#METABOLOMICS WORKBENCH ktsumem_20250507_214726 DATATRACK_ID:5895 STUDY_ID:ST003923 ANALYSIS_ID:AN006441 PROJECT_ID:PR002456 VERSION 1 CREATED_ON May 19, 2025, 7:42 pm #PROJECT PR:PROJECT_TITLE Sex difference in BAT thermogenesis depends on PGC-1α–mediated phospholipid PR:PROJECT_TITLE synthesis in mice PR:PROJECT_SUMMARY Brown adipose tissue (BAT), a thermogenic tissue that plays an important role in PR:PROJECT_SUMMARY systemic energy expenditure, has histological and functional sex differences. PR:PROJECT_SUMMARY BAT thermogenic activity is higher in female mice than in male mice. However, PR:PROJECT_SUMMARY the molecular mechanism underlying this functional sex difference has not been PR:PROJECT_SUMMARY fully elucidated. Herein, we demonstrate the role and mechanism of PGC-1α in PR:PROJECT_SUMMARY this sex difference. Inducible adipocyte-specific PGC-1α knockout (KO) mice PR:PROJECT_SUMMARY display mitochondrial morphological defects and decreased BAT thermogenesis only PR:PROJECT_SUMMARY in females. Expression of carbohydrate response-element binding protein beta PR:PROJECT_SUMMARY (Chrebpβ) and its downstream de novo lipogenesis (DNL)-related genes are both PR:PROJECT_SUMMARY reduced only in female KO mice. BAT-specific knockdown of ChREBPβ displays PR:PROJECT_SUMMARY decreased DNL-related gene expression and mitochondrial morphological defects PR:PROJECT_SUMMARY followed by reduced BAT thermogenesis in female wild-type mice. Lipidomics PR:PROJECT_SUMMARY reveals that, PGC-1α increases ether-linked phosphatidylethanolamine (PE) and PR:PROJECT_SUMMARY cardiolipin(18:2)₄ levels through Chrebpβ-dependent and -independent PR:PROJECT_SUMMARY mechanisms in female BAT. Furthermore, PGC-1α enhances the sensitivity of PR:PROJECT_SUMMARY female BAT estrogen signaling, thereby increasing Chrebpβ and its downstream PR:PROJECT_SUMMARY DNL-related gene expression. These findings demonstrate that PGC-1α–mediated PR:PROJECT_SUMMARY phospholipid synthesis plays a pivotal role in BAT thermogenesis in a PR:PROJECT_SUMMARY sex-dependent manner. PR:INSTITUTE Institute of Science Tokyo PR:LAST_NAME Tsujimoto PR:FIRST_NAME Kazutaka PR:ADDRESS 1-5-45 Yushima, Bunkyo-ku, Tokyo, Tokyo, 113-8510, Japan PR:EMAIL ktsumem@tmd.ac.jp PR:PHONE +81-3-5803-5216 #STUDY ST:STUDY_TITLE Sex difference in BAT thermogenesis depends on PGC-1α–mediated phospholipid ST:STUDY_TITLE synthesis in mice ST:STUDY_SUMMARY Brown adipose tissue (BAT), a thermogenic tissue that plays an important role in ST:STUDY_SUMMARY systemic energy expenditure, has histological and functional sex differences. ST:STUDY_SUMMARY BAT thermogenic activity is higher in female mice than in male mice. However, ST:STUDY_SUMMARY the molecular mechanism underlying this functional sex difference has not been ST:STUDY_SUMMARY fully elucidated. Herein, we demonstrate the role and mechanism of PGC-1α in ST:STUDY_SUMMARY this sex difference. Inducible adipocyte-specific PGC-1α knockout (KO) mice ST:STUDY_SUMMARY display mitochondrial morphological defects and decreased BAT thermogenesis only ST:STUDY_SUMMARY in females. Expression of carbohydrate response-element binding protein beta ST:STUDY_SUMMARY (Chrebpβ) and its downstream de novo lipogenesis (DNL)-related genes are both ST:STUDY_SUMMARY reduced only in female KO mice. BAT-specific knockdown of ChREBPβ displays ST:STUDY_SUMMARY decreased DNL-related gene expression and mitochondrial morphological defects ST:STUDY_SUMMARY followed by reduced BAT thermogenesis in female wild-type mice. Lipidomics ST:STUDY_SUMMARY reveals that, PGC-1α increases ether-linked phosphatidylethanolamine (PE) and ST:STUDY_SUMMARY cardiolipin(18:2)₄ levels through Chrebpβ-dependent and -independent ST:STUDY_SUMMARY mechanisms in female BAT. Furthermore, PGC-1α enhances the sensitivity of ST:STUDY_SUMMARY female BAT estrogen signaling, thereby increasing Chrebpβ and its downstream ST:STUDY_SUMMARY DNL-related gene expression. These findings demonstrate that PGC-1α–mediated ST:STUDY_SUMMARY phospholipid synthesis plays a pivotal role in BAT thermogenesis in a ST:STUDY_SUMMARY sex-dependent manner. ST:INSTITUTE Institute of Science Tokyo ST:LAST_NAME Tsujimoto ST:FIRST_NAME Kazutaka ST:ADDRESS 1-5-45 Yushima, Bunkyo-ku, Tokyo, Tokyo, 113-8510, Japan ST:EMAIL ktsumem@tmd.ac.jp ST:PHONE +81-3-5803-5216 #SUBJECT SU:SUBJECT_TYPE Mammal SU:SUBJECT_SPECIES Mus musculus SU:TAXONOMY_ID 10090 SU:GENDER Male and female #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 - M1 Sample source:BAT | Sex:Male | Genotype:Control RAW_FILE_NAME(Raw Data File Name C)=TMDUM030-1_C_20220315_001_d1.mzML; RAW_FILE_NAME(Raw Data File Name O)=TMDUM030-1_O_20220310_001_d3.mzML; RAW_FILE_NAME(Raw Data File Name S)=TMDUM030-1_S_20220310_001_d3.mzML SUBJECT_SAMPLE_FACTORS - M2 Sample source:BAT | Sex:Male | Genotype:Control RAW_FILE_NAME(Raw Data File Name C)=TMDUM030-1_C_20220315_002_d1.mzML; RAW_FILE_NAME(Raw Data File Name O)=TMDUM030-1_O_20220310_002_d3.mzML; RAW_FILE_NAME(Raw Data File Name S)=TMDUM030-1_S_20220310_002_d3.mzML SUBJECT_SAMPLE_FACTORS - M3 Sample source:BAT | Sex:Male | Genotype:Control RAW_FILE_NAME(Raw Data File Name C)=TMDUM030-1_C_20220315_003_d1.mzML; RAW_FILE_NAME(Raw Data File Name O)=TMDUM030-1_O_20220310_003_d3.mzML; RAW_FILE_NAME(Raw Data File Name S)=TMDUM030-1_S_20220310_003_d3.mzML SUBJECT_SAMPLE_FACTORS - M4 Sample source:BAT | Sex:Male | Genotype:Control RAW_FILE_NAME(Raw Data File Name C)=TMDUM030-1_C_20220315_004_d1.mzML; RAW_FILE_NAME(Raw Data File Name O)=TMDUM030-1_O_20220310_004_d3.mzML; RAW_FILE_NAME(Raw Data File Name S)=TMDUM030-1_S_20220310_004_d3.mzML SUBJECT_SAMPLE_FACTORS - M5 Sample source:BAT | Sex:Male | Genotype:Control RAW_FILE_NAME(Raw Data File Name C)=TMDUM030-1_C_20220315_005_d1.mzML; RAW_FILE_NAME(Raw Data File Name O)=TMDUM030-1_O_20220310_005_d3.mzML; RAW_FILE_NAME(Raw Data File Name S)=TMDUM030-1_S_20220310_005_d3.mzML SUBJECT_SAMPLE_FACTORS - MC1 Sample source:BAT | Sex:Male | Genotype:KO RAW_FILE_NAME(Raw Data File Name C)=TMDUM030-1_C_20220315_006_d1.mzML; RAW_FILE_NAME(Raw Data File Name O)=TMDUM030-1_O_20220310_006_d3.mzML; RAW_FILE_NAME(Raw Data File Name S)=TMDUM030-1_S_20220310_006_d3.mzML SUBJECT_SAMPLE_FACTORS - MC2 Sample source:BAT | Sex:Male | Genotype:KO RAW_FILE_NAME(Raw Data File Name C)=TMDUM030-1_C_20220315_007_d1.mzML; RAW_FILE_NAME(Raw Data File Name O)=TMDUM030-1_O_20220310_007_d3.mzML; RAW_FILE_NAME(Raw Data File Name S)=TMDUM030-1_S_20220310_007_d3.mzML SUBJECT_SAMPLE_FACTORS - MC3 Sample source:BAT | Sex:Male | Genotype:KO RAW_FILE_NAME(Raw Data File Name C)=TMDUM030-1_C_20220315_008_d1.mzML; RAW_FILE_NAME(Raw Data File Name O)=TMDUM030-1_O_20220310_008_d3.mzML; RAW_FILE_NAME(Raw Data File Name S)=TMDUM030-1_S_20220310_008_d3.mzML SUBJECT_SAMPLE_FACTORS - MC4 Sample source:BAT | Sex:Male | Genotype:KO RAW_FILE_NAME(Raw Data File Name C)=TMDUM030-1_C_20220315_009_d1.mzML; RAW_FILE_NAME(Raw Data File Name O)=TMDUM030-1_O_20220310_009_d3.mzML; RAW_FILE_NAME(Raw Data File Name S)=TMDUM030-1_S_20220310_009_d3.mzML SUBJECT_SAMPLE_FACTORS - MC5 Sample source:BAT | Sex:Male | Genotype:KO RAW_FILE_NAME(Raw Data File Name C)=TMDUM030-1_C_20220315_010_d1.mzML; RAW_FILE_NAME(Raw Data File Name O)=TMDUM030-1_O_20220310_010_d3.mzML; RAW_FILE_NAME(Raw Data File Name S)=TMDUM030-1_S_20220310_010_d3.mzML SUBJECT_SAMPLE_FACTORS - F1 Sample source:BAT | Sex:Female | Genotype:Control RAW_FILE_NAME(Raw Data File Name C)=TMDUM030-1_C_20220315_011_d1.mzML; RAW_FILE_NAME(Raw Data File Name O)=TMDUM030-1_O_20220310_011_d3.mzML; RAW_FILE_NAME(Raw Data File Name S)=TMDUM030-1_S_20220310_011_d3.mzML SUBJECT_SAMPLE_FACTORS - F2 Sample source:BAT | Sex:Female | Genotype:Control RAW_FILE_NAME(Raw Data File Name C)=TMDUM030-1_C_20220315_012_d1.mzML; RAW_FILE_NAME(Raw Data File Name O)=TMDUM030-1_O_20220310_012_d3.mzML; RAW_FILE_NAME(Raw Data File Name S)=TMDUM030-1_S_20220310_012_d3.mzML SUBJECT_SAMPLE_FACTORS - F3 Sample source:BAT | Sex:Female | Genotype:Control RAW_FILE_NAME(Raw Data File Name C)=TMDUM030-1_C_20220315_013_d1.mzML; RAW_FILE_NAME(Raw Data File Name O)=TMDUM030-1_O_20220310_013_d3.mzML; RAW_FILE_NAME(Raw Data File Name S)=TMDUM030-1_S_20220310_013_d3.mzML SUBJECT_SAMPLE_FACTORS - F4 Sample source:BAT | Sex:Female | Genotype:Control RAW_FILE_NAME(Raw Data File Name C)=TMDUM030-1_C_20220315_014_d1.mzML; RAW_FILE_NAME(Raw Data File Name O)=TMDUM030-1_O_20220310_014_d3.mzML; RAW_FILE_NAME(Raw Data File Name S)=TMDUM030-1_S_20220310_014_d3.mzML SUBJECT_SAMPLE_FACTORS - F5 Sample source:BAT | Sex:Female | Genotype:Control RAW_FILE_NAME(Raw Data File Name C)=TMDUM030-1_C_20220315_015_d1.mzML; RAW_FILE_NAME(Raw Data File Name O)=TMDUM030-1_O_20220310_015_d3.mzML; RAW_FILE_NAME(Raw Data File Name S)=TMDUM030-1_S_20220310_015_d3.mzML SUBJECT_SAMPLE_FACTORS - FC1 Sample source:BAT | Sex:Female | Genotype:KO RAW_FILE_NAME(Raw Data File Name C)=TMDUM030-1_C_20220315_016_d1.mzML; RAW_FILE_NAME(Raw Data File Name O)=TMDUM030-1_O_20220310_016_d3.mzML; RAW_FILE_NAME(Raw Data File Name S)=TMDUM030-1_S_20220310_016_d3.mzML SUBJECT_SAMPLE_FACTORS - FC2 Sample source:BAT | Sex:Female | Genotype:KO RAW_FILE_NAME(Raw Data File Name C)=TMDUM030-1_C_20220315_017_d1.mzML; RAW_FILE_NAME(Raw Data File Name O)=TMDUM030-1_O_20220310_017_d3.mzML; RAW_FILE_NAME(Raw Data File Name S)=TMDUM030-1_S_20220310_017_d3.mzML SUBJECT_SAMPLE_FACTORS - FC3 Sample source:BAT | Sex:Female | Genotype:KO RAW_FILE_NAME(Raw Data File Name C)=TMDUM030-1_C_20220315_018_d1.mzML; RAW_FILE_NAME(Raw Data File Name O)=TMDUM030-1_O_20220310_018_d3.mzML; RAW_FILE_NAME(Raw Data File Name S)=TMDUM030-1_S_20220310_018_d3.mzML SUBJECT_SAMPLE_FACTORS - FC4 Sample source:BAT | Sex:Female | Genotype:KO RAW_FILE_NAME(Raw Data File Name C)=TMDUM030-1_C_20220315_019_d1.mzML; RAW_FILE_NAME(Raw Data File Name O)=TMDUM030-1_O_20220310_019_d3.mzML; RAW_FILE_NAME(Raw Data File Name S)=TMDUM030-1_S_20220310_019_d3.mzML SUBJECT_SAMPLE_FACTORS - FC5 Sample source:BAT | Sex:Female | Genotype:KO RAW_FILE_NAME(Raw Data File Name C)=TMDUM030-1_C_20220315_020_d1.mzML; RAW_FILE_NAME(Raw Data File Name O)=TMDUM030-1_O_20220310_020_d3.mzML; RAW_FILE_NAME(Raw Data File Name S)=TMDUM030-1_S_20220310_020_d3.mzML #COLLECTION CO:COLLECTION_SUMMARY All mice were treated with norepinephrine (NE) and exposed to a warm environment CO:COLLECTION_SUMMARY (33°C) for 30 minutes prior to tissue collection, to evaluate brown adipose CO:COLLECTION_SUMMARY tissue (BAT) metabolism under conditions of maximal oxygen consumption. Tissues CO:COLLECTION_SUMMARY were collected from mice following euthanasia. Brown adipose tissue was isolated CO:COLLECTION_SUMMARY and immediately frozen in liquid nitrogen before being stored at -80°C for CO:COLLECTION_SUMMARY later analysis. CO:SAMPLE_TYPE Brown adipose tissue #TREATMENT TR:TREATMENT_SUMMARY Tamoxifen-inducible adipocyte-specific PGC-1α knockout (KO) mice were generated TR:TREATMENT_SUMMARY by crossing Pgc1a flox/flox mice (JAX stock #009666) with Adipoq-CreERT2 mice TR:TREATMENT_SUMMARY (JAX stock #025124). Eight-week-old male and female KO and floxed Control mice TR:TREATMENT_SUMMARY were injected intraperitoneally with tamoxifen (100 mg/kg) for 5 consecutive TR:TREATMENT_SUMMARY days and analyzed at least 8 weeks after the last injection. All mice were TR:TREATMENT_SUMMARY maintained at 30°C for one week before NE injection and tissue sampling. At the TR:TREATMENT_SUMMARY time of sacrifice, mice were placed at 33°C and injected with norepinephrine TR:TREATMENT_SUMMARY (NE, 1 mg/kg, i.p.). Thirty minutes after NE administration, brown adipose TR:TREATMENT_SUMMARY tissue (BAT) was collected for metabolomic analysis. #SAMPLEPREP SP:SAMPLEPREP_SUMMARY Approximately 25–30 mg of frozen BAT was homogenized with zirconia beads in SP:SAMPLEPREP_SUMMARY 50% acetonitrile containing internal standards. After centrifugation, the SP:SAMPLEPREP_SUMMARY aqueous layer was filtered through a 5-kDa cutoff filter, dried under vacuum, SP:SAMPLEPREP_SUMMARY and reconstituted in Milli-Q water for metabolomic analysis. #CHROMATOGRAPHY CH:CHROMATOGRAPHY_SUMMARY Fused silica capillary (50 μm i.d. × 80 cm). H3302-1021 is designed for the CH:CHROMATOGRAPHY_SUMMARY separation of anionic metabolites such as organic acids and phosphorylated CH:CHROMATOGRAPHY_SUMMARY compounds, under basic conditions. CH:CHROMATOGRAPHY_TYPE CE CH:INSTRUMENT_NAME Agilent 7100 CE CH:COLUMN_NAME Agilent Fused silica capillary (80cm x 50um) CH:SOLVENT_A HMT electrophoresis buffer H3302-1021 CH:SOLVENT_B N/A CH:FLOW_GRADIENT N/A CH:FLOW_RATE N/A CH:COLUMN_TEMPERATURE N/A #ANALYSIS AN:ANALYSIS_TYPE MS #MS MS:INSTRUMENT_NAME Agilent 6230 TOF MS:INSTRUMENT_TYPE TOF MS:MS_TYPE ESI MS:ION_MODE NEGATIVE MS:MS_COMMENTS CE-MS/MS in negative ion mode using dynamic MRM. Peaks extracted using MS:MS_COMMENTS MasterHands and MassHunter (Agilent) and annotated based on m/z and migration MS:MS_COMMENTS time using HMT metabolite database. #MS_METABOLITE_DATA MS_METABOLITE_DATA:UNITS arbitrary unit MS_METABOLITE_DATA_START Samples M1 M2 M3 M4 M5 MC1 MC2 MC3 MC4 MC5 F1 F2 F3 F4 F5 FC1 FC2 FC3 FC4 FC5 Factors Sample source:BAT | Sex:Male | Genotype:Control Sample source:BAT | Sex:Male | Genotype:Control Sample source:BAT | Sex:Male | Genotype:Control Sample source:BAT | Sex:Male | Genotype:Control Sample source:BAT | Sex:Male | Genotype:Control Sample source:BAT | Sex:Male | Genotype:KO Sample source:BAT | Sex:Male | Genotype:KO Sample source:BAT | Sex:Male | Genotype:KO Sample source:BAT | Sex:Male | Genotype:KO Sample source:BAT | Sex:Male | Genotype:KO Sample source:BAT | Sex:Female | Genotype:Control Sample source:BAT | Sex:Female | Genotype:Control Sample source:BAT | Sex:Female | Genotype:Control Sample source:BAT | Sex:Female | Genotype:Control Sample source:BAT | Sex:Female | Genotype:Control Sample source:BAT | Sex:Female | Genotype:KO Sample source:BAT | Sex:Female | Genotype:KO Sample source:BAT | Sex:Female | Genotype:KO Sample source:BAT | Sex:Female | Genotype:KO Sample source:BAT | Sex:Female | Genotype:KO NAD+ 664 676 679 777 770 541 813 523 619 641 884 1109 996 977 901 765 802 769 693 977 cAMP 1.4 1.3 1.3 2 1.6 1.2 1.5 1.1 1.8 1.6 3 2.3 1.7 2.3 1.9 2.6 2 3 1.8 2.7 cGMP N.D. N.D. N.D. N.D. N.D. N.D. N.D. N.D. N.D. N.D. N.D. N.D. N.D. 0.03 N.D. N.D. N.D. N.D. N.D. N.D. NADH 14 15 16 12 15 12 13 12 12 12 16 25 19 16 16 10 13 12 13 13 Xanthine 10 9.6 9.2 7.9 12 12 14 8.4 11 16 18 15 14 14 12 12 19 11 17 19 ADP-ribose 165 194 149 209 173 176 221 159 193 188 252 235 259 256 236 238 240 260 217 283 Mevalonic acid N.D. N.D. N.D. N.D. N.D. N.D. N.D. N.D. N.D. N.D. N.D. N.D. N.D. N.D. N.D. N.D. N.D. N.D. N.D. N.D. UDP-glucose 37 39 38 31 46 34 33 49 40 33 36 46 37 40 30 33 24 39 37 63 Uric acid 25 37 26 32 27 23 40 23 32 43 47 36 39 35 42 36 56 33 33 43 NADP+ 47 55 57 60 54 42 56 37 41 39 54 66 67 52 58 43 43 36 47 54 IMP 150 127 138 102 127 156 151 75 133 110 419 143 242 524 293 211 235 297 383 388 Sedoheptulose 7-phosphate N.D. N.D. N.D. 6 N.D. N.D. N.D. N.D. N.D. N.D. N.D. N.D. N.D. N.D. N.D. 6.8 3.5 1.6 N.D. 1.4 Glucose 6-phosphate 12 9.7 42 88 39 23 50 15 81 106 46 70 28 122 48 91 129 91 36 94 Fructose 6-phosphate 4.8 4.7 15 28 16 8.9 18 5.1 27 36 18 27 9.3 40 18 31 41 34 15 34 Fructose 1-phosphate N.D. N.D. N.D. 1.3 N.D. N.D. N.D. N.D. 8.7 2.8 4.5 N.D. N.D. N.D. N.D. 11 7.1 19 N.D. 12 Galactose 1-phosphate 0.3 0.7 0.5 1.2 0.6 0.01 0.2 0.5 1 1 1.7 0.4 1 1.9 0.9 0.9 1.4 1.3 0.4 2 Glucose 1-phosphate 2.1 1.6 4.3 6.2 3.5 4.1 4.7 1.9 7.8 8.3 4.7 6.9 2.1 8.3 3.6 5.6 8.5 7.3 3.8 7 Acetoacetyl CoA N.D. N.D. N.D. N.D. N.D. N.D. N.D. N.D. N.D. N.D. N.D. 0.12 N.D. N.D. N.D. N.D. N.D. N.D. N.D. 0.07 Acetyl CoA 31 31 37 23 35 20 33 26 39 25 64 68 54 62 30 15 32 22 39 37 Folic acid N.D. N.D. N.D. N.D. N.D. N.D. N.D. N.D. N.D. N.D. N.D. N.D. N.D. N.D. N.D. N.D. N.D. N.D. N.D. N.D. Ribose 5-phosphate 8.2 7.8 6.3 8.4 9.1 7.1 11 6.2 9.8 9.1 13 12 13 12 12 13 13 15 11 13 CoA 161 207 215 322 247 199 204 148 177 163 240 254 172 295 247 259 224 248 209 310 Ribose 1-phosphate 30 30 25 27 31 32 42 20 33 39 43 32 37 39 37 39 48 38 39 42 Ribulose 5-phosphate 3.7 4.2 4 6 7 4.3 7.7 4 6.4 5.8 9.3 7.8 8.2 9.5 8.6 8.7 7.5 9.1 6.9 9 Xylulose 5-phosphate N.D. N.D. N.D. 0.2 N.D. N.D. N.D. N.D. N.D. N.D. 0.9 N.D. N.D. N.D. N.D. 2.9 1.7 4.6 N.D. 2.3 Erythrose 4-phosphate 0.14 N.D. 0.1 0.6 N.D. 0.1 0.4 0.14 0.4 0.6 0.2 0.2 0.3 0.6 0.5 0.7 0.3 1.2 0.3 0.8 HMG CoA 0.3 0.3 0.3 0.3 0.3 0.2 0.3 0.3 0.3 0.4 0.3 0.4 0.3 0.3 0.2 0.3 0.2 0.2 0.2 0.3 Glyceraldehyde 3-phosphate 3.2 3.3 4.4 15 3.7 3.6 9.7 5.9 17 11 21 8.3 10 11 13 14 14 28 13 19 NADPH 35 35 35 31 35 28 28 30 26 27 25 39 31 26 32 17 24 19 25 22 Malonyl CoA 0.3 0.3 0.2 0.2 0.2 0.2 0.2 0.2 0.2 0.2 0.2 0.5 0.3 0.2 0.2 0.2 0.3 0.2 0.2 0.3 Phosphocreatine 9.1 8.7 9.9 23 9.3 11 3.9 11 12 0.8 6.9 1.7 8.6 2.3 3.2 42 5.5 29 15 30 XMP 0.6 0.2 0.13 0.04 0.7 0.4 0.5 0.03 0.05 0.02 0.8 0.4 0.8 0.9 0.6 0.3 0.2 0.3 1.1 0.5 Dihydroxyacetone phosphate 108 89 97 300 129 87 212 110 261 237 369 223 215 266 243 287 273 437 235 310 Adenylosuccinic acid 51 56 39 11 60 56 44 41 41 44 20 76 70 19 31 8.8 22 9.7 26 15 Fructose 1,6-diphosphate 8.3 5.6 10 67 14 8.5 41 12 64 44 101 31 36 51 43 87 53 163 43 121 6-Phosphogluconic acid 3 3.1 2.8 5.4 3.1 3.5 3.6 2.1 7.7 3.7 5.8 4.3 6.2 4.2 4.8 7.2 7.6 5.6 3 6.6 N-Carbamoylaspartic acid 0.07 0.06 0.07 0.11 0.2 0.04 0.14 0.04 0.2 0.12 0.13 0.3 0.4 0.09 0.2 0.2 0.2 0.2 0.11 0.15 PRPP 0.7 0.5 0.8 3.1 1.4 0.6 1.2 0.6 2.4 1.4 3.4 4.8 2.7 5.9 1.8 4.2 2.6 11 5.2 7.3 2-Phosphoglyceric acid 0.4 0.5 0.5 0.6 0.6 0.5 0.7 0.5 0.7 1 0.6 0.9 0.8 1.1 0.9 0.5 0.9 0.9 0.6 0.7 2,3-Diphosphoglyceric acid 51 75 60 54 58 40 73 46 60 53 63 89 91 71 78 64 78 65 52 65 3-Phosphoglyceric acid 2.7 3.2 3 3.4 3.9 2.5 5.5 2.5 4.8 6.4 4.3 7 5.9 8.9 5.8 3.5 6.2 5.4 3.6 5.2 Phosphoenolpyruvic acid N.D. N.D. N.D. N.D. N.D. N.D. N.D. N.D. N.D. N.D. N.D. N.D. N.D. 0.05 N.D. N.D. N.D. N.D. N.D. N.D. GMP 140 201 188 153 151 150 140 170 151 176 149 125 84 115 114 115 112 116 118 155 AMP 1008 1158 1152 932 1278 875 999 945 889 867 1008 1293 1181 987 966 837 856 880 876 1076 2-Oxoisovaleric acid N.D. 4.8 N.D. N.D. N.D. 4.5 4.8 7.7 6.9 5.6 5 5.9 5.4 5.6 5.5 N.D. 6.5 6.2 5.2 5.3 GDP 40 38 43 70 41 36 46 48 47 40 61 56 45 44 48 50 51 57 40 62 Lactic acid 3296 3911 2249 6043 3569 3040 7913 7285 9870 8270 7878 7775 10209 5721 7800 5830 12978 6590 3993 7781 ADP 269 250 328 532 379 212 419 292 375 480 363 555 404 535 472 384 425 453 288 467 GTP 18 14 21 65 20 14 30 22 33 32 39 33 24 43 31 39 35 47 24 44 Glyoxylic acid N.D. N.D. N.D. N.D. N.D. N.D. N.D. N.D. N.D. N.D. N.D. N.D. N.D. N.D. N.D. N.D. N.D. N.D. N.D. N.D. ATP 80 55 101 382 136 67 199 95 204 336 176 234 135 422 233 226 233 303 128 259 Glycerol 3-phosphate 239 287 233 296 213 280 533 275 533 407 654 429 537 425 621 371 995 465 387 550 Glycolic acid 36 30 16 18 16 13 21 25 42 18 32 23 29 27 20 15 29 23 25 N.D. Pyruvic acid 63 66 48 117 54 92 91 129 97 155 57 34 58 68 75 135 122 129 65 109 N-Acetylglutamic acid 14 12 12 13 30 16 35 19 35 39 15 9.8 18 5.8 14 15 22 13 11 18 2-Hydroxyglutaric acid 25 29 25 15 32 21 22 24 25 27 11 35 13 18 30 9.8 26 10 10 11 Carbamoylphosphate N.D. N.D. N.D. N.D. N.D. N.D. N.D. N.D. N.D. N.D. N.D. N.D. N.D. N.D. N.D. N.D. N.D. N.D. N.D. N.D. Succinic acid 934 1167 943 361 1320 927 1385 1022 1214 1070 770 1885 1787 733 1079 292 1139 319 461 401 Malic acid 358 408 252 117 539 374 402 322 495 435 349 899 923 297 358 119 491 180 217 160 2-Oxoglutaric acid N.D. N.D. N.D. N.D. N.D. N.D. N.D. N.D. N.D. N.D. N.D. N.D. N.D. N.D. N.D. N.D. N.D. N.D. N.D. N.D. Fumaric acid 171 193 129 82 246 166 202 159 229 200 177 399 407 164 183 78 221 101 123 97 Citric acid 352 315 329 180 375 285 192 247 240 230 244 371 257 299 262 219 217 208 212 253 cis-Aconitic acid 6.3 5 5 3.5 5.7 5.5 3 4.3 4.8 4.5 4.6 6.1 4.1 5.1 4 4.8 4.1 4.5 4.1 6 Isocitric acid N.D. N.D. N.D. N.D. N.D. N.D. N.D. N.D. N.D. N.D. N.D. N.D. N.D. N.D. N.D. N.D. N.D. N.D. N.D. N.D. MS_METABOLITE_DATA_END #METABOLITES METABOLITES_START metabolite_name NAD+ cAMP cGMP NADH Xanthine ADP-ribose Mevalonic acid UDP-glucose Uric acid NADP+ IMP Sedoheptulose 7-phosphate Glucose 6-phosphate Fructose 6-phosphate Fructose 1-phosphate Galactose 1-phosphate Glucose 1-phosphate Acetoacetyl CoA Acetyl CoA Folic acid Ribose 5-phosphate CoA Ribose 1-phosphate Ribulose 5-phosphate Xylulose 5-phosphate Erythrose 4-phosphate HMG CoA Glyceraldehyde 3-phosphate NADPH Malonyl CoA Phosphocreatine XMP Dihydroxyacetone phosphate Adenylosuccinic acid Fructose 1,6-diphosphate 6-Phosphogluconic acid N-Carbamoylaspartic acid PRPP 2-Phosphoglyceric acid 2,3-Diphosphoglyceric acid 3-Phosphoglyceric acid Phosphoenolpyruvic acid GMP AMP 2-Oxoisovaleric acid GDP Lactic acid ADP GTP Glyoxylic acid ATP Glycerol 3-phosphate Glycolic acid Pyruvic acid N-Acetylglutamic acid 2-Hydroxyglutaric acid Carbamoylphosphate Succinic acid Malic acid 2-Oxoglutaric acid Fumaric acid Citric acid cis-Aconitic acid Isocitric acid METABOLITES_END #END