#METABOLOMICS WORKBENCH juliehaines_20211122_122645 DATATRACK_ID:2940 STUDY_ID:ST001999 ANALYSIS_ID:AN003262 PROJECT_ID:000000 VERSION 1 CREATED_ON November 22, 2021, 12:33 pm #PROJECT PR:PROJECT_TITLE Polyamine import and accumulation causes immunomodulation in macrophages PR:PROJECT_TITLE engulfing apoptotic cells PR:PROJECT_SUMMARY Phagocytosis of apoptotic cells, termed efferocytosis, is critical for tissue PR:PROJECT_SUMMARY homeostasis and drives anti-inflammatory programming in engulfing macrophages. PR:PROJECT_SUMMARY Here, we assess metabolites in naïve and inflammatory macrophages following PR:PROJECT_SUMMARY engulfment of multiple cellular and non-cellular targets. Efferocytosis leads to PR:PROJECT_SUMMARY unique increases in the arginine-derived polyamines, spermidine and spermine, in PR:PROJECT_SUMMARY vitro and in vivo. Surprisingly, polyamine accumulation after efferocytosis does PR:PROJECT_SUMMARY not arise from retention of apoptotic cell metabolites or de novo synthesis, but PR:PROJECT_SUMMARY from enhanced polyamine import that is dependent on Rac1, actin, and PI3 kinase. PR:PROJECT_SUMMARY Blocking polyamine import prevents efferocytosis from suppressing macrophage PR:PROJECT_SUMMARY IL-1beta or IL-6. This identifies efferocytosis as a trigger for polyamine PR:PROJECT_SUMMARY import and accumulation, and imported polyamines as mediators of PR:PROJECT_SUMMARY efferocytosis-induced immune reprogramming. PR:INSTITUTE University of Colorado Denver 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 Polyamine import and accumulation causes immunomodulation in macrophages ST:STUDY_TITLE engulfing apoptotic cells (Part 4) ST:STUDY_SUMMARY Phagocytosis of apoptotic cells, termed efferocytosis, is critical for tissue ST:STUDY_SUMMARY homeostasis and drives anti-inflammatory programming in engulfing macrophages. ST:STUDY_SUMMARY Here, we assess metabolites in naïve and inflammatory macrophages following ST:STUDY_SUMMARY engulfment of multiple cellular and non-cellular targets. Efferocytosis leads to ST:STUDY_SUMMARY unique increases in the arginine-derived polyamines, spermidine and spermine, in ST:STUDY_SUMMARY vitro and in vivo. Surprisingly, polyamine accumulation after efferocytosis does ST:STUDY_SUMMARY not arise from retention of apoptotic cell metabolites or de novo synthesis, but ST:STUDY_SUMMARY from enhanced polyamine import that is dependent on Rac1, actin, and PI3 kinase. ST:STUDY_SUMMARY Blocking polyamine import prevents efferocytosis from suppressing macrophage ST:STUDY_SUMMARY IL-1beta or IL-6. This identifies efferocytosis as a trigger for polyamine ST:STUDY_SUMMARY import and accumulation, and imported polyamines as mediators of ST:STUDY_SUMMARY efferocytosis-induced immune reprogramming. ST:INSTITUTE University of Colorado Denver 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 Mus musculus SU:TAXONOMY_ID 10090 #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 - ctrl 1 Category:unexposed macrophages RAW_FILE_NAME=7+ SUBJECT_SAMPLE_FACTORS - ctrl 2 Category:unexposed macrophages RAW_FILE_NAME=8+ SUBJECT_SAMPLE_FACTORS - ctrl 3 Category:unexposed macrophages RAW_FILE_NAME=9+ SUBJECT_SAMPLE_FACTORS - 1 -AC 8h Category:non-engulfing RAW_FILE_NAME=10+ SUBJECT_SAMPLE_FACTORS - 2 -AC 8h Category:non-engulfing RAW_FILE_NAME=11+ SUBJECT_SAMPLE_FACTORS - 3 -AC 8h Category:non-engulfing RAW_FILE_NAME=12+ SUBJECT_SAMPLE_FACTORS - 1 +AC 8h Category:engulfing RAW_FILE_NAME=13+ SUBJECT_SAMPLE_FACTORS - 2 +AC 8h Category:engulfing RAW_FILE_NAME=14+ SUBJECT_SAMPLE_FACTORS - 3 +AC 8h Category:engulfing RAW_FILE_NAME=15+ #COLLECTION CO:COLLECTION_SUMMARY LPS-primed murine peritoneal macrophages were fed fluorescently-tagged apoptotic CO:COLLECTION_SUMMARY Jurkat cells for 1h. Unengulfed targets were washed off and macrophages were CO:COLLECTION_SUMMARY left to degrade targets for a further 7 hours. Macrophages were then collected CO:COLLECTION_SUMMARY and sorted into non-engulfing or engulfing populations, alongside control CO:COLLECTION_SUMMARY unexposed macrophages. Sorted cells were pelleted at 400xg and dry pellets were CO:COLLECTION_SUMMARY snap frozen and stored at -80C until metabolite extraction. CO:SAMPLE_TYPE Macrophages #TREATMENT TR:TREATMENT_SUMMARY LPS-primed murine peritoneal macrophages were fed fluorescently-tagged apoptotic TR:TREATMENT_SUMMARY Jurkat cells for 1h. Unengulfed targets were washed off and macrophages were TR:TREATMENT_SUMMARY left to degrade targets for a further 7 hours. Macrophages were then collected TR:TREATMENT_SUMMARY and sorted into non-engulfing or engulfing populations, alongside control TR:TREATMENT_SUMMARY unexposed macrophages. Sorted cells were pelleted at 400xg and dry pellets were TR:TREATMENT_SUMMARY snap frozen and stored at -80C until metabolite extraction. #SAMPLEPREP SP:SAMPLEPREP_SUMMARY To process cells for assessment of intracellular metabolites, cells were SP:SAMPLEPREP_SUMMARY pelleted at 400xg in tubes coated with 0.06% BSA. Supernatant was aspirated and SP:SAMPLEPREP_SUMMARY discarded; residual liquid was carefully wicked away from the pellet with a SP:SAMPLEPREP_SUMMARY kimwipe. Dry pellets were immediately snap frozen and stored at -80C until SP:SAMPLEPREP_SUMMARY processing. To process culture supernatants for assessment of metabolites, SP:SAMPLEPREP_SUMMARY supernatant was centrifuged at 400xg to pellet any cells. Cell-free supernatant SP:SAMPLEPREP_SUMMARY was then transferred to a fresh tube, snap frozen, and stored at -80C until SP:SAMPLEPREP_SUMMARY processing. Ultra-high pressure liquid chromatography-mass spectrometry SP:SAMPLEPREP_SUMMARY (UHPLC-MS) was performed by the University of Colorado School of Medicine SP:SAMPLEPREP_SUMMARY Metabolomics Core. Metabolites from frozen cell pellets were extracted at 2e6 SP:SAMPLEPREP_SUMMARY cells/mL in ice cold 5:3:2 MeOH:acetonitrile:water (v/v/v). Media was thawed on SP:SAMPLEPREP_SUMMARY ice and a 10 L aliquot treated with 240 L of the same extraction solution. SP:SAMPLEPREP_SUMMARY Extractions were carried out using vigorous vortexing for 30 min at 4C. SP:SAMPLEPREP_SUMMARY Supernatants were clarified by centrifugation (10 min, 18,000 g, 4C) and SP:SAMPLEPREP_SUMMARY analyzed using a Thermo Vanquish UHPLC coupled to a Thermo Q Exactive mass SP:SAMPLEPREP_SUMMARY spectrometer. #CHROMATOGRAPHY CH:CHROMATOGRAPHY_SUMMARY Isocratic flow at 250 uL/min using 100% phase A (95% water, 5% acetonitrile, 10 CH:CHROMATOGRAPHY_SUMMARY mM ammonium acetate). CH:CHROMATOGRAPHY_TYPE Reversed phase CH:INSTRUMENT_NAME Thermo Vanquish CH:COLUMN_NAME Phenomenex Kinetex C18 (150 x 2.1mm, 2.6 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 Global metabolomics analyses were performed using a 3 min isocratic run in MS:MS_COMMENTS positive and negative ion modes (separate runs) as described previously (Nemkov MS:MS_COMMENTS et al., 2015, Nemkov et al., 2017); stable isotope tracing samples were analyzed MS:MS_COMMENTS using a 5 min C18 gradient in positive and negative ion modes (separate runs) as MS:MS_COMMENTS described (Nemkov et al., 2019, Gehrke et al., 2019). For all analyses, the MS MS:MS_COMMENTS scanned in MS1 mode across the m/z range of 65 to 950. Peaks were annotated (in MS:MS_COMMENTS conjunction with the KEGG database), integrated, and quality control performed MS:MS_COMMENTS using Maven (Princeton University) as described. Stable isotope tracing results MS:MS_COMMENTS were isotopically corrected for the natural abundance of 13C1, 13C2, 15N1, and MS:MS_COMMENTS 15N2 (Nemkov et al., 2017). #MS_METABOLITE_DATA MS_METABOLITE_DATA:UNITS peak area MS_METABOLITE_DATA_START Samples ctrl 1 ctrl 2 ctrl 3 1 -AC 8h 2 -AC 8h 3 -AC 8h 1 +AC 8h 2 +AC 8h 3 +AC 8h Factors Category:unexposed macrophages Category:unexposed macrophages Category:unexposed macrophages Category:non-engulfing Category:non-engulfing Category:non-engulfing Category:engulfing Category:engulfing Category:engulfing ATP 2.87E+04 3.19E+04 2.75E+04 1.61E+03 2.19E+04 6.80E+03 9.91E+03 2.52E+04 6.89E+03 ADP 9.80E+04 6.13E+04 8.52E+04 4.22E+03 2.03E+04 2.61E+03 9.36E+03 7.41E+03 3.88E+04 Adenine 1.80E+05 8.48E+04 1.71E+05 1.10E+05 9.33E+04 8.06E+04 1.24E+05 8.44E+04 1.02E+05 GTP 5.56E+03 7.70E+03 6.64E+03 0.00E+00 8.25E+03 7.21E+03 0.00E+00 7.31E+03 0.00E+00 GDP 4.29E+04 7.67E+04 4.07E+04 1.87E+04 1.76E+04 2.28E+04 7.56E+04 3.93E+04 3.73E+04 Cytidine 0.00E+00 0.00E+00 0.00E+00 0.00E+00 3.19E+03 0.00E+00 1.66E+05 1.15E+05 1.16E+05 UTP 1.67E+04 2.36E+04 2.39E+04 6.39E+03 5.95E+03 1.61E+04 1.48E+04 8.86E+03 5.25E+03 UDP 5.29E+04 6.15E+04 4.66E+04 3.15E+04 2.86E+04 3.01E+04 1.10E+05 5.12E+04 4.54E+04 IDP 3.25E+06 3.03E+06 3.36E+06 4.27E+06 3.37E+06 3.89E+06 3.78E+06 3.64E+06 3.48E+06 Inosine 1.08E+04 8.48E+03 5.82E+03 5.35E+03 1.09E+03 2.00E+04 6.90E+04 4.25E+04 3.68E+04 Xanthine 7.51E+04 6.30E+04 4.03E+04 4.35E+04 3.33E+04 1.96E+04 2.74E+05 2.11E+05 2.12E+05 3',5'-Cyclic IMP 7.96E+06 7.61E+06 8.57E+06 1.45E+07 1.12E+07 1.29E+07 9.49E+06 9.41E+06 9.27E+06 UDP-glucose 1.31E+04 2.17E+04 5.00E+03 7.08E+03 5.70E+03 6.45E+03 3.32E+04 2.11E+04 9.16E+03 Diphosphate 2.07E+07 2.42E+07 2.01E+07 1.22E+07 1.27E+07 1.37E+07 2.23E+07 2.30E+07 1.71E+07 D-Glucose 1.22E+07 1.32E+07 1.51E+07 1.43E+07 1.51E+07 1.71E+07 1.46E+07 1.34E+07 1.69E+07 D-Glucose 6-phosphate 9.23E+04 1.20E+05 5.99E+04 9.64E+04 9.79E+04 5.29E+04 3.25E+05 2.42E+05 1.53E+05 D-Fructose 1-6-bisphosphate 7.07E+05 9.32E+05 5.15E+05 5.55E+05 4.26E+05 3.61E+05 1.73E+06 9.75E+05 6.58E+05 D-Glyceraldehyde 3-phosphate/Glycerone phosphate 3.78E+05 4.75E+05 2.41E+05 3.61E+05 3.05E+05 2.57E+05 9.79E+05 6.60E+05 4.44E+05 2/3-Phospho-D-glycerate 3.12E+05 4.73E+05 2.78E+05 3.76E+05 3.92E+05 3.23E+05 1.01E+06 6.07E+05 4.33E+05 Pyruvate 1.25E+06 1.19E+06 1.51E+06 1.40E+06 1.48E+06 1.49E+06 1.39E+06 1.07E+06 1.45E+06 Lactate 4.72E+06 5.27E+06 6.10E+06 5.52E+06 5.47E+06 6.59E+06 5.88E+06 4.82E+06 6.48E+06 Maltose 1.51E+04 4.68E+03 1.95E+04 2.23E+04 0.00E+00 1.99E+04 2.56E+04 6.23E+03 1.27E+04 Mannitol 1.46E+04 2.94E+04 3.75E+05 9.22E+04 3.69E+04 6.06E+04 1.93E+05 2.54E+04 6.76E+04 D-Ribose 3.31E+05 3.37E+05 4.08E+05 3.91E+05 4.18E+05 4.48E+05 4.06E+05 3.35E+05 4.59E+05 Citrate 1.12E+06 1.54E+06 1.12E+06 1.42E+06 9.16E+05 1.06E+06 1.95E+06 1.56E+06 1.05E+06 2-Oxoglutarate 2.49E+04 3.43E+04 2.66E+04 4.45E+04 3.59E+04 3.51E+04 7.59E+04 3.79E+04 4.22E+04 Succinate 3.17E+05 2.81E+05 3.27E+05 4.08E+05 4.07E+05 4.05E+05 6.89E+05 5.08E+05 3.75E+05 Fumarate 2.72E+04 6.09E+04 5.15E+04 8.88E+04 6.18E+04 5.84E+04 1.30E+05 9.16E+04 8.41E+04 Malate 5.18E+05 6.42E+05 5.19E+05 5.43E+05 3.96E+05 4.27E+05 1.59E+06 1.01E+06 8.23E+05 Itaconate 1.07E+06 1.77E+06 7.65E+05 8.12E+05 6.82E+05 7.66E+05 4.24E+06 3.23E+06 1.97E+06 5-Oxoproline 2.68E+05 2.27E+05 2.35E+05 1.40E+05 1.45E+05 1.50E+05 1.97E+05 1.21E+05 1.92E+05 Ascorbate 1.86E+05 2.03E+05 2.11E+05 4.19E+05 3.04E+05 3.65E+05 2.92E+05 2.57E+05 2.66E+05 UDP-N-acetyl-D-glucosamine 2.83E+04 3.01E+04 9.78E+03 1.31E+04 1.95E+04 1.53E+04 3.49E+04 3.71E+04 2.35E+04 Phosphocreatine 2.72E+04 2.28E+04 2.11E+04 9.18E+03 2.21E+04 2.28E+04 2.59E+04 2.72E+04 2.02E+04 3D-(3-5/4)-Trihydroxycyclohexane-1-2-dione 3.41E+05 3.64E+05 4.42E+05 6.06E+05 5.13E+05 5.48E+05 4.86E+05 4.42E+05 4.90E+05 Glycerol 3-phosphate 3.88E+04 5.49E+04 2.92E+04 5.04E+04 4.42E+04 3.86E+04 9.72E+04 5.40E+04 3.46E+04 Ethanolamine phosphate 5.92E+05 4.48E+05 3.21E+05 1.47E+05 1.88E+05 9.38E+04 3.94E+05 2.61E+05 2.86E+05 Hexanoic acid (caproate) 7.32E+05 6.19E+05 7.41E+05 7.74E+05 7.49E+05 6.39E+05 6.21E+05 6.46E+05 6.74E+05 Heptanoic acid 6.85E+05 4.25E+05 7.00E+05 8.09E+05 7.11E+05 6.99E+05 6.91E+05 5.40E+05 7.24E+05 Octanoic acid (caprylic acid) 1.62E+07 1.12E+07 1.96E+07 6.28E+07 3.16E+07 2.93E+07 3.30E+07 2.10E+07 2.78E+07 Nonanoic acid (pelargonic acid) 9.50E+05 4.18E+05 1.08E+06 2.33E+06 1.01E+06 8.72E+05 1.01E+06 6.53E+05 1.20E+06 Decanoic acid (capric acid) 1.04E+05 4.91E+04 8.84E+04 1.44E+05 9.93E+04 9.37E+04 1.05E+05 7.13E+04 9.48E+04 Dodecanoic acid (lauric acid) 8.20E+04 5.76E+04 9.92E+04 1.31E+05 9.21E+04 8.86E+04 8.96E+04 9.22E+04 7.72E+04 Tetradecanoic acid (myristic acid) 2.23E+05 1.29E+05 2.42E+05 3.31E+05 1.95E+05 3.42E+05 1.97E+05 2.19E+05 1.93E+05 Hexadecanoic acid (palmitic acid) 5.96E+06 2.65E+06 9.34E+06 8.66E+06 6.55E+06 9.34E+06 3.27E+06 6.65E+06 5.85E+06 Octadecanoic acid (stearic acid) 3.95E+06 1.89E+06 6.52E+06 6.18E+06 4.85E+06 6.42E+06 2.69E+06 4.85E+06 4.87E+06 Tetradecenoic acid (myristoleic acid) 1.01E+04 4.19E+03 7.40E+03 2.10E+04 9.89E+03 2.20E+04 9.92E+03 1.48E+04 9.06E+03 Hexadecenoic acid (Palmitoleic acid) 7.64E+04 2.95E+04 5.05E+04 1.07E+05 4.15E+04 1.94E+05 7.44E+04 1.08E+05 5.04E+04 Octadecenoic acid (Oleic acid) 4.23E+05 2.83E+05 6.61E+05 8.76E+05 5.53E+05 8.09E+05 4.45E+05 6.55E+05 4.91E+05 Linoleic acid ((9Z,12Z)-Octadecadienoic acid) 2.14E+05 2.05E+05 2.42E+05 7.33E+05 4.10E+05 3.76E+05 2.94E+05 3.91E+05 2.86E+05 a-Linolenic acid (Octadecatrienoic acid) 1.74E+04 1.89E+04 1.89E+04 7.21E+04 3.89E+04 4.37E+04 4.70E+04 3.34E+04 2.72E+04 Arachidonic acid (Eicosatetraenoic acid) 5.94E+04 5.92E+04 5.46E+04 9.92E+04 6.07E+04 6.27E+04 1.06E+05 8.22E+04 5.35E+04 Adrenic acid (7,10,13,16-Docosatetraenoic acid) 3.14E+04 2.99E+04 2.97E+04 4.87E+04 3.13E+04 3.11E+04 3.82E+04 3.05E+04 2.41E+04 Dihomo-gamma-linolenic acid ((8Z-11Z-14Z)-Icosatrienoic acid) 2.23E+04 2.17E+04 2.74E+04 4.93E+04 2.91E+04 3.63E+04 4.34E+04 4.50E+04 2.63E+04 Docosapentaenoic acid ((7Z-10Z-13Z-16Z-19Z)-Docosa-7-10-13-16-19-pentaenoic acid) 1.44E+04 2.30E+04 2.06E+04 3.36E+04 2.63E+04 2.84E+04 4.18E+04 3.06E+04 2.58E+04 Cholate 2.12E+04 3.05E+04 3.52E+04 1.22E+05 7.16E+04 7.32E+04 3.26E+04 5.74E+04 5.03E+04 Glycochenodeoxycholate 5.10E+04 7.02E+04 6.53E+04 1.88E+05 1.86E+05 1.37E+05 1.04E+05 8.51E+04 7.46E+04 Glycocholate 1.34E+04 2.38E+04 2.86E+04 8.13E+04 7.68E+04 7.19E+04 5.97E+04 4.57E+04 3.33E+04 Taurodeoxycholate 2.27E+04 2.63E+04 2.73E+04 9.03E+04 8.06E+04 5.74E+04 3.95E+04 4.03E+04 3.75E+04 Taurocholate 6.94E+03 8.68E+03 8.75E+03 2.86E+04 2.99E+04 2.76E+04 1.52E+04 1.63E+04 9.40E+03 Prostaglandin D2/Thromboxane A2 (isobars) 1.91E+04 1.04E+04 5.21E+04 1.94E+04 6.43E+04 2.34E+04 1.38E+04 7.28E+04 3.03E+04 Prostaglandin D3 (isobars) 4.62E+03 6.30E+03 5.78E+03 4.83E+03 5.96E+03 1.51E+04 3.92E+03 9.39E+03 6.99E+03 Prostaglandin G2 1.30E+04 6.05E+03 2.31E+04 1.41E+04 2.16E+04 1.20E+04 1.35E+04 1.88E+04 1.24E+04 Cholesterol sulfate 1.58E+04 7.96E+03 1.27E+04 1.37E+04 1.00E+04 1.43E+04 1.77E+04 1.07E+04 1.76E+04 Pentobarbital 3.43E+03 4.81E+03 6.18E+04 5.03E+04 2.09E+04 2.76E+04 1.39E+04 1.26E+04 5.86E+03 2-Methyleneglutarate 1.51E+06 1.66E+06 1.95E+06 1.74E+06 1.82E+06 1.98E+06 1.67E+06 1.46E+06 1.94E+06 6-Pyruvoyltetrahydropterin 3.04E+04 2.24E+04 3.80E+04 6.64E+04 3.94E+04 5.29E+04 5.07E+04 2.85E+04 2.91E+04 D-glucono-1,5-lactone 9.08E+05 9.10E+05 1.16E+06 1.34E+06 1.25E+06 1.31E+06 1.20E+06 9.84E+05 1.19E+06 Catechol 5.26E+05 4.24E+05 4.73E+05 4.84E+05 3.82E+05 4.11E+05 5.53E+05 5.63E+05 4.96E+05 Catechin 7.46E+05 5.29E+05 8.27E+05 1.23E+06 1.10E+06 1.13E+06 2.25E+05 1.17E+06 1.06E+06 D-Ribitol 5-phosphate 6.11E+04 4.67E+04 4.40E+04 1.17E+05 7.35E+04 1.07E+05 9.17E+04 8.89E+04 6.80E+04 Glucosinolate 1.51E+05 3.66E+05 1.43E+05 1.13E+05 1.72E+05 2.14E+05 2.05E+05 2.58E+05 1.19E+05 MS_METABOLITE_DATA_END #METABOLITES METABOLITES_START metabolite_name KEGG ID m/z r.t. ATP C00002 505.9875 0.59 ADP C00008 426.0217 0.58 Adenine C00147 134.0460 0.67 GTP C00044 521.9828 0.58 GDP C00035 442.0164 0.58 Cytidine C00475 242.0782 0.67 UTP C00075 482.9605 0.55 UDP C00015 402.9943 0.56 IDP C00104 427.0068 0.56 Inosine C00294 267.0724 0.65 Xanthine C00385 151.0250 0.68 3',5'-Cyclic IMP C00943 329.0299 0.55 UDP-glucose C00029 565.0470 0.55 Diphosphate C00013 176.9348 0.61 D-Glucose C00031 179.0552 0.64 D-Glucose 6-phosphate C02965 259.0217 0.57 D-Fructose 1-6-bisphosphate C00354 338.9883 0.53 D-Glyceraldehyde 3-phosphate/Glycerone phosphate C00118 168.9895 0.56 2/3-Phospho-D-glycerate C00631 184.9846 0.56 Pyruvate C00022 87.0072 0.65 Lactate C01432 89.0228 0.64 Maltose C00208 341.1086 0.65 Mannitol C00392 181.0708 0.65 D-Ribose C00121 149.0446 0.64 Citrate C00158 191.0186 0.59 2-Oxoglutarate C00026 145.0131 0.58 Succinate C00042 117.0178 0.62 Fumarate C00122 115.0032 0.61 Malate C00149 133.0130 0.58 Itaconate C00490 129.0180 0.60 5-Oxoproline C01879 128.0339 0.62 Ascorbate C00072 175.0240 0.65 UDP-N-acetyl-D-glucosamine C00043 606.0737 0.56 Phosphocreatine C02305 210.0277 0.56 3D-(3-5/4)-Trihydroxycyclohexane-1-2-dione C04287 159.0288 0.65 Glycerol 3-phosphate C00093 171.0052 0.59 Ethanolamine phosphate C00346 140.0105 0.62 Hexanoic acid (caproate) C01585 115.0750 1.73 Heptanoic acid C17714 129.0907 1.86 Octanoic acid (caprylic acid) C06423 143.1064 1.96 Nonanoic acid (pelargonic acid) C01601 157.1221 2.08 Decanoic acid (capric acid) C01571 171.1379 2.16 Dodecanoic acid (lauric acid) C02679 199.1693 2.33 Tetradecanoic acid (myristic acid) C06424 227.2008 2.56 Hexadecanoic acid (palmitic acid) C00249 255.2324 2.93 Octadecanoic acid (stearic acid) C01530 283.2638 3.46 Tetradecenoic acid (myristoleic acid) C08322 225.1852 2.36 Hexadecenoic acid (Palmitoleic acid) C08362 253.2168 2.62 Octadecenoic acid (Oleic acid) C00712 281.2481 2.97 Linoleic acid ((9Z,12Z)-Octadecadienoic acid) C01595 279.2325 2.65 a-Linolenic acid (Octadecatrienoic acid) C06427 277.2168 2.45 Arachidonic acid (Eicosatetraenoic acid) C00219 303.2325 2.55 Adrenic acid (7,10,13,16-Docosatetraenoic acid) C16527 331.2639 2.82 Dihomo-gamma-linolenic acid ((8Z-11Z-14Z)-Icosatrienoic acid) C03242 305.2482 2.73 Docosapentaenoic acid ((7Z-10Z-13Z-16Z-19Z)-Docosa-7-10-13-16-19-pentaenoic acid) C16513 329.2482 2.58 Cholate C00695 407.2815 1.92 Glycochenodeoxycholate C05466 448.3064 1.77 Glycocholate C01921 464.3009 1.74 Taurodeoxycholate C05463 498.2890 1.77 Taurocholate C05122 514.2836 1.74 Prostaglandin D2/Thromboxane A2 (isobars) C00696 351.2201 1.96 Prostaglandin D3 (isobars) C13802 349.2040 1.92 Prostaglandin G2 C05956 367.2140 1.93 Cholesterol sulfate C18043 465.3054 2.01 Pentobarbital C07422 225.1236 1.94 2-Methyleneglutarate C02930 143.0336 0.65 6-Pyruvoyltetrahydropterin C03684 236.0781 1.74 D-glucono-1,5-lactone C00198 177.0396 0.65 Catechol C00090 109.0281 1.86 Catechin C17590 289.0675 0.56 D-Ribitol 5-phosphate C01068 231.0273 0.64 Glucosinolate C17260 316.9843 0.71 METABOLITES_END #END