phenomenex Kinetex F5 Core-shell LC (100 x 2.1 mm,2.6um) CH:COLUMN_TEMPERATURE 30 CH:FLOW_RATE 0.3ml/min CH:SOLVENT_A 95% water/5% acetonitrile; 0.1% formic acid CH:SOLVENT_B 95% acetonitrile/5% water; 0.1% formic acid CH:ANALYTICAL_TIME 20min CH:WASHING_BUFFER 50%Methanol, 50% water CH:CHROMATOGRAPHY_TYPE Reversed phase #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 60-1000. MS:MS_COMMENTS Ion spray voltage = 5000 V; ion source gas 1 (GS1) = 50 psi, ion source gas 2 MS:MS_COMMENTS (GS2) = 50 psi, curtain gas (CUR) = 30 psi, temperature (TEM) = 400 oC. MS:MS_COMMENTS 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 POSITIVE #MS_METABOLITE_DATA MS_METABOLITE_DATA:UNITS peak area MS_METABOLITE_DATA_START Samples Ucp2PomcKO_1_RP_Pos Ucp2PomcKO_2_RP_Pos Ctrl_1_RP_Pos Ctrl_2_RP_Pos Factors Genotype:UCP2 POMC-knockout Genotype:UCP2 POMC-knockout Genotype:wild type Genotype:wild type 1_3-DIAMINOPROPANE 7581.0000 8329.0000 6772.0000 6532.0000 1_4-DIAMINOBUTANE (PUTRESCINE) 1677.0000 1908.0000 953.0000 1080.0000 1-AMINOCYCLOPROPANECARBOXYLATE 27723.0000 18655.0000 24250.0000 14417.2000 1-METHYL-L-HISTIDINE 2521.1000 1920.7500 2246.0500 2622.7100 2_4-DIHYDROXYPTERIDINE 10210.0000 10296.0000 3343.0000 10582.0000 25-HYDROXYCHOLESTEROL 810.9100 1030.0600 794.0000 1405.5200 2-ACETAMIDO-2-DEOXY-BETA-D-GLUCOSYLAMINE 7364.0700 11533.4500 3451.0800 2416.7000 (2-AMINOETHYL)PHOSPHONATE 1787.0000 1486.0000 1126.0000 1091.0000 2-AMINOPHENOL (1) 526.2400 426.2400 502.1900 565.0100 2-OXOBUTANOATE 889.1900 703.0000 237.3000 309.3900 2-PHOSPHOGLYCERATE 5145.5000 5887.4300 4254.3200 4531.5900 3-(2-HYDROXYPHENYL)PROPANOATE 8063.0000 12594.0000 9673.4000 11430.0100 3-(4-HYDROXYPHENYL)PYRUVATE 3052.0000 4267.0000 3912.0000 3693.0000 3-HYDROXYBENZYL ALCOHOL 8365.1700 9781.5600 10064.6700 10721.4400 3-HYDROXYPHENYLACETATE 15618.5700 17067.9900 19824.0800 21123.2100 3-METHOXYTYRAMINE 150140.8100 166023.3900 151063.4500 138611.8100 3-METHOXYTYROSINE 12409.0500 14310.3900 2434.7700 4971.6100 3-METHYLADENINE 586.7800 941.0000 520.0000 517.0000 3-PHOSPHOGLYCERATE 5145.5000 5904.0600 3871.9300 4531.5900 3-SULFINOALANINE 926.0000 464.0000 2165.0000 303.0000 4-ACETAMIDOBUTANOATE 71445.6500 81786.9600 84095.6500 99072.5900 4-GUANIDINOBUTANOATE 1372194.2500 1519240.6200 1178141.5000 1276582.0000 4-HYDROXY-3-METHOXYPHENYLGLYCOL 2761.2000 2459.7700 2587.2300 2968.0300 4-HYDROXYBENZALDEHYDE 6978.0000 4794.6100 5024.0000 4656.0100 4-IMIDAZOLEACETATE 2875.0000 4690.2400 2237.1500 1688.5200 4-METHYLCATECHOL 2863.0900 5995.5400 5366.6900 5589.4400 4-QUINOLINECARBOXYLATE 20603.5100 27339.6400 19538.6700 19927.3100 5-AMINOLEVULINATE 4181.8100 7172.7000 4390.3100 2943.2000 5-AMINOPENTANOATE 176263.7000 179716.2300 132018.3400 160087.4100 5-METHYLCYTOSINE 12712.0000 15698.0000 14428.0000 17573.0000 5-VALEROLACTONE (1) 13754.0800 19717.5400 15807.5700 15594.6000 6-CARBOXYHEXANOATE 3411.4600 3884.0800 3963.4000 3783.4700 7-DEHYDROCHOLESTEROL 2217.0000 2063.0000 912.0000 4117.0000 ACETOACETATE (1) 4146.6000 5751.0000 4514.1500 3526.3700 ACETYLHEXOSAMINE (GLU_ GALA_ MANNOSE) 8362.0000 8559.0000 5429.0000 3279.0000 ADENINE 66567.6000 68846.5200 48836.0000 48172.4000 ADENOSINE 132580.8900 65883.4100 234297.7800 245262.1100 ADENOSINE 2^_3^-CYCLIC PHOSPHATE 130881.0000 87283.0000 96870.3100 123976.9800 ADENOSINE DIPHOSPHATE RIBOSE 318051.0000 305422.0000 200548.0000 181982.0000 ADENOSINE-MONOPHOSPHATE 1576903.5000 1625028.6200 1079844.1200 1094072.1200 AGMATINE SULFATE 8701.9100 8728.6800 11581.0000 9886.0000 ALPHA-HYDROXYISOBUTYRATE 661.0000 400.0000 613.0000 452.0000 AMINOISOBUTANOATE 7312.0000 7295.0000 4660.0000 11301.0000 ANSERINE 26330.2100 27198.0900 19034.4200 17358.2100 ARGININE 252346.0300 238884.0500 187994.0000 173532.0600 ASPARAGINE 11550.0000 8369.0000 4185.2000 5133.0000 ASPARTATE 16673.0000 19813.0000 15244.0000 12211.0000 BENZOATE 94221.9200 69751.4500 86300.0000 87470.0000 BETAINE 176263.7000 179716.2300 132018.3400 160087.4100 BETA-NICOTINAMIDE ADENINE DINUCLEOTIDE 3329.0000 1579.0000 3254.0000 2771.0000 BETA-NICOTINAMIDE ADENINE DINUCLEOTIDE (1) 7213.0000 16237.0000 7247.0000 9504.0000 BIS(2-ETHYLHEXYL)PHTHALATE 69409.2900 72224.4500 95668.0000 105700.0000 CDP-ETHANOLAMINE 1982.0000 1812.0000 932.0000 723.0000 CIS-4-HYDROXY-D-PROLINE 4181.8100 7172.7000 4390.3100 2943.2000 CITICOLINE 4709.0000 4466.0000 4534.0000 3129.0000 CITRATE 5215.0000 8944.0000 3930.0000 4443.0000 CITRULLINE 2535.3600 984.4100 467.0200 931.8100 CREATINE 239864.2000 282576.8400 201536.0000 197123.0000 CREATININE 1176.0000 1083.0000 2330.0000 1223.0000 CYSTEAMINE 838.0000 628.0000 582.0000 1057.0000 CYSTEATE 8096.0000 6306.0000 6698.0000 7140.0000 CYSTEINE 4171.0000 4686.0000 3451.0000 3000.0000 CYSTINE 1848.0100 3382.7100 723.5300 942.2400 CYTIDINE 14750.0000 9705.0000 7058.0000 9591.0000 CYTIDINE 2^_3^-CYCLIC PHOSPHATE 1430.0000 2594.0000 1529.0000 1624.0000 CYTIDINE DIPHOSPHATE 25873.0000 22580.0000 21897.0000 24821.0000 CYTIDINE MONOPHOSPHATE 23708.0000 22653.0000 17256.0000 16538.0000 CYTOSINE 3000.0000 3138.0000 2038.0000 1559.0000 D4-TAURINE 425091.0000 381550.0000 340445.0600 359196.0000 DEHYDROEPIANDROSTERONE SULFATE (DHEAS) 3802.0000 3653.0000 5128.0000 5225.0000 DEOXYADENOSINE 3596.4800 4841.7100 583.2200 3437.1200 DEOXYADENOSINE MONOPHOSPHATE 14847.0000 19822.0000 8989.0000 9547.0000 DEOXYADENOSINE MONOPHOSPHATE (1) 11353.0000 14561.0000 7781.6900 7289.4300 DEOXYCARNITINE 14559.9700 16176.3800 6224.7300 7947.1500 DEOXYCHOLATE 7613.4100 8719.5700 8457.8100 7975.2000 DEOXYCYTIDINE (1) 5169.8200 5380.5600 1641.4800 993.0800 DEOXYGUANOSINE-MONOPHOSPHATE 42028.0700 35176.3900 30394.3900 30912.0900 DEOXYURIDINE-MONOPHOSPHATE 6535.0000 12218.0000 5432.0000 5911.0000 DETHIOBIOTIN 5292.0100 4494.5800 2925.2900 4245.9800 DIAMINOPIMELATE 48539.4100 42821.0700 16817.0000 15336.0600 DIETHYL 2-METHYL-3-OXOSUCCINATE 14647.8500 13744.9100 16445.6000 14238.0000 DIHYDROFOLATE 1908.2800 2410.6700 1396.9400 1358.9200 DIPALMITOYL-PHOSPHOETHANOLAMINE 7587.0000 4994.0300 3254.5000 12889.2000 DOCOSAHEXAENOATE 114202.0000 77607.0000 36461.0000 66141.0000 DOPAMINE 3919.2600 5863.3700 6970.1000 6310.8100 D-SEDOHEPTULOSE 1418.9200 1179.0000 1354.7500 1079.0000 EICOSAPENTAENOATE 6475.0000 5416.0000 2894.0000 3828.0000 EPINEPHRINE 9856.0700 5793.0100 4519.0900 5142.2700 FOLATE 25039.0000 27540.0000 20144.0000 23980.0000 GAMMA-AMINOBUTYRATE 7312.0000 7295.0000 4660.0000 11301.0000 GLUCOSAMINE 6-PHOSPHATE 1437.0000 1155.0000 2557.0000 1169.0000 GLUCOSAMINE 6-SULFATE 1137.0000 1994.0000 2329.0000 1065.0000 GLUCURONATE 400.0000 782.0000 495.0000 350.0000 GLUTAMATE 119684.0300 114321.0000 80430.0000 75742.0000 GLUTAMINE 277325.5000 320957.6200 203479.4500 253940.5900 GLUTATHIONE REDUCED 8116264.0000 7613349.0000 4341968.0000 5316488.0000 GLYCERALDEHYDE 1188.0000 1424.0000 1319.0000 1642.0000 GLYCEROL-MYRISTATE (1) 2087.7000 2769.3000 2573.0000 2383.1900 GLYCINE 2415.0000 1967.0000 3653.0000 2750.0000 GUANIDINOACETATE 13999.6100 12156.9000 7648.5000 8210.0600 GUANINE 151401.8800 144111.2700 104687.9100 108455.1100 GUANOSINE 6439.6000 8441.1700 2930.2700 5514.0900 HEXOESAMINE(GLUCOSAMINE) 1439.0000 1119.0000 1101.0000 1029.0000 HISTIDINE 56715.4300 53351.2300 35220.0000 35486.1100 HOMOCYSTEINE 1212.7500 978.3200 2891.8400 974.5700 HOMOCYSTEINE THIOLACTONE (1) 710.0000 1076.0000 663.0000 910.0000 HYPOTAURINE 6316.0000 4149.0000 3970.0000 4126.0000 HYPOXANTHINE (1) 113508.0000 106862.0000 80555.0000 89064.0000 INDOLE-3-ACETATE 1002.1300 1664.0000 1309.0000 1591.0000 INDOLE-3-ETHANOL 54632.0000 39919.0000 20381.6400 22887.0000 INOSINE 62447.5900 62783.6800 9910.6900 10490.6900 INOSINE-MONOPHOSPHATE 93527.6000 92517.0300 71106.7200 79841.8400 ISOLEUCINE 1331122.1200 1833933.2500 1355860.2500 1321125.2500 KYNURENINE 14521.0800 12040.4600 8780.0800 9977.0800 L-ALANINE 7959.0000 13953.0000 4304.0000 3208.0000 LAURATE 2198.0100 3528.0000 3004.1800 2271.0000 L-CARNITINE 10528.0000 16066.0100 11373.2400 5168.0000 L-DOPA (1) 8055.0000 7171.0000 3224.0000 4269.0000 LEUCINE 2679441.0000 2424834.0000 1808220.0000 1858754.0000 LUMICHROME 20961.0000 24132.0000 14448.0000 17988.0000 LYSINE 63391.7400 76514.2300 48481.3100 47758.0600 MALATE 7472.0000 11350.0000 4930.0000 6227.0000 METHIONINE 322638.4400 315296.3400 164295.6700 177153.3100 METHYLGUANIDINE 3146.2900 2127.1100 2551.8900 1377.3200 METHYLTHIOADENOSINE 76927.0000 92112.0000 59848.0000 69135.0000 MEVALOLACTONE 19938.4500 26747.6300 16869.4600 20391.2600 MEVALONATE (1) 2355.0400 3469.3000 1988.3100 2076.9500 MONOMETHYLGLUTARATE 9483.3700 9867.7600 5465.0000 6498.2400 MYRISTATE 6685.2500 8498.0300 7112.4800 6242.9900 N-ACETYLALANINE 177641.0000 217825.0000 207051.0000 234210.0000 N-ACETYLASPARTATE 36655.0000 38543.0000 33456.0000 38278.0000 N-ACETYLGLUTAMATE 7250.0000 5652.0000 4393.0000 5274.0000 N-ACETYLLEUCINE 1254.2300 2054.8100 887.0000 1167.6800 N-ACETYLMETHIONINE 10380.0000 9321.0000 1903.2300 2224.0000 N-ACETYLPHENYLALANINE (1) 3834.3800 4264.0100 3733.6000 3740.0700 N-ACETYLPUTRESCINE 75311.5900 81267.6600 69209.3700 64724.9500 NICOTINAMIDE 491799.4400 518653.4100 326930.1900 378700.4400 N-METYL-ALANINE 7312.0000 7295.0000 4660.0000 11301.0000 N_N-DIMETHYLARGININE 916.9100 1086.5000 1333.3200 533.1000 N_N_N-TRIMETHYLLYSINE 5175.1100 5945.1800 2800.5600 2767.2300 NORSPERMIDINE 276.0000 1091.0000 1059.0000 378.0000 NORVALINE 176263.7000 179716.2300 132018.3400 160087.4100 O-ACETYLCARNITINE 32100.5500 41098.8000 17058.9500 25150.0800 OLEATE 6220.4500 5536.7700 3059.1100 4883.5700 O-PHOSPHOETHANOLAMINE 5814.0000 10907.0000 3574.0000 4227.0000 OPHTHALMATE 7823.0000 8348.0000 3260.0000 4341.0000 O-SUCCINYL-HOMOSERINE 1174.0000 1088.0000 448.0000 607.0000 OXOGLUTARATE 4515.0000 4523.0000 3185.0000 4091.0000 PALMITATE 49242.0000 53571.0000 53409.0000 44687.0000 PANTOTHENATE 290521.1900 338334.4400 203974.2500 236278.5500 PENTANOATE 7132.5500 10446.8400 8234.0000 7629.0000 PENTOSE 4259.9900 1518.0000 2720.0000 3155.9600 PHENYLALANINE 104000744.0000 110410648.0000 109162792.0000 116391064.0000 PHOSPHOCREATINE 3426.0000 3600.0000 2870.5200 5084.6600 PHOSPHORYLCHOLINE 236165.2300 242485.0200 173692.3400 179659.0600 PIPECOLATE 62654.4500 76962.0000 44956.0000 45738.0000 PROLINE 41742.5100 32635.1100 27342.8000 20034.9100 PYRIDOXAL-PHOSPHATE 4058.5200 5169.0000 4247.8100 4678.0400 PYRIDOXAMINE 9443.0200 14036.2600 7824.6900 9595.8800 PYRIDOXINE 181499.7000 239605.0900 188204.2300 203800.7500 PYROGLUTAMATE 106281.0000 125868.0000 97228.0000 93725.0000 QUINATE 1549.0000 1092.0000 337.0000 617.0000 RAFFINOSE 2909.0000 3379.0000 3490.0000 2954.0000 RETINOATE 4136.3400 5213.4800 4841.7200 5410.4000 RIBOFLAVIN 28592.0000 28942.0000 15570.0000 14103.0000 S-ADENOSYLHOMOCYSTEINE 1787.0000 3407.0000 1942.0000 2012.0000 S-ADENOSYLHOMOCYSTEINE (1) 6173.0000 6127.0000 4131.0000 3618.0000 S-ADENOSYLMETHIONINE 34022.0000 28595.0000 30749.0000 18374.0000 SALSOLINOL 11447.7900 14972.6600 12086.6900 14372.8600 SPERMIDINE 666123.0000 747593.0000 552918.0000 565232.0000 SPERMINE 1079972.0000 1257459.0000 842413.0000 854934.0000 SPHINGANINE 1658.0000 1919.0000 1025.0000 2542.0000 STACHYOSE 2354.0000 1935.0000 3296.0000 1879.0000 STEARATE 41747.0300 37065.1800 32153.9200 28092.8400 SUGAR ALCOHOL (RIBITOL_ XYLITOL_ ARABITOL) 732.1000 433.4200 287.8900 629.7300 TAURINE 6428.0000 9478.0000 6176.0000 4972.0000 THIAMINE MONOPHOSPHATE 1448.0000 803.0000 816.0000 625.0000 THIAMINE PYROPHOSPHATE 1614.0000 1631.0000 1459.0000 1649.0000 THIOUREA 1295.0000 1749.0000 2817.0000 1464.0000 THREONINE and HOMOSERINE 38112.0100 56047.8600 35365.6800 35352.8000 THYROTROPIN RELEASING HORMONE 4432.1700 4467.2000 2911.4500 2729.0000 THYROXINE 6485.0000 10655.0000 3329.0000 4491.0000 TRANS-4-HYDROXY-L-PROLINE 4181.8100 7172.7000 4390.3100 2943.2000 TRIMETHYLAMINE 6429.7300 3888.1100 6056.0400 4422.9500 TRYPTOPHAN 578923.0000 654597.0000 305298.0000 345009.0000 TYROSINE 1257334.6200 1327773.0000 979688.6200 986907.5600 URACIL 2119.0000 3054.0000 1161.0000 1809.0000 UREIDOPROPIONATE 25650.0300 23772.6900 8925.0300 8400.9800 URIDINE MONOPHOSPHATE 112058.7700 89296.0500 65094.8200 61197.2600 VALINE 176263.7000 179716.2300 132018.3400 160087.4100 VANILLIN 32337.0000 24983.3100 28325.9900 27432.5300 XANTHINE 3058.0000 4087.0000 3378.0000 2940.0000 XANTHOSINE 14010.5400 13271.1800 3266.2600 3943.9300 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 1,3-DIAMINOPROPANE C00986 0.649 75.091042 1,4-DIAMINOBUTANE (PUTRESCINE) C02896 0.673 89.106743 1-AMINOCYCLOPROPANECARBOXYLATE C01234 0.747 102.05407 1-METHYL-L-HISTIDINE C01152 0.679 170.093048 2,4-DIHYDROXYPTERIDINE C03212 1.091 165.042511 25-HYDROXYCHOLESTEROL C15519 12.23 403.35495 2-ACETAMIDO-2-DEOXY-BETA-D-GLUCOSYLAMINE 2-ACETAMIDO-2-DEOXY-BETA-D-GLUCOSYLAMINE 0.745 221.11203 (2-AMINOETHYL)PHOSPHONATE C03557 0.739 126.031334 2-AMINOPHENOL (1) C01987 0.991 110.059021 2-OXOBUTANOATE C00109 0.712 103.038597 2-PHOSPHOGLYCERATE 2-PHOSPHOGLYCERATE 0.909 186.998505 3-(2-HYDROXYPHENYL)PROPANOATE C01198 5.792 167.068695 3-(4-HYDROXYPHENYL)PYRUVATE C01179 4.942 181.04805 3-HYDROXYBENZYL ALCOHOL C03351 3.109 125.058083 3-HYDROXYPHENYLACETATE C05593 4.904 153.053024 3-METHOXYTYRAMINE C05587 1.508 168.101318 3-METHOXYTYROSINE 3-METHOXYTYROSINE 1.452 212.090698 3-METHYLADENINE C00913 0.748 150.077484 3-PHOSPHOGLYCERATE C00597 0.909 186.998489 3-SULFINOALANINE C00606 0.724 154.016495 4-ACETAMIDOBUTANOATE C02946 1.139 146.079559 4-GUANIDINOBUTANOATE C01035 0.847 146.091278 4-HYDROXY-3-METHOXYPHENYLGLYCOL C05594 1.803 185.079208 4-HYDROXYBENZALDEHYDE C00633 5.043 123.043152 4-IMIDAZOLEACETATE C02835 0.754 127.049507 4-METHYLCATECHOL C06730 5.269 125.058342 4-QUINOLINECARBOXYLATE C06414 1.239 174.053925 5-AMINOLEVULINATE C00430 0.749 132.065643 5-AMINOPENTANOATE C00431 0.798 118.084778 5-METHYLCYTOSINE C02376 0.78 126.065178 5-VALEROLACTONE (1) C02240 1.308 101.058876 6-CARBOXYHEXANOATE C02656 4.477 161.07959 7-DEHYDROCHOLESTEROL C01164 15.521 385.34436 ACETOACETATE (1) C00164 1.807 103.037895 ACETYLHEXOSAMINE (GLU, GALA, MANNOSE) ACETYLHEXOSAMINE (GLU, GALA, MANNOSE) 0.787 222.096146 ADENINE C00147 0.842 136.060272 ADENOSINE C00212 1.056 268.1026 ADENOSINE 2',3'-CYCLIC PHOSPHATE C02353 0.865 330.06424 ADENOSINE DIPHOSPHATE RIBOSE C00301 1.245 560.076294 ADENOSINE-MONOPHOSPHATE C00020 0.835 348.069336 AGMATINE SULFATE C00179 0.678 131.128922 ALPHA-HYDROXYISOBUTYRATE ALPHA-HYDROXYISOBUTYRATE 0.718 105.054245 AMINOISOBUTANOATE C05145 0.738 104.070053 ANSERINE C01262 0.747 241.127563 ARGININE C00062 0.733 175.117203 ASPARAGINE C00152 0.742 133.05986 ASPARTATE C16433 0.748 134.043625 BENZOATE C00180 5.92 123.042946 BETAINE C00719 0.798 118.084778 BETA-NICOTINAMIDE ADENINE DINUCLEOTIDE C00003 0.731 664.111389 BETA-NICOTINAMIDE ADENINE DINUCLEOTIDE (1) C00003 0.888 664.112427 BIS(2-ETHYLHEXYL)PHTHALATE C03690 14.739 391.282013 CDP-ETHANOLAMINE C00570 0.747 447.067352 CIS-4-HYDROXY-D-PROLINE C03440 0.749 132.065643 CITICOLINE C00307 0.727 489.112762 CITRATE C00158 1.004 193.03273 CITRULLINE C00327 0.739 176.102463 CREATINE C00300 0.752 132.075577 CREATININE C00791 0.729 114.066139 CYSTEAMINE C01678 0.71 78.036987 CYSTEATE C00506 0.799 170.010239 CYSTEINE C00097 0.75 122.026283 CYSTINE C01420 0.681 241.031631 CYTIDINE C02961 0.766 244.091537 CYTIDINE 2',3'-CYCLIC PHOSPHATE C02354 0.727 306.048767 CYTIDINE DIPHOSPHATE C00112 0.861 404.023285 CYTIDINE MONOPHOSPHATE C00055 0.779 324.05722 CYTOSINE C00380 0.787 112.049385 D4-TAURINE D4-TAURINE 0.747 130.047394 DEHYDROEPIANDROSTERONE SULFATE (DHEAS) C04555 10.58 369.176849 DEOXYADENOSINE C00559 1.142 252.106979 DEOXYADENOSINE MONOPHOSPHATE C00360 0.989 332.07309 DEOXYADENOSINE MONOPHOSPHATE (1) C00360 0.852 332.075531 DEOXYCARNITINE C01181 0.781 146.116409 DEOXYCHOLATE C04483 9.643 393.294647 DEOXYCYTIDINE (1) C00881 0.999 228.100296 DEOXYGUANOSINE-MONOPHOSPHATE C00362 0.987 348.068268 DEOXYURIDINE-MONOPHOSPHATE C00365 0.949 309.048676 DETHIOBIOTIN C01909 5.078 215.137375 DIAMINOPIMELATE C00666 0.747 191.101288 DIETHYL 2-METHYL-3-OXOSUCCINATE C04067 7.216 203.089569 DIHYDROFOLATE C00415 4.538 444.162872 DIPALMITOYL-PHOSPHOETHANOLAMINE C00350 15.573 692.518677 DOCOSAHEXAENOATE C06429 12.712 329.244904 DOPAMINE C03758 1.05 154.084793 D-SEDOHEPTULOSE C02076 0.676 211.080307 EICOSAPENTAENOATE C06428 12.295 303.229584 EPINEPHRINE C00788 0.751 184.094833 FOLATE C00504 4.537 442.146881 GAMMA-AMINOBUTYRATE C00334 0.738 104.070053 GLUCOSAMINE 6-PHOSPHATE C00352 0.711 260.05072 GLUCOSAMINE 6-SULFATE C04132 0.707 260.043579 GLUCURONATE C00191 0.707 195.049393 GLUTAMATE C00025 0.747 148.059113 GLUTAMINE C00064 0.747 147.075272 GLUTATHIONE REDUCED C00051 0.965 308.09082 GLYCERALDEHYDE C02154 0.651 91.038567 GLYCEROL-MYRISTATE (1) GLYCEROL-MYRISTATE 11.293 303.250977 GLYCINE C00037 0.743 76.039024 GUANIDINOACETATE C00581 0.743 118.060463 GUANINE C00242 0.97 152.055023 GUANOSINE C00387 1.202 284.096588 HEXOESAMINE(GLUCOSAMINE) HEXOESAMINE(GLUCOSAMINE) 0.709 180.086594 HISTIDINE C00135 0.743 156.075241 HOMOCYSTEINE C00155 0.736 136.04332 HOMOCYSTEINE THIOLACTONE (1) HOMOCYSTEINE THIOLACTONE 0.696 118.03199 HYPOTAURINE C00519 0.752 110.026367 HYPOXANTHINE (1) C00262 0.988 137.04451 INDOLE-3-ACETATE C00954 6.313 176.069672 INDOLE-3-ETHANOL C00955 6.239 162.092941 INOSINE C00294 1.18 269.086243 INOSINE-MONOPHOSPHATE C00130 0.985 349.052765 ISOLEUCINE C00407 1.015 132.100891 KYNURENINE C00328 1.696 209.090439 L-ALANINE C00041 0.746 90.054283 LAURATE C02679 11.151 201.183212 L-CARNITINE C00318 0.752 162.111084 L-DOPA (1) C00355 1.036 198.074677 LEUCINE C00123 1.072 132.100891 LUMICHROME C01727 5.884 243.08548 LYSINE C00047 0.645 147.111298 MALATE C00497 0.966 135.027557 METHIONINE C00073 0.953 150.057053 METHYLGUANIDINE C02294 0.748 74.071068 METHYLTHIOADENOSINE C00170 3.297 298.094482 MEVALOLACTONE MEVALOLACTONE 1.511 131.068909 MEVALONATE (1) C00418 0.747 149.080063 MONOMETHYLGLUTARATE MONOMETHYLGLUTARATE 3.954 147.063339 MYRISTATE C06424 12.177 229.213898 N-ACETYLALANINE N-ACETYLALANINE 1.142 132.064362 N-ACETYLASPARTATE C01042 0.996 176.053864 N-ACETYLGLUTAMATE C00624 1.057 190.069321 N-ACETYLLEUCINE C02710 4.825 174.11145 N-ACETYLMETHIONINE C02712 3.109 192.067245 N-ACETYLPHENYLALANINE (1) C03519 5.316 208.096085 N-ACETYLPUTRESCINE C02714 0.642 131.116165 NICOTINAMIDE C00153 0.994 123.05407 N-METYL-ALANINE C02721 0.738 104.070053 N,N-DIMETHYLARGININE C03626 0.741 203.149368 N,N,N-TRIMETHYLLYSINE C03793 0.701 189.1586 NORSPERMIDINE C03375 0.718 132.149246 NORVALINE C01799 0.798 118.084778 O-ACETYLCARNITINE C02571 0.998 204.121429 OLEATE C00712 13.319 283.2612 O-PHOSPHOETHANOLAMINE C00346 0.769 142.025467 OPHTHALMATE OPHTHALMATE 0.995 290.133514 O-SUCCINYL-HOMOSERINE C01118 0.998 220.080231 OXOGLUTARATE C00026 1.012 147.028229 PALMITATE C00249 13.114 257.245239 PANTOTHENATE C00864 1.752 220.116287 PENTANOATE C00803 4.935 103.074409 PENTOSE PENTOSE 0.725 151.060318 PHENYLALANINE C02057 1.474 166.08638 PHOSPHOCREATINE C02305 0.789 212.041138 PHOSPHORYLCHOLINE C00588 0.747 184.071823 PIPECOLATE C00408 0.645 130.084885 PROLINE C00148 0.753 116.069603 PYRIDOXAL-PHOSPHATE C00018 0.995 248.030563 PYRIDOXAMINE C00534 0.686 169.095627 PYRIDOXINE C00314 0.996 170.079636 PYROGLUTAMATE C01879 0.993 130.048447 QUINATE C06746 0.74 193.070602 RAFFINOSE C00492 0.758 505.175201 RETINOATE C00777 12.51 301.214386 RIBOFLAVIN C00255 4.724 377.14325 S-ADENOSYLHOMOCYSTEINE C00021 0.751 385.129486 S-ADENOSYLHOMOCYSTEINE (1) C00021 0.994 385.127747 S-ADENOSYLMETHIONINE C00019 0.731 399.141815 SALSOLINOL C09642 1.089 180.100266 SPERMIDINE C00315 0.639 146.164276 SPERMINE C00750 0.632 203.222519 SPHINGANINE C00836 10.311 302.303009 STACHYOSE C01613 0.743 667.2276 STEARATE C01530 13.977 285.27655 SUGAR ALCOHOL (RIBITOL, XYLITOL, ARABITOL) SUGAR ALCOHOL (RIBITOL, XYLITOL, ARABITOL) 0.739 153.074814 TAURINE C00245 0.771 126.020988 THIAMINE MONOPHOSPHATE C01081 0.739 345.075134 THIAMINE PYROPHOSPHATE C00068 0.744 425.040985 THIOUREA C14415 0.647 77.016983 THREONINE and HOMOSERINE THREONINE and HOMOSERINE 0.746 120.06456 THYROTROPIN RELEASING HORMONE C03958 0.994 363.179443 THYROXINE C01829 7.817 777.695312 TRANS-4-HYDROXY-L-PROLINE C01157 0.749 132.065643 TRIMETHYLAMINE C00565 0.73 60.08025 TRYPTOPHAN C00525 3.945 205.096085 TYROSINE C00082 1.086 182.079636 URACIL C00106 1.022 113.033653 UREIDOPROPIONATE C02642 0.985 133.059906 URIDINE MONOPHOSPHATE C00105 0.895 325.041901 VALINE C00183 0.798 118.084778 VANILLIN C00755 5.472 153.052948 XANTHINE C00385 1.045 153.039276 XANTHOSINE C01762 1.755 285.085907 METABOLITES_END #END