#METABOLOMICS WORKBENCH joerghofmann_20220406_110456 DATATRACK_ID:3177 STUDY_ID:ST002140 ANALYSIS_ID:AN003501 PROJECT_ID:PR001355
VERSION 1
CREATED_ON April 15, 2022, 1:34 pm
#PROJECT
PR:PROJECT_TITLE Mitochondrial respiration in B lymphocytes is essential for humoral immunity by
PR:PROJECT_TITLE controlling flux of the TCA cycle
PR:PROJECT_SUMMARY The function of mitochondrial respiration during B cell fate decisions and
PR:PROJECT_SUMMARY differentiation 55 remained equivocal. This study reveals that selection for
PR:PROJECT_SUMMARY mitochondrial fitness occurs during B 56 cell activation and is essential for
PR:PROJECT_SUMMARY subsequent plasma cell differentiation. By expressing a 57 mutated mitochondrial
PR:PROJECT_SUMMARY helicase in transitional B cells, we depleted mitochondrial DNA during 58 B cell
PR:PROJECT_SUMMARY maturation, resulting in reduced oxidative phosphorylation. Although no changes
PR:PROJECT_SUMMARY in 59 follicular B cell development were evident, germinal centers, class switch
PR:PROJECT_SUMMARY recombination to 60 IgG, plasma cell maturation and humoral immunity were
PR:PROJECT_SUMMARY diminished. Defective oxidative 61 phosphorylation led to aberrant flux of the
PR:PROJECT_SUMMARY tricarboxylic acid cycle and lowered the amount of 62 saturated phosphatidic
PR:PROJECT_SUMMARY acid. Consequently, mTOR activity and BLIMP1 induction were 63 curtailed whereas
PR:PROJECT_SUMMARY HIF1 _and glycolysis were amplified. Exogenous phosphatidic acid 64 increased
PR:PROJECT_SUMMARY mTOR activity in activated B cells. Hence, mitochondrial function is required
PR:PROJECT_SUMMARY and 65 selected for in activated B cells for the successful generation of
PR:PROJECT_SUMMARY functional plasma cells.
PR:INSTITUTE University of Erlangen-Nuremberg Chair of Biochemistry
PR:LAST_NAME Hofmann
PR:FIRST_NAME Joerg
PR:ADDRESS Staudtstr.5, Erlangen, Bavaria, 91058, Germany
PR:EMAIL joerg.hofmann@fau.de
PR:PHONE +49 9131 85 8260
#STUDY
ST:STUDY_TITLE Mitochondrial respiration in B lymphocytes is essential for humoral immunity by
ST:STUDY_TITLE controlling flux of the TCA cycle
ST:STUDY_SUMMARY The function of mitochondrial respiration during B cell fate decisions and
ST:STUDY_SUMMARY differentiation 55 remained equivocal. This study reveals that selection for
ST:STUDY_SUMMARY mitochondrial fitness occurs during B 56 cell activation and is essential for
ST:STUDY_SUMMARY subsequent plasma cell differentiation. By expressing a 57 mutated mitochondrial
ST:STUDY_SUMMARY helicase in transitional B cells, we depleted mitochondrial DNA during 58 B cell
ST:STUDY_SUMMARY maturation, resulting in reduced oxidative phosphorylation. Although no changes
ST:STUDY_SUMMARY in 59 follicular B cell development were evident, germinal centers, class switch
ST:STUDY_SUMMARY recombination to 60 IgG, plasma cell maturation and humoral immunity were
ST:STUDY_SUMMARY diminished. Defective oxidative 61 phosphorylation led to aberrant flux of the
ST:STUDY_SUMMARY tricarboxylic acid cycle and lowered the amount of 62 saturated phosphatidic
ST:STUDY_SUMMARY acid. Consequently, mTOR activity and BLIMP1 induction were 63 curtailed whereas
ST:STUDY_SUMMARY HIF1 _and glycolysis were amplified. Exogenous phosphatidic acid 64 increased
ST:STUDY_SUMMARY mTOR activity in activated B cells. Hence, mitochondrial function is required
ST:STUDY_SUMMARY and 65 selected for in activated B cells for the successful generation of
ST:STUDY_SUMMARY functional plasma cells.
ST:INSTITUTE University of Erlangen-Nürnberg
ST:DEPARTMENT Division of Molecular Immunology.Universitätsklinikum Erlangen, Nikolaus
ST:DEPARTMENT Fibinger Zentrum
ST:LABORATORY Prof. Mielenz
ST:LAST_NAME Mielenz
ST:FIRST_NAME Dirk
ST:ADDRESS Nikolaus-Fiebiger-Zentrum, Glückstraße 6, 91054 Erlangen
ST:EMAIL dirk.mielenz@fau.de
ST:PHONE ++49 9131 8539105
#SUBJECT
SU:SUBJECT_TYPE Cultured cells
SU:SUBJECT_SPECIES Mus musculus
SU:TAXONOMY_ID 10090
SU:GENDER Male and female
#FACTORS
#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 - 1aCre Genotype:wildtype Treatment=Crecontrol; RAW_FILE_NAME=1aCre_191204_PMet_BZellen.mzML
SUBJECT_SAMPLE_FACTORS - 2aCre Genotype:wildtype Treatment=Crecontrol; RAW_FILE_NAME=2aCre_191204_PMet_BZellen.mzML
SUBJECT_SAMPLE_FACTORS - 3aDNT Genotype:mutant Treatment=DNTwinkle; RAW_FILE_NAME=3aDNT_191204_PMet_BZellen.mzML
SUBJECT_SAMPLE_FACTORS - 4aDNT Genotype:mutant Treatment=DNTwinkle; RAW_FILE_NAME=4aDNT_191204_PMet_BZellen.mzML
SUBJECT_SAMPLE_FACTORS - 1bCre Genotype:wildtype Treatment=Crecontrol; RAW_FILE_NAME=1bCre_200312_PMet_BZellen.mzML
SUBJECT_SAMPLE_FACTORS - 2bCre Genotype:wildtype Treatment=Crecontrol; RAW_FILE_NAME=2bCre_200312_PMet_BZellen.mzML
SUBJECT_SAMPLE_FACTORS - 3bDNT Genotype:mutant Treatment=DNTwinkle; RAW_FILE_NAME=3bDNT_200312_PMet_BZellen.mzML
SUBJECT_SAMPLE_FACTORS - 4bDNT Genotype:mutant Treatment=DNTwinkle; RAW_FILE_NAME=4bDNT_200312_PMet_BZellen.mzML
SUBJECT_SAMPLE_FACTORS - 1cCre Genotype:wildtype Treatment=Crecontrol; RAW_FILE_NAME=1cCre_210706_PMet_BZellen.mzML
SUBJECT_SAMPLE_FACTORS - 2cCre Genotype:wildtype Treatment=Crecontrol; RAW_FILE_NAME=2cCre_210706_PMet_BZellen.mzML
SUBJECT_SAMPLE_FACTORS - 3cCre Genotype:wildtype Treatment=Crecontrol; RAW_FILE_NAME=3cCre_210706_PMet_BZellen.mzML
SUBJECT_SAMPLE_FACTORS - 4cDNT Genotype:mutant Treatment=DNTwinkle; RAW_FILE_NAME=4cDNT_210706_PMet_BZellen.mzML
SUBJECT_SAMPLE_FACTORS - 5cDNT Genotype:mutant Treatment=DNTwinkle; RAW_FILE_NAME=5cDNT_210706_PMet_BZellen.mzML
SUBJECT_SAMPLE_FACTORS - 6cDNT Genotype:mutant Treatment=DNTwinkle; RAW_FILE_NAME=6cDNT_210706_PMet_BZellen.mzML
SUBJECT_SAMPLE_FACTORS - B cells_DNT_GPL analysis 1-S01_DNT Genotype:mutant Treatment=DNTwinkle; RAW_FILE_NAME=B cells_DNT_GPL analysis 1-S01_DNT.mzML
SUBJECT_SAMPLE_FACTORS - B cells_DNT_GPL analysis 1-S02_Cre Genotype:wildtype Treatment=Crecontrol; RAW_FILE_NAME=B cells_DNT_GPL analysis 1-S02_Cre.mzML
SUBJECT_SAMPLE_FACTORS - B cells_DNT_GPL analysis 1-S03_DNT Genotype:mutant Treatment=DNTwinkle; RAW_FILE_NAME=B cells_DNT_GPL analysis 1-S03_DNT.mzML
SUBJECT_SAMPLE_FACTORS - B cells_DNT_GPL analysis 1-S04_Cre Genotype:wildtype Treatment=Crecontrol; RAW_FILE_NAME=B cells_DNT_GPL analysis 1-S04_Cre.mzML
SUBJECT_SAMPLE_FACTORS - B cells_DNT_GPL analysis 1-S05_DNT Genotype:mutant Treatment=DNTwinkle; RAW_FILE_NAME=B cells_DNT_GPL analysis 1-S05_DNT.mzML
SUBJECT_SAMPLE_FACTORS - B cells_DNT_GPL analysis 1-S06_Cre Genotype:wildtype Treatment=Crecontrol; RAW_FILE_NAME=B cells_DNT_GPL analysis 1-S06_Cre.mzML
SUBJECT_SAMPLE_FACTORS - B cells_DNT_GPL analysis 2b-S01_Cre1 Genotype:wildtype Treatment=Crecontrol; RAW_FILE_NAME=B cells_DNT_GPL analysis 2b-S01_Cre1.mzML
SUBJECT_SAMPLE_FACTORS - B cells_DNT_GPL analysis 2b-S02_DNT2 Genotype:mutant Treatment=DNTwinkle; RAW_FILE_NAME=B cells_DNT_GPL analysis 2b-S02_DNT2.mzML
SUBJECT_SAMPLE_FACTORS - B cells_DNT_GPL analysis 2b-S03_Cre3 Genotype:wildtype Treatment=Crecontrol; RAW_FILE_NAME=B cells_DNT_GPL analysis 2b-S03_Cre3.mzML
SUBJECT_SAMPLE_FACTORS - B cells_DNT_GPL analysis 2b-S04_DNT4 Genotype:mutant Treatment=DNTwinkle; RAW_FILE_NAME=B cells_DNT_GPL analysis 2b-S04_DNT4.mzML
SUBJECT_SAMPLE_FACTORS - B cells_DNT_GPL analysis 2b-S05_Cre5 Genotype:wildtype Treatment=Crecontrol; RAW_FILE_NAME=B cells_DNT_GPL analysis 2b-S05_Cre5.mzML
SUBJECT_SAMPLE_FACTORS - B cells_DNT_GPL analysis 2b-S06_DNT6 Genotype:mutant Treatment=DNTwinkle; RAW_FILE_NAME=B cells_DNT_GPL analysis 2b-S06_DNT6.mzML
SUBJECT_SAMPLE_FACTORS - B cells_DNT_GPL analysis 2b-S07_Cre7 Genotype:wildtype Treatment=Crecontrol; RAW_FILE_NAME=B cells_DNT_GPL analysis 2b-S07_Cre7.mzML
SUBJECT_SAMPLE_FACTORS - B cells_DNT_GPL analysis 2b-S08_DNT8 Genotype:mutant Treatment=DNTwinkle; RAW_FILE_NAME=B cells_DNT_GPL analysis 2b-S08_DNT8.mzML
SUBJECT_SAMPLE_FACTORS - B cells_DNT_GPL analysis 2b-S10_DNT10 Genotype:mutant Treatment=DNTwinkle; RAW_FILE_NAME=B cells_DNT_GPL analysis 2b-S10_DNT10.mzML
#COLLECTION
CO:COLLECTION_SUMMARY Both female and male mice were used in the experiments. Mice were maintained on
CO:COLLECTION_SUMMARY a 12-h light/dark cycle with free access to food and water according to
CO:COLLECTION_SUMMARY governmental rules. K320E-TWINKLE floxed mice (Baris et al., 2015) were crossed
CO:COLLECTION_SUMMARY to CD23 CRE mice (Kwon et al., 2008) kindly provided by Meinrad Busslinger) to
CO:COLLECTION_SUMMARY generate DNT animals. DNT mice used in these experiments had the genetic
CO:COLLECTION_SUMMARY background DNT+/- CRE+/- and CRE control mice were DNT-/- CRE+/-. The WT animals
CO:COLLECTION_SUMMARY used in this study were DNT-/- CRE-/- littermates. All mice were on the C57Bl/6
CO:COLLECTION_SUMMARY background. Isolation of primary murine cells from spleen and bone marrow Spleen
CO:COLLECTION_SUMMARY was transferred into cold 2 % FCS (in PBS) and gently passed through a 70 µm
CO:COLLECTION_SUMMARY cell strainer (BD) using the plunger of a 5 ml syringe (BD). Femur and tibia
CO:COLLECTION_SUMMARY were flushed with cold 2 % FCS using a 27 G cannula (BD). Cell suspensions were
CO:COLLECTION_SUMMARY pelleted by centrifugation at 300 x g for 5 min at 4°C. Erythrocytes were lysed
CO:COLLECTION_SUMMARY in red blood cell-lysis buffer (150 mM NH4Cl, 10 mM KHCO3, 100 µM EDTA) for
CO:COLLECTION_SUMMARY 5min at room temperature. The reaction was stopped by adding cold 2% FCS before
CO:COLLECTION_SUMMARY centrifugation at 300 x g for 5 min at 4°C. The final cell suspensions were
CO:COLLECTION_SUMMARY kept in cold 2 % FCS after filtration through 30 µm mesh filter (Sysmex). In
CO:COLLECTION_SUMMARY vitro cultivation of primary murine B cells Splenic B cells were cultured with a
CO:COLLECTION_SUMMARY starting concentration of 0.5 x 106 cells/ ml in R10 medium (RPMI1640, 10 %
CO:COLLECTION_SUMMARY fetal calf serum (FCS), 2 mM glutamate, 1 mM sodium pyruvate, 50 U/ml penicillin
CO:COLLECTION_SUMMARY G, 50 μg/ml streptomycin, 50 μM β-mercaptoethanol) for 72 h at 37°C and 5%
CO:COLLECTION_SUMMARY CO2, supplemented with 10 µg/ml LPS. For in vitro class switch recombination
CO:COLLECTION_SUMMARY cells were seeded at 0.1 x 106 cells/ ml in R10 medium for 96 h, supplemented
CO:COLLECTION_SUMMARY with 5 ng/ml transforming growth factor , 5 nM retinoic acid, 10 µg/ml
CO:COLLECTION_SUMMARY anti-CD40 antibody, 10 µg/ml LPS, 100 U/ml IL4 and 10 ng/ml IL5. Ref.: Baris,
CO:COLLECTION_SUMMARY O.R., Ederer, S., Neuhaus, J.F., von Kleist-Retzow, J.C., Wunderlich, C.M., Pal,
CO:COLLECTION_SUMMARY M., WunderlichF.T., Peeva, V., Zsurka, G., Kunz, W.S., et al. (2015). Mosaic
CO:COLLECTION_SUMMARY Deficiency in Mitochondrial Oxidative Metabolism Promotes Cardiac Arrhythmia
CO:COLLECTION_SUMMARY during Aging. Cell Metab 21, 667–677.
CO:COLLECTION_PROTOCOL_FILENAME Bcellscoll Mielenz.pdf
CO:SAMPLE_TYPE B-cells
#TREATMENT
TR:TREATMENT_SUMMARY in vitro activated (LPS) B cells, which cannot replicate their mitochondrial DNA
TR:TREATMENT_SUMMARY ("DNTwinkle") resulting in impaired respiratory chain activity and oxidative
TR:TREATMENT_SUMMARY phosphorylation compared to Cre control B cells DTN Dominant Negative Twinkle
TR:TREATMENT_SUMMARY (mitochondtail Helicase)
#SAMPLEPREP
SP:SAMPLEPREP_SUMMARY Glycerophospholipid analysis Glycerophospholipids (PCH, PE, PI, PS, PG, PA) in B
SP:SAMPLEPREP_SUMMARY cells were analyzed by Nano- Electrospray Ionization Tandem Mass Spectrometry
SP:SAMPLEPREP_SUMMARY (Nano-ESI-MS/MS) with direct infusion of the lipid extract (Shotgun Lipidomics):
SP:SAMPLEPREP_SUMMARY 14 to 45 x 106 cells were homogenized in 300 μl of Milli- Q water using the
SP:SAMPLEPREP_SUMMARY Precellys 24 Homogenisator (Peqlab, Erlangen, Germany) at 6.500 rpm for 30 sec.
SP:SAMPLEPREP_SUMMARY The protein content of the homogenate was routinely determined using
SP:SAMPLEPREP_SUMMARY bicinchoninic acid. To 100 μl of the homogenate 400 μl of Milli-Q water,
SP:SAMPLEPREP_SUMMARY 1.875ml of methanol/chloroform 2:1 (v/v) and internal standards (125 pmol PCH
SP:SAMPLEPREP_SUMMARY 17:0-20:4, 132 pmol PE 17:0-20:4, 118 pmol PI 17:0-20:4, 131 pmol PS 17:0-20:4,
SP:SAMPLEPREP_SUMMARY 62 pmol PG 17:0/20:4, 75 pmol PA 17:0/20:4 Avanti Polar Lipids) were added.
SP:SAMPLEPREP_SUMMARY Lipid extraction and Nano-ESI-MS/MS analysis were performed as previously
SP:SAMPLEPREP_SUMMARY described (Kumar et al., 2015). Endogenous glycerophospolipids were quantified
SP:SAMPLEPREP_SUMMARY by referring their peak areas to those of the internal standards. The calculated
SP:SAMPLEPREP_SUMMARY glycerophospolipid amounts were normalized to the protein content of the tissue
SP:SAMPLEPREP_SUMMARY homogenate. Metabolomics of phosphorylated metabolites und carbonic acids
SP:SAMPLEPREP_SUMMARY Experimental Setup I: Splenic B cells were isolated, activated with LPS and
SP:SAMPLEPREP_SUMMARY viable cells, only GFP+ for DNT, were sorted after 3 days using flow cytometry.
SP:SAMPLEPREP_SUMMARY Perchloric acid extraction and metabolic profiling was performed as previously
SP:SAMPLEPREP_SUMMARY published measured by LCMS/MS on an QTrap 3200 (Sciex) (Hofmann et al., 2011).
SP:SAMPLEPREP_SUMMARY Ref.: Kumar, V., Bouameur, J.E., Bar, J., Rice, R.H., Hornig-Do, H.T., Roop,
SP:SAMPLEPREP_SUMMARY D.R., Schwarz, N., Brodesser, 1120 S., Thiering, S., Leube, R.E., et al. (2015).
SP:SAMPLEPREP_SUMMARY A keratin scaffold regulates epidermal barrier 1121 formation, mitochondrial
SP:SAMPLEPREP_SUMMARY lipid composition, and activity. J Cell Biol 211, 1057–1075. Ref.: Hofmann,
SP:SAMPLEPREP_SUMMARY J., Bornke, F., Schmiedl, A., Kleine, T., and Sonnewald, U. (2011). Detecting
SP:SAMPLEPREP_SUMMARY functional groups of Arabidopsis mutants by metabolic profiling and evaluation
SP:SAMPLEPREP_SUMMARY of pleiotropic responses. 10Front Plant Sci 2, 82.
#CHROMATOGRAPHY
CH:CHROMATOGRAPHY_SUMMARY The HPLC-system was controlled by the software Chromeleon VS 6.8 and DCMS-Link
CH:CHROMATOGRAPHY_SUMMARY VS1.1 (Dionex) in combination with Analyst 1.4.1 (Applied Biosystems).
CH:CHROMATOGRAPHY_SUMMARY Metabolites were separated on two IonPac AS11HC columns (2 × 250 mm; Dionex)
CH:CHROMATOGRAPHY_SUMMARY protected by an AG11HC guard column (2 × 50 mm). The elution gradient was
CH:CHROMATOGRAPHY_SUMMARY generated with water (eluent A) and 100 mm KOH (eluent B) within a total run
CH:CHROMATOGRAPHY_SUMMARY time of 80 min at a flow rate of 0.25 mL min−1 and a column temperature of
CH:CHROMATOGRAPHY_SUMMARY 35°C as follows: 0 min, 4%; 0 to 1 min, 4%; 1 to 6 min, 15%; 6 to 12 min, 19%;
CH:CHROMATOGRAPHY_SUMMARY 12 to 22 min, 20%; 22 to 24 min, 23%; 24 to 27 min, 35%; 27 to 37 min, 38%; 37
CH:CHROMATOGRAPHY_SUMMARY to 39 min, 45%; 39 to 44 min, 100%; 44 to 71 min, 100%; 71 to 76 min, 4%; and 76
CH:CHROMATOGRAPHY_SUMMARY to 80 min, 4% eluent B. Ref.: Hofmann, J., Bornke, F., Schmiedl, A., Kleine, T.,
CH:CHROMATOGRAPHY_SUMMARY and Sonnewald, U. (2011). Detecting functional groups of Arabidopsis mutants by
CH:CHROMATOGRAPHY_SUMMARY metabolic profiling and evaluation of pleiotropic responses. 10Front Plant Sci
CH:CHROMATOGRAPHY_SUMMARY 2, 82.
CH:CHROMATOGRAPHY_TYPE Ion Chromatography
CH:INSTRUMENT_NAME ThermoDionexICS3000
CH:COLUMN_NAME ThermoDionexAS11/AG11
#ANALYSIS
AN:ANALYSIS_TYPE MS
#MS
MS:INSTRUMENT_NAME ABI Sciex 3200 QTrap
MS:INSTRUMENT_TYPE QTRAP
MS:MS_TYPE ESI
MS:ION_MODE NEGATIVE
MS:MS_COMMENTS Mass spectrometric analysis was performed using the QTRAP 3200 (SCIEX) operated
MS:MS_COMMENTS by Analyst 1.6.2. The electrospray ionization source parameters were −4,500 eV
MS:MS_COMMENTS at 600°C, N2 gas pressures were 20 p.s.i. (curtaingas), 30 p.s.i. (gas1), and
MS:MS_COMMENTS 20 p.s.i. (gas2), and collision gas was set to medium. The dwell time for ions
MS:MS_COMMENTS was 75 ms, and scan time per cycle was 3.7 s.Scan ranges were from
MS:MS_COMMENTS mass-to-charge ratio 87 to 606 (precursor ions) and mass-to-charge ratio 59 to
MS:MS_COMMENTS 385 (product ions). Masstransitions (metabolite m/z mother ion / m/z daughter
MS:MS_COMMENTS ion)recorded were as follows: UDP-glucose 565 / 323, glucose-1-phosphate 259 /
MS:MS_COMMENTS 79, glucose-6-phosphate 259 / 97, 3-phosphoglycerate 185 / 97, phosphoenol
MS:MS_COMMENTS pyruvate 167 / 79, citrate 191 / 87, isocitrate 191 / 111, malate 133 / 71, AMP
MS:MS_COMMENTS 346/79, ADP 427/79, 2-oxoglutarate (aKG) 145 / 101, succinate 117 / 73, UDPNAG
MS:MS_COMMENTS 606/385, Itaconate 129/85, Lactat 89/43, D-glucose 179/89, fumarate 115 / 71,
MS:MS_COMMENTS E4P 199/97, ATP 427/79, UDP 404/79, G16BP 339/97 The contents of metabolites
MS:MS_COMMENTS were calculated based on peak areas for precursor/product ion transitions
MS:MS_COMMENTS relative to standards.
#MS_METABOLITE_DATA
MS_METABOLITE_DATA:UNITS pmol/10E6 cells
MS_METABOLITE_DATA_START
Samples 1aCre 2aCre 3aDNT 4aDNT 1bCre 2bCre 3bDNT 4bDNT 1cCre 2cCre 3cCre 4cDNT 5cDNT 5cDNT
Factors Genotype:wildtype Genotype:wildtype Genotype:mutant Genotype:mutant Genotype:wildtype Genotype:wildtype Genotype:mutant Genotype:mutant Genotype:wildtype Genotype:wildtype Genotype:wildtype Genotype:mutant Genotype:mutant Genotype:mutant
G1P 5 4 3 3 2 3
G6P 0 0 0 1 12 10 8 10 8 9
3PG 1 1 1 1 8 8 3 3 40 23 22 10 9 10
PEP 0 1 0 0 9 7 3 1 20 13 13 8 5 4
Cit 6 8 5 5 47 48 12 10 256 234 160 111 62 60
Icit 1 0 0 0 2 1 0 0 20 11 11 3 5 5
Mal 59 56 187 183 177 136 457 391 511 393 466 902 856 901
AMP 9 8 10 16 45 13 40 17 128 83 152 114 60 116
ADP 18 18 21 14 82 55 79 56 394 564 392 231 174 219
aKG 4 2 2 3 5 2 4 2 37 29 28 61 109 67
Succ 45 38 67 128 159 85 57 97 329 293 255 299 294 290
UDPNAG 2 3 1 1 4 4 2 1 139 123 90 39 34 25
Itaconat 3 1 1 1 3 1 0 1 25 23 21 26 42 23
Lactat 46 4 10 34 13 8 47 62 140 226 142 265 445 249
D-Glucose 91 180 69 317 157 162 274 104 28 129 37 388 794 349
Fum 15 10 30 36 13 11 50 41 132 107 149 291 219 310
E4P 0 0 0 0 2 2 5 4 96 78 63 78 112 107
ATP 4 6 3 3 112 217 76 97 2187 2650 1539 1255 1458 1197
UDP 9 10 9 4 217 250 162 452 68 69
G16BP 10 13 6 4 6 9
MS_METABOLITE_DATA_END
#METABOLITES
METABOLITES_START
metabolite_name Transition measured CAS ID
G1P 259_97 59-56-3
G6P 259_79 56-73-5
3PG 185_97 820-11-1
PEP 167_79 138-08-9
Cit 191_87 77-92-9
Icit 191_111 320-77-4
Mal 133_115 636-61-3
AMP 346_79 61-19-8
ADP 427_79 58-64-0
aKG 145_101 328-50-7
Succ 117_73 110-15-6
UDPNAG 606_385 91183-98-1
Itaconat 129_85 97-65-4
Lactat 89_43 113-21-3
D-Glucose 179_ 89 50-99-7
Fum 115_71 110-17-8
E4P 199_97 585-18-2
ATP 427_79 56-65-5
UDP 404_79 58-98-0
G16BP 339_97 305-58-8
METABOLITES_END
#END