#METABOLOMICS WORKBENCH Brodesser_20230816_054636 DATATRACK_ID:4224 STUDY_ID:ST003114 ANALYSIS_ID:AN005102 PROJECT_ID:PR001935 VERSION 1 CREATED_ON March 6, 2024, 10:51 am #PROJECT PR:PROJECT_TITLE Lipid unsaturation promotes BAX and BAK pore activity during apoptosis PR:PROJECT_SUMMARY BAX and BAK are proapoptotic members of the BCL2 family that directly mediate PR:PROJECT_SUMMARY mitochondrial outer membrane permeabilization (MOMP), a central step in PR:PROJECT_SUMMARY apoptosis execution. However, the molecular architecture of the mitochondrial PR:PROJECT_SUMMARY apoptotic pore remains a key open question and especially little is known about PR:PROJECT_SUMMARY the contribution of lipids to MOMP. By performing a comparative lipidomics PR:PROJECT_SUMMARY analysis of the proximal membrane environment of BAK isolated in lipid PR:PROJECT_SUMMARY nanodiscs, we find a significant enrichment of unsaturated species nearby BAK PR:PROJECT_SUMMARY and BAX in apoptotic conditions. We then demonstrate that unsaturated lipids PR:PROJECT_SUMMARY promote BAX pore activity in model membranes, isolated mitochondria and cellular PR:PROJECT_SUMMARY systems, which is further supported by molecular dynamics simulations. PR:PROJECT_SUMMARY Accordingly, the fatty acid desaturase FADS2 not only enhances apoptosis PR:PROJECT_SUMMARY sensitivity, but also the activation of the cGAS/STING pathway downstream mtDNA PR:PROJECT_SUMMARY release. The correlation of FADS2 levels with the sensitization to apoptosis of PR:PROJECT_SUMMARY different lung and kidney cancer cell lines by co-treatment with unsaturated PR:PROJECT_SUMMARY fatty acids supports the relevance of our findings. Altogether, our work PR:PROJECT_SUMMARY provides new insight on how local lipid environment affects BAX and BAK function PR:PROJECT_SUMMARY during apoptosis. PR:INSTITUTE University of Cologne PR:DEPARTMENT Institute for Genetics, Cluster of Excellence Cellular Stress Responses in PR:DEPARTMENT Aging-associated Diseases (CECAD) PR:LAST_NAME García-Sáez PR:FIRST_NAME Ana J. PR:ADDRESS Joseph-Stelzmann-Str. 26, 50931 Cologne, Germany PR:EMAIL ana.garcia@uni-koeln.de PR:PHONE +49 221 478 84261 PR:CONTRIBUTORS Shashank Dadsena, Rodrigo Cuevas Arenas, Gonçalo Vieira, Susanne Brodesser, PR:CONTRIBUTORS Manuel N. Melo, Ana J. García-Sáez #STUDY ST:STUDY_TITLE Lipidomics analyses in model membranes, isolated mitochondria and cellular ST:STUDY_TITLE systems to study how the local lipid environment affects BAX and BAK function ST:STUDY_TITLE during apoptosis. ST:STUDY_SUMMARY To investigate how the local lipid environment affects BAX and BAK function ST:STUDY_SUMMARY during apoptosis, we performed quantitative analyses of different lipid classes ST:STUDY_SUMMARY (glycerophospholipids, fatty acids, ceramides and sphingomyelins) in cultured ST:STUDY_SUMMARY cells, isolated mitochondria and lipid nanodics formed by Styrene-Malic Acid ST:STUDY_SUMMARY (SMA) co-polymers. Ceramides, sphingomyelins, fatty acids and cardiolipins were ST:STUDY_SUMMARY analyzed by Liquid Chromatography coupled to Tandem Mass Spectrometry ST:STUDY_SUMMARY (LC-MS/MS). For glycerophospholipids (PC, PE, PI, PS, PG, PA) we applied direct ST:STUDY_SUMMARY infusion MS approaches (Shotgun Lipidomics). ST:INSTITUTE University of Cologne ST:DEPARTMENT Faculty of Medicine and University Hospital of Cologne, Cluster of Excellence ST:DEPARTMENT Cellular Stress Responses in Aging-associated Diseases (CECAD) ST:LABORATORY CECAD Lipidomics/Metabolomics Facility ST:LAST_NAME Brodesser ST:FIRST_NAME Susanne ST:ADDRESS Joseph-Stelzmann-Str. 26, 50931 Cologne, Germany ST:EMAIL susanne.brodesser@uk-koeln.de ST:PHONE +49 221 478 84015 #SUBJECT SU:SUBJECT_TYPE Cultured cells SU:SUBJECT_SPECIES Homo sapiens SU:TAXONOMY_ID 9606 #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 - S01_mitos_control.SMA_1 Sample source:mitochondrial SMALPs | Genotype:WT | Condition:control RAW_FILE_NAME=GPL_S01_mitos_control.SMA_1.mzML; RAW_FILE_NAME=CerSM_S01_mitos_control.SMA_1.mzML; RAW_FILE_NAME=CL_S01_mitos_control.SMA_1.mzML; RAW_FILE_NAME=- SUBJECT_SAMPLE_FACTORS - S02_mitos_control.SMA_2 Sample source:mitochondrial SMALPs | Genotype:WT | Condition:control RAW_FILE_NAME=GPL_S02_mitos_control.SMA_2.mzML; RAW_FILE_NAME=CerSM_S02_mitos_control.SMA_2.mzML; RAW_FILE_NAME=CL_S02_mitos_control.SMA_2.mzML; RAW_FILE_NAME=- SUBJECT_SAMPLE_FACTORS - S03_mitos_control.SMA_3 Sample source:mitochondrial SMALPs | Genotype:WT | Condition:control RAW_FILE_NAME=-; RAW_FILE_NAME=CerSM_S03_mitos_control.SMA_3.mzML; RAW_FILE_NAME=CL_S03_mitos_control.SMA_3.mzML; RAW_FILE_NAME=- SUBJECT_SAMPLE_FACTORS - S04_mitos_control.SMA_4 Sample source:mitochondrial SMALPs | Genotype:WT | Condition:control RAW_FILE_NAME=GPL_S04_mitos_control.SMA_4.mzML; RAW_FILE_NAME=CerSM_S04_mitos_control.SMA_4.mzML; RAW_FILE_NAME=CL_S04_mitos_control.SMA_4.mzML; RAW_FILE_NAME=- SUBJECT_SAMPLE_FACTORS - S05_mitos_apoptosis.SMA_1 Sample source:mitochondrial SMALPs | Genotype:WT | Condition:apoptotic RAW_FILE_NAME=GPL_S05_mitos_apoptosis.SMA_1.mzML; RAW_FILE_NAME=CerSM_S05_mitos_apoptosis.SMA_1.mzML; RAW_FILE_NAME=CL_S05_mitos_apoptosis.SMA_1.mzML; RAW_FILE_NAME=- SUBJECT_SAMPLE_FACTORS - S06_mitos_apoptosis.SMA_2 Sample source:mitochondrial SMALPs | Genotype:WT | Condition:apoptotic RAW_FILE_NAME=GPL_S06_mitos_apoptosis.SMA_2.mzML; RAW_FILE_NAME=CerSM_S06_mitos_apoptosis.SMA_2.mzML; RAW_FILE_NAME=CL_S06_mitos_apoptosis.SMA_2.mzML; RAW_FILE_NAME=- SUBJECT_SAMPLE_FACTORS - S07_mitos_apoptosis.SMA_3 Sample source:mitochondrial SMALPs | Genotype:WT | Condition:apoptotic RAW_FILE_NAME=GPL_S07_mitos_apoptosis.SMA_3.mzML; RAW_FILE_NAME=CerSM_S07_mitos_apoptosis.SMA_3.mzML; RAW_FILE_NAME=CL_S07_mitos_apoptosis.SMA_3.mzML; RAW_FILE_NAME=- SUBJECT_SAMPLE_FACTORS - S08_mitos_apoptosis.SMA_4 Sample source:mitochondrial SMALPs | Genotype:WT | Condition:apoptotic RAW_FILE_NAME=-; RAW_FILE_NAME=CerSM_S08_mitos_apoptosis.SMA_4.mzML; RAW_FILE_NAME=CL_S08_mitos_apoptosis.SMA_4.mzML; RAW_FILE_NAME=- SUBJECT_SAMPLE_FACTORS - S09_mitos_control_1 Sample source:total mitochondria | Genotype:WT | Condition:control RAW_FILE_NAME=GPL_S09_mitos_control_1.mzML; RAW_FILE_NAME=CerSM_S09_mitos_control_1.mzML; RAW_FILE_NAME=CL_S09_mitos_control_1.mzML; RAW_FILE_NAME=- SUBJECT_SAMPLE_FACTORS - S10_mitos_control_2 Sample source:total mitochondria | Genotype:WT | Condition:control RAW_FILE_NAME=-; RAW_FILE_NAME=CerSM_S10_mitos_control_2.mzML; RAW_FILE_NAME=CL_S10_mitos_control_2.mzML; RAW_FILE_NAME=- SUBJECT_SAMPLE_FACTORS - S11_mitos_control_3 Sample source:total mitochondria | Genotype:WT | Condition:control RAW_FILE_NAME=GPL_S11_mitos_control_3.mzML; RAW_FILE_NAME=CerSM_S11_mitos_control_3.mzML; RAW_FILE_NAME=CL_S11_mitos_control_3.mzML; RAW_FILE_NAME=- SUBJECT_SAMPLE_FACTORS - S12_mitos_control_4 Sample source:total mitochondria | Genotype:WT | Condition:control RAW_FILE_NAME=GPL_S12_mitos_control_4.mzML; RAW_FILE_NAME=CerSM_S12_mitos_control_4.mzML; RAW_FILE_NAME=CL_S12_mitos_control_4.mzML; RAW_FILE_NAME=- SUBJECT_SAMPLE_FACTORS - S13_mitos_apoptosis_1 Sample source:total mitochondria | Genotype:WT | Condition:apoptotic RAW_FILE_NAME=GPL_S13_mitos_apoptosis_1.mzML; RAW_FILE_NAME=CerSM_S13_mitos_apoptosis_1.mzML; RAW_FILE_NAME=CL_S13_mitos_apoptosis_1.mzML; RAW_FILE_NAME=- SUBJECT_SAMPLE_FACTORS - S14_mitos_apoptosis_2 Sample source:total mitochondria | Genotype:WT | Condition:apoptotic RAW_FILE_NAME=-; RAW_FILE_NAME=CerSM_S14_mitos_apoptosis_2.mzML; RAW_FILE_NAME=CL_S14_mitos_apoptosis_2.mzML; RAW_FILE_NAME=- SUBJECT_SAMPLE_FACTORS - S15_mitos_apoptosis_3 Sample source:total mitochondria | Genotype:WT | Condition:apoptotic RAW_FILE_NAME=GPL_S15_mitos_apoptosis_3.mzML; RAW_FILE_NAME=CerSM_S15_mitos_apoptosis_3.mzML; RAW_FILE_NAME=CL_S15_mitos_apoptosis_3.mzML; RAW_FILE_NAME=- SUBJECT_SAMPLE_FACTORS - S16_mitos_apoptosis_4 Sample source:total mitochondria | Genotype:WT | Condition:apoptotic RAW_FILE_NAME=GPL_S16_mitos_apoptosis_4.mzML; RAW_FILE_NAME=CerSM_S16_mitos_apoptosis_4.mzML; RAW_FILE_NAME=CL_S16_mitos_apoptosis_4.mzML; RAW_FILE_NAME=- SUBJECT_SAMPLE_FACTORS - S02_pulldown_Control_22.08 Sample source:mitochondrial SMALPs | Genotype:mEGFP-BAK | Condition:control RAW_FILE_NAME=GPL_S02_pulldown_Control_22.08.mzML; RAW_FILE_NAME=CerSM_S02_pulldown_Control_22.08.mzML; RAW_FILE_NAME=-; RAW_FILE_NAME=- SUBJECT_SAMPLE_FACTORS - S03_pulldown_Control_01.09 Sample source:mitochondrial SMALPs | Genotype:mEGFP-BAK | Condition:control RAW_FILE_NAME=GPL_S03_pulldown_Control_01.09.mzML; RAW_FILE_NAME=CerSM_S03_pulldown_Control_01.09.mzML; RAW_FILE_NAME=-; RAW_FILE_NAME=- SUBJECT_SAMPLE_FACTORS - S04_pulldown_Control_04.09 Sample source:mitochondrial SMALPs | Genotype:mEGFP-BAK | Condition:control RAW_FILE_NAME=GPL_S04_pulldown_Control_04.09.mzML; RAW_FILE_NAME=CerSM_S04_pulldown_Control_04.09.mzML; RAW_FILE_NAME=-; RAW_FILE_NAME=- SUBJECT_SAMPLE_FACTORS - S05_pulldown_Control_31.09 Sample source:mitochondrial SMALPs | Genotype:mEGFP-BAK | Condition:control RAW_FILE_NAME=GPL_S05_pulldown_Control_31.09.mzML; RAW_FILE_NAME=CerSM_S05_pulldown_Control_31.09.mzML; RAW_FILE_NAME=-; RAW_FILE_NAME=- SUBJECT_SAMPLE_FACTORS - S06_pulldown_Apoptosis_22.08 Sample source:mitochondrial SMALPs | Genotype:mEGFP-BAK | Condition:apoptotic RAW_FILE_NAME=GPL_S06_pulldown_Apoptosis_22.08.mzML; RAW_FILE_NAME=CerSM_S06_pulldown_Apoptosis_22.08.mzML; RAW_FILE_NAME=-; RAW_FILE_NAME=- SUBJECT_SAMPLE_FACTORS - S07_pulldown_Apoptosis_01.09 Sample source:mitochondrial SMALPs | Genotype:mEGFP-BAK | Condition:apoptotic RAW_FILE_NAME=GPL_S07_pulldown_Apoptosis_01.09.mzML; RAW_FILE_NAME=CerSM_S07_pulldown_Apoptosis_01.09.mzML; RAW_FILE_NAME=-; RAW_FILE_NAME=- SUBJECT_SAMPLE_FACTORS - S08_pulldown_Apoptosis_04.09 Sample source:mitochondrial SMALPs | Genotype:mEGFP-BAK | Condition:apoptotic RAW_FILE_NAME=GPL_S08_pulldown_Apoptosis_04.09.mzML; RAW_FILE_NAME=CerSM_S08_pulldown_Apoptosis_04.09.mzML; RAW_FILE_NAME=-; RAW_FILE_NAME=- SUBJECT_SAMPLE_FACTORS - S09_pulldown_Apoptosis_31.09 Sample source:mitochondrial SMALPs | Genotype:mEGFP-BAK | Condition:apoptotic RAW_FILE_NAME=GPL_S09_pulldown_Apoptosis_31.09.mzML; RAW_FILE_NAME=CerSM_S09_pulldown_Apoptosis_31.09.mzML; RAW_FILE_NAME=-; RAW_FILE_NAME=- SUBJECT_SAMPLE_FACTORS - S01_mitos_no.treatment_WT_1 Sample source:total mitochondria | Genotype:WT | Condition:untreated RAW_FILE_NAME=-; RAW_FILE_NAME=-; RAW_FILE_NAME=-; RAW_FILE_NAME=FA_S01_mitos_no.treatment_WT_1.mzML SUBJECT_SAMPLE_FACTORS - S02_mitos_no.treatment_WT_2 Sample source:total mitochondria | Genotype:WT | Condition:untreated RAW_FILE_NAME=-; RAW_FILE_NAME=-; RAW_FILE_NAME=-; RAW_FILE_NAME=FA_S02_mitos_no.treatment_WT_2.mzML SUBJECT_SAMPLE_FACTORS - S03_mitos_no.treatment_WT_3 Sample source:total mitochondria | Genotype:WT | Condition:untreated RAW_FILE_NAME=-; RAW_FILE_NAME=-; RAW_FILE_NAME=-; RAW_FILE_NAME=FA_S03_mitos_no.treatment_WT_3.mzML SUBJECT_SAMPLE_FACTORS - S04_mitos_no.treatment_WT_4 Sample source:total mitochondria | Genotype:WT | Condition:untreated RAW_FILE_NAME=-; RAW_FILE_NAME=-; RAW_FILE_NAME=-; RAW_FILE_NAME=FA_S04_mitos_no.treatment_WT_4.mzML SUBJECT_SAMPLE_FACTORS - S05_mitos_linoleic.acid_WT_1 Sample source:total mitochondria | Genotype:WT | Condition:linoleic acid RAW_FILE_NAME=-; RAW_FILE_NAME=-; RAW_FILE_NAME=-; RAW_FILE_NAME=FA_S05_mitos_linoleic.acid_WT_1.mzML SUBJECT_SAMPLE_FACTORS - S06_mitos_linoleic.acid_WT_2 Sample source:total mitochondria | Genotype:WT | Condition:linoleic acid RAW_FILE_NAME=-; RAW_FILE_NAME=-; RAW_FILE_NAME=-; RAW_FILE_NAME=FA_S06_mitos_linoleic.acid_WT_2.mzML SUBJECT_SAMPLE_FACTORS - S07_mitos_linoleic.acid_WT_3 Sample source:total mitochondria | Genotype:WT | Condition:linoleic acid RAW_FILE_NAME=-; RAW_FILE_NAME=-; RAW_FILE_NAME=-; RAW_FILE_NAME=FA_S07_mitos_linoleic.acid_WT_3.mzML SUBJECT_SAMPLE_FACTORS - S08_mitos_linoleic.acid_WT_4 Sample source:total mitochondria | Genotype:WT | Condition:linoleic acid RAW_FILE_NAME=-; RAW_FILE_NAME=-; RAW_FILE_NAME=-; RAW_FILE_NAME=FA_S08_mitos_linoleic.acid_WT_4.mzML SUBJECT_SAMPLE_FACTORS - S09_smitos_no.treatment_KO_1 Sample source:total mitochondria | Genotype:FADS2 KO | Condition:untreated RAW_FILE_NAME=-; RAW_FILE_NAME=-; RAW_FILE_NAME=-; RAW_FILE_NAME=FA_S09_mitos_no.treatment_KO_1.mzML SUBJECT_SAMPLE_FACTORS - S10_mitos_no.treatment_KO_2 Sample source:total mitochondria | Genotype:FADS2 KO | Condition:untreated RAW_FILE_NAME=-; RAW_FILE_NAME=-; RAW_FILE_NAME=-; RAW_FILE_NAME=FA_S10_mitos_no.treatment_KO_2.mzML SUBJECT_SAMPLE_FACTORS - S11_mitos_no.treatment_KO_3 Sample source:total mitochondria | Genotype:FADS2 KO | Condition:untreated RAW_FILE_NAME=-; RAW_FILE_NAME=-; RAW_FILE_NAME=-; RAW_FILE_NAME=FA_S11_mitos_no.treatment_KO_3.mzML SUBJECT_SAMPLE_FACTORS - S12_mitos_no.treatment_KO_4 Sample source:total mitochondria | Genotype:FADS2 KO | Condition:untreated RAW_FILE_NAME=-; RAW_FILE_NAME=-; RAW_FILE_NAME=-; RAW_FILE_NAME=FA_S12_mitos_no.treatment_KO_4.mzML SUBJECT_SAMPLE_FACTORS - S13_mitos_linoleic.acid_KO_1 Sample source:total mitochondria | Genotype:FADS2 KO | Condition:linoleic acid RAW_FILE_NAME=-; RAW_FILE_NAME=-; RAW_FILE_NAME=-; RAW_FILE_NAME=FA_S13_mitos_linoleic.acid_KO_1.mzML SUBJECT_SAMPLE_FACTORS - S14_mitos_linoleic.acid_KO_2 Sample source:total mitochondria | Genotype:FADS2 KO | Condition:linoleic acid RAW_FILE_NAME=-; RAW_FILE_NAME=-; RAW_FILE_NAME=-; RAW_FILE_NAME=FA_S14_mitos_linoleic.acid_KO_2.mzML SUBJECT_SAMPLE_FACTORS - S15_mitos_linoleic.acid_KO_3 Sample source:total mitochondria | Genotype:FADS2 KO | Condition:linoleic acid RAW_FILE_NAME=-; RAW_FILE_NAME=-; RAW_FILE_NAME=-; RAW_FILE_NAME=FA_S15_mitos_linoleic.acid_KO_3.mzML SUBJECT_SAMPLE_FACTORS - S16_mitos_linoleic.acid_KO_4 Sample source:total mitochondria | Genotype:FADS2 KO | Condition:linoleic acid RAW_FILE_NAME=-; RAW_FILE_NAME=-; RAW_FILE_NAME=-; RAW_FILE_NAME=FA_S16_mitos_linoleic.acid_KO_4.mzML #COLLECTION CO:COLLECTION_SUMMARY Human osteosarcoma U2OS WT, U2OS BAK Ko expressing GFP BAK, and U2OS FADS2 KO CO:COLLECTION_SUMMARY cell lines were cultured at 37 °C and 5% CO2 in DMEM supplemented with 10% FBS CO:COLLECTION_SUMMARY and 1% penicillin/streptomycin (Invitrogen, Germany). For lipidomic experiments CO:COLLECTION_SUMMARY cells were incubated with 1 μM of ABT-737 and S63845 in the complete media and CO:COLLECTION_SUMMARY incubated for 50 min at 37°C and 5% CO2. FADS2 KO in U2OS cells was generated CO:COLLECTION_SUMMARY in the lab by the CRISPR/Cas9 method. Linoleic acid stock (50 mM) was prepared CO:COLLECTION_SUMMARY in ethanol and diluted into culture media before adding them to the cells. CO:COLLECTION_SUMMARY Mitochondria were isolated from cultured human osteosarcoma cells by mechanical CO:COLLECTION_SUMMARY disruption of cells followed by differential centrifugation: Cells were CO:COLLECTION_SUMMARY harvested by trypsinization, washed in PBS, and then resuspend in isolation CO:COLLECTION_SUMMARY buffer (IM;250 mM sucrose, 5 mM Tris, and 2 mM EDTA; pH 7.4 and protease CO:COLLECTION_SUMMARY inhibitor cocktail) and mechanically broken using glass homogenizer on ice CO:COLLECTION_SUMMARY (30-40 strokes on ice) and total cellular lysates were spin down first to remove CO:COLLECTION_SUMMARY nuclei and cell debris at 600 x g for 5 min and later at 10,800 x g for 10 min CO:COLLECTION_SUMMARY at 4°C to get the crude mitochondria. Mitochondrial pellet was washed 2-3 times CO:COLLECTION_SUMMARY with isolation buffer to remove other impurities from mitochondria. Isolated CO:COLLECTION_SUMMARY mitochondria were solubilized using SMA co-polymer. For this, mitochondria CO:COLLECTION_SUMMARY either from apoptotic or healthy cells were incubated with 0.5% SMA (2:1) for 45 CO:COLLECTION_SUMMARY min at room temperature with gentle rotation. Mitochondrial membrane was spun CO:COLLECTION_SUMMARY down at 100,000 x g for 40 min to separate solubilized SMALP from the CO:COLLECTION_SUMMARY insolubilized membrane. Next, the size of SMALP was analyzed by Dynamic Light CO:COLLECTION_SUMMARY Scattering (DLS). For DLS measurements, 15 μl of sample was added to a quartz CO:COLLECTION_SUMMARY cuvette which had been thoroughly cleaned with Milli-Q H2O. The cuvette was CO:COLLECTION_SUMMARY placed in DynaPro NanoStar (Wyatt Technology corporation, USA) and the sample CO:COLLECTION_SUMMARY was analyzed using 10 runs with 10 second acquisition time. This helps to CO:COLLECTION_SUMMARY determine the mass distribution of the sample as well as the estimated size of CO:COLLECTION_SUMMARY the particles. The distance distribution is shown on a log scale. The size of CO:COLLECTION_SUMMARY SMALP as well as the homogeneity with in the sample were also checked by CO:COLLECTION_SUMMARY Negative Transmission Electron Microscopy (TEM). For this the diluted SMALPs CO:COLLECTION_SUMMARY were placed onto a glow-discharged copper grid (Electron Microscopy Sciences) CO:COLLECTION_SUMMARY coated with a layer of thin carbon, washed twice with water, stained with 2% CO:COLLECTION_SUMMARY uranyl acetate for 5 min and then air-dried. The grids were imaged on a JEOL CO:COLLECTION_SUMMARY JEM2100PLUS electron microscope and recorded with a GATAN OneView camera (CECAD CO:COLLECTION_SUMMARY Imaging Facility). mEGFP-BAK-SMALPs were affinity purified from total CO:COLLECTION_SUMMARY solubilized mitochondrial membrane fraction (SMALP). For this total SMALP were CO:COLLECTION_SUMMARY incubated with 25 μl of GFP-trap MA beads for 90 min with slow rotation in cold CO:COLLECTION_SUMMARY room. Beads were washed 2 times with 100 μl of Tris buffer (50 mM Tris 150 mM CO:COLLECTION_SUMMARY NaCl pH 8), and finally resuspend in 100 ul of Tris buffer. Small aliquots of CO:COLLECTION_SUMMARY unbound and wash fractions were used to analyze the purification quality. CO:SAMPLE_TYPE Mitochondria #TREATMENT TR:TREATMENT_SUMMARY The samples were not subjected to any further treatment. #SAMPLEPREP SP:SAMPLEPREP_SUMMARY Glycerophospholipids: Lipids from isolated mitochondria treated with or without SP:SAMPLEPREP_SUMMARY SMA were extracted using a procedure previously described (Ejsing et al., 2009) SP:SAMPLEPREP_SUMMARY with some modifications: 30-100 µl of sample were brought to a volume of 200 SP:SAMPLEPREP_SUMMARY µl with 155 mM ammonium carbonate buffer. Lipids were extracted by adding 990 SP:SAMPLEPREP_SUMMARY µl of chloroform/methanol 17:1 (v/v) and internal standards (125 pmol PC SP:SAMPLEPREP_SUMMARY 17:0-20:4, 138 pmol PE 17:0-20:4, 118 pmol PI 17:0-20:4, 118 pmol PS 17:0-20:4, SP:SAMPLEPREP_SUMMARY 61 pmol PG 17:0/20:4, 72 pmol PA 17:0/20:4, 10 µl Cardiolipin Mix I; Avanti SP:SAMPLEPREP_SUMMARY Polar Lipids), followed by shaking at 900 rpm/min in a ThermoMixer (Eppendorf) SP:SAMPLEPREP_SUMMARY at 20 °C for 30 min. After centrifugation (12,000xg, 5 min, 4 °C), the lower SP:SAMPLEPREP_SUMMARY (organic) phase was transferred to a new tube, and the upper phase was extracted SP:SAMPLEPREP_SUMMARY again with 990 mL chloroform/methanol 2:1 (v/v). The combined organic phases SP:SAMPLEPREP_SUMMARY were dried under a stream of nitrogen. The residues were resolved in 200 µl of SP:SAMPLEPREP_SUMMARY methanol. Ceramides and sphingomyelins: For the analysis of ceramides and SP:SAMPLEPREP_SUMMARY sphingomyelins in isolated mitochondria without and after SMA treatment, lipids SP:SAMPLEPREP_SUMMARY were extracted as described above in the presence of 127 pmol ceramide 12:0 and SP:SAMPLEPREP_SUMMARY 124 pmol sphingomyelin 12:0 (internal standards, Avanti Polar Lipids). The dried SP:SAMPLEPREP_SUMMARY extracts were resolved in 100 µL of Milli-Q water and 750 µL of SP:SAMPLEPREP_SUMMARY chloroform/methanol 1:2 (v/v). Alkaline hydrolysis of glycerolipids was SP:SAMPLEPREP_SUMMARY conducted as previously published (Schwamb et al., 2012; Oteng et al., 2017). SP:SAMPLEPREP_SUMMARY Fatty acids: To 100 µl of a suspension of isolated mitochondria in PBS, 500 µl SP:SAMPLEPREP_SUMMARY of methanol, 250 µl of chloroform, and 0.5 µg palmitic-d31 acid SP:SAMPLEPREP_SUMMARY (Sigma-Aldrich) as internal standard were added. The mixture was sonicated for 5 SP:SAMPLEPREP_SUMMARY min, and lipids were extracted in a shaking bath at 48 °C for 1 h. SP:SAMPLEPREP_SUMMARY Glycerolipids were degraded by alkaline hydrolysis adding 75 µl of 1 M SP:SAMPLEPREP_SUMMARY potassium hydroxide in methanol. After 5 min of sonication, the extract was SP:SAMPLEPREP_SUMMARY incubated for 1.5 h at 37 °C, and then neutralized with 6 µl of glacial acetic SP:SAMPLEPREP_SUMMARY acid. 2 ml of chloroform and 4 ml of water were added to the extract which was SP:SAMPLEPREP_SUMMARY vortexed vigorously for 30 sec and then centrifuged (4,000 × g, 5 min, 4 °C) SP:SAMPLEPREP_SUMMARY to separate layers. The lower (organic) phase was transferred to a new tube, and SP:SAMPLEPREP_SUMMARY the upper phase extracted with additional 2 ml of chloroform. The combined SP:SAMPLEPREP_SUMMARY organic phases were dried under a stream of nitrogen. The residues were resolved SP:SAMPLEPREP_SUMMARY in 200 µl of acetonitrile/water 2:1 (v/v) and sonicated for 5 min. After SP:SAMPLEPREP_SUMMARY centrifugation (12,000 × g, 20 min, 4 °C), 40 µl of the clear supernatants SP:SAMPLEPREP_SUMMARY were transferred to autoinjector vials. References: Ejsing et al., Proc Natl SP:SAMPLEPREP_SUMMARY Acad Sci USA 2009, 106, 2136; Oteng et al., J Lipid Res 2017, 58, 1100; Schwamb SP:SAMPLEPREP_SUMMARY et al., Blood 2012, 120, 3978. #CHROMATOGRAPHY CH:CHROMATOGRAPHY_SUMMARY Lipid extract infusion and ionization was conducted using Nano-ESI chips with CH:CHROMATOGRAPHY_SUMMARY the TriVersa NanoMate operated by the ChipSoft Software (Advion) under the CH:CHROMATOGRAPHY_SUMMARY following settings: sample infusion volume: 14 μl, volume of air to aspirate CH:CHROMATOGRAPHY_SUMMARY after sample: 1 μl, air gap before chip: enabled, aspiration delay: 0 s, CH:CHROMATOGRAPHY_SUMMARY prepiercing: with mandrel, spray sensing: enabled, cooling temperature: 14°C, CH:CHROMATOGRAPHY_SUMMARY gas pressure: 0.5 psi, ionization voltage: 1.4 kV, and vent headspace: enabled. CH:CHROMATOGRAPHY_SUMMARY Prewetting was done once. CH:CHROMATOGRAPHY_TYPE None (Direct infusion) CH:INSTRUMENT_NAME Advion TriVersa NanoMate CH:COLUMN_NAME None CH:SOLVENT_A None CH:SOLVENT_B None CH:FLOW_GRADIENT N/A CH:FLOW_RATE N/A CH:COLUMN_TEMPERATURE N/A #ANALYSIS AN:ANALYSIS_TYPE MS #MS MS:INSTRUMENT_NAME SCIEX QTRAP 6500 MS:INSTRUMENT_TYPE QTRAP MS:MS_TYPE ESI MS:ION_MODE POSITIVE MS:MS_COMMENTS PC, PE, PI, PS, PG, and PA species were analyzed by Nano-Electrospray Ionization MS:MS_COMMENTS Tandem Spectrometry (Nano-ESI-MS/MS) with direct infusion of the lipid extract MS:MS_COMMENTS (Shotgun Lipidomics) as previously described (Kumar et al., J Cell Biol 2015, MS:MS_COMMENTS 211, 1057). #MS_METABOLITE_DATA MS_METABOLITE_DATA:UNITS counts per second (cps) MS_METABOLITE_DATA_START Samples S01_mitos_control.SMA_1 S02_mitos_control.SMA_2 S04_mitos_control.SMA_4 S05_mitos_apoptosis.SMA_1 S06_mitos_apoptosis.SMA_2 S07_mitos_apoptosis.SMA_3 S09_mitos_control_1 S11_mitos_control_3 S12_mitos_control_4 S13_mitos_apoptosis_1 S15_mitos_apoptosis_3 S16_mitos_apoptosis_4 S02_pulldown_Control_22.08 S03_pulldown_Control_01.09 S04_pulldown_Control_04.09 S05_pulldown_Control_31.09 S06_pulldown_Apoptosis_22.08 S07_pulldown_Apoptosis_01.09 S08_pulldown_Apoptosis_04.09 S09_pulldown_Apoptosis_31.09 Factors Sample source:mitochondrial SMALPs | Genotype:WT | Condition:control Sample source:mitochondrial SMALPs | Genotype:WT | Condition:control Sample source:mitochondrial SMALPs | Genotype:WT | Condition:control Sample source:mitochondrial SMALPs | Genotype:WT | Condition:apoptotic Sample source:mitochondrial SMALPs | Genotype:WT | Condition:apoptotic Sample source:mitochondrial SMALPs | Genotype:WT | Condition:apoptotic Sample source:total mitochondria | Genotype:WT | Condition:control Sample source:total mitochondria | Genotype:WT | Condition:control Sample source:total mitochondria | Genotype:WT | Condition:control Sample source:total mitochondria | Genotype:WT | Condition:apoptotic Sample source:total mitochondria | Genotype:WT | Condition:apoptotic Sample source:total mitochondria | Genotype:WT | Condition:apoptotic Sample source:mitochondrial SMALPs | Genotype:mEGFP-BAK | Condition:control Sample source:mitochondrial SMALPs | Genotype:mEGFP-BAK | Condition:control Sample source:mitochondrial SMALPs | Genotype:mEGFP-BAK | Condition:control Sample source:mitochondrial SMALPs | Genotype:mEGFP-BAK | Condition:control Sample source:mitochondrial SMALPs | Genotype:mEGFP-BAK | Condition:apoptotic Sample source:mitochondrial SMALPs | Genotype:mEGFP-BAK | Condition:apoptotic Sample source:mitochondrial SMALPs | Genotype:mEGFP-BAK | Condition:apoptotic Sample source:mitochondrial SMALPs | Genotype:mEGFP-BAK | Condition:apoptotic PC 37:4 (IS) 325273882 315726699 34821236 347313432 120047615 138846297 310181654 138121933 287499828 360326534 324784539 299101533 118799333 80605552 125131951 70550146 77719081 97430483 74675266 75210002 PC 30:0 257446577 207274901 24990386 308254725 68377073 112178331 949496558 499247381 604927764 812538364 744888211 815813530 14095862 15028454 25165030 8687986 12906990 14200151 16198909 7492082 PC O-32:0 66326495 62537277 7317646 67721477 23775333 24959251 537950714 215444793 262268359 362688142 306322017 304101677 3041668 9550912 7020587 1981891 3201146 4317851 4834133 2270907 PC O-32:1/P-32:0 187264796 157997984 19549880 239037888 63607376 90011106 449792886 287071678 327465511 443100217 437026729 478392873 9273388 9347555 13433672 4408451 6882225 10619997 8866637 4389316 PC O-32:2/P-32:1 42509658 33849858 4740753 55378503 12960313 19676546 91445733 51413331 60535968 86350915 81927600 98836672 2144681 2410406 3461995 1167360 1780374 2554114 2158959 1043982 PC 32:0 66217033 66790627 8056696 84950256 25510172 21670715 595355102 376135390 389849335 616403470 531212557 504959973 8594219 19930440 16734628 4776073 9297691 8767349 12140308 7051710 PC 32:1 1150996258 783814791 100278594 1410780354 350094638 502686602 2070506376 1218864686 1308082223 1913680622 1982485568 2191376518 79052821 64762865 97804009 42096925 62325041 87972738 65474771 31223190 PC 32:2 245282808 153802873 18799261 305968550 66145333 92248787 374134401 191466385 219198209 336316436 330655456 386739721 14903029 11145845 17068191 7133495 11628901 14218295 11129183 5676742 PC 32:3 18801563 11897629 1557271 24718389 4319045 6742090 28351352 15837835 15780878 23676933 23046404 27339578 1178345 787674 1084374 375831 890442 1000945 844115 398894 PC O-34:0 8073362 14307688 1349566 11758251 5212148 3445710 80741897 52983513 63710725 84131158 70121724 82956720 1117362 2724028 1905939 632556 791066 1129501 1501853 648061 PC O-34:1/P-34:0 290466745 282360692 29283755 383068004 122848976 129755399 664959614 409203113 519547187 691919046 691627687 813150293 12245794 21117964 22446512 6803951 10686527 15041857 14727027 6017098 PC O-34:2/P-34:1 110730623 93241259 10744908 142374582 41888989 48394560 213429227 127632702 145021148 222727724 220576532 268530373 4313853 5272784 7831851 2642983 4629189 6726955 5697081 2699392 PC 34:1 1800089503 1546296296 192003793 2266000412 734187132 931860191 3520260994 2247574478 2501504143 3593113355 3899244463 4308785291 112381011 174711591 165580055 61988261 89086355 138211733 113573541 63232779 PC 34:2 666360024 560118583 57849428 876620516 250204428 319249917 1164155268 684209305 746586598 1079224809 1089334396 1291241180 42428406 70931843 60759328 23289337 32553903 50506271 39550864 21342765 PC 34:3 90169916 65296129 6835579 114667323 27378622 34848819 138188804 76039511 75280390 124694042 121503927 163419692 4962354 3994942 6139957 2235690 3730601 5209748 4117402 2192001 PC 34:4 14079629 13696959 1201329 20853051 5927522 6341534 22539178 14447739 14682960 22240999 27267182 28874150 783396 391647 1173193 401494 748374 867310 922271 315182 PC O-36:1/P-36:0 49478521 55579242 6106083 60528782 20473444 24881649 119085293 86787425 88318861 117972222 125873960 165964481 2526433 7258772 5663781 1648163 2467053 3539989 4678674 1908927 PC O-36:2/P-36:1 111431934 103280406 10289192 142231042 44644669 52628289 193412427 133165227 129936648 197972133 221856417 267731090 4409653 6717421 7721244 2522311 3440228 5871472 4548037 2858760 PC O-36:3/P-36:2 49284861 37050170 5136347 64365626 17371929 21262363 76832580 80688089 51889264 82624655 96850918 166108807 1647168 1935998 2315148 977144 1424385 2211341 1713901 1160563 PC O-36:4/P-36:3 65662488 68416712 5564246 83529153 33255574 30164607 109773805 58803763 70193981 120595434 124738908 156101686 1488459 1735703 2328608 745790 1428332 1893513 1398782 904860 PC 36:0 11382316 8242807 1108420 11915162 3831551 4198164 21015126 13647884 14590528 19914273 18136044 21311466 534525 2581947 514971 203546 402476 492011 538426 238309 PC 36:1 66160226 76986084 6764036 85977955 28690198 41433613 154085220 113438789 114989745 176339985 190216395 239278810 5804511 23040235 9748578 3436058 5355231 7782214 7734680 4025599 PC 36:2 683990263 609596599 72770667 892618421 264496373 338006253 1182309111 763705298 769184668 1160086125 1251188573 1516307706 31946444 57086942 56122282 19137774 28309424 47083390 38091053 21012406 PC 36:3 209582747 191954514 18364809 281749018 66315601 99107811 351264624 197697027 220519599 340517000 416316161 455049437 8908139 16609912 12725268 5041477 7766340 12440402 10209074 5108176 PC 36:4 104397470 122349532 10235724 156338123 55265412 58587864 180566579 124945033 125705332 194516011 243685402 257361126 6821009 7054289 10132049 3248232 6859341 7037141 8117081 3893156 PC 36:5 47634194 51378807 5627719 70985128 22766364 26454704 81353954 52479732 55290341 81602868 97517210 119350126 3913549 2581566 4710365 1617713 3899323 3387886 3654482 2004732 PC 36:6 13693477 14402136 920709 18720974 6224880 5163622 13778112 9528505 9154992 13747451 16845785 20849565 957519 521565 1018468 288806 740052 630081 533898 348481 PC 38:2 28594774 26405335 2910072 39323977 10287563 14287368 38118445 25301507 21286679 30566706 43200029 51479342 1680996 3567272 2472884 633614 954836 2525411 1940662 902906 PC 38:3 61069682 49131040 4786481 71282052 22904645 26620500 86638930 57982568 62601898 80919545 90223766 110539368 2257355 4444288 3803838 1159880 2134336 3475398 2628240 1613618 PC 38:4 64234134 66715421 7538836 89636346 36136527 34013907 102656898 67532348 75813684 118352290 153731257 190528458 3839142 3644195 6675128 2008288 4060917 5351347 4847615 2602581 PC 38:5 96620344 119606702 7944115 142175692 62598389 53781322 168375744 116959008 130132706 176624666 215105342 298516385 5253011 4561283 7437029 2960520 6047843 5599747 6992623 3295293 PC 38:6 55235883 59148510 5301422 74988579 29008247 25247075 91920564 60456579 64168706 87722521 105373730 131074707 3180158 2529906 4697828 1650779 2691949 3505763 3496988 1743162 PC 38:7 11239803 13662487 1069591 19189182 6749133 6235889 19421163 24964567 23592916 32717891 33666770 38815714 487159 1385441 871480 250154 636468 593867 804920 390923 PC 40:3 2964174 2936868 6043925 2029514 2432612 5071358 866629 2843059 4911510 3499950 2885223 191188 950701 310279 48721 103194 319593 225725 159789 PC 40:4 10047147 7086655 11815369 3179689 4555980 13738707 10075604 8404775 13112337 14056430 17405848 261679 584339 628339 247545 423895 519716 322457 229396 PC 40:5 11003644 13836030 1339094 15930480 6164193 6192128 22935738 15627184 14217952 22784730 26943432 34693525 780557 834622 1269463 296504 645386 709055 874539 461821 PC 40:6 33940803 27627611 2758339 50136153 18979304 14367134 57849593 26945795 40008302 49690847 73054090 83428449 1341208 1120001 2274353 721808 1305906 1801391 1885344 786791 PC 40:7 22288865 22517788 2581602 32660792 11910360 12781978 41789430 27403893 28685556 46877267 48433023 59186269 1139702 883148 1678820 549798 978991 1387848 1267808 547447 PC 40:8 5671386 5242498 1014335 7801600 3433306 2851337 12757781 5265153 7616769 12196453 11903793 14995158 390434 699570 416589 344129 458913 373858 319293 PC 40:9 257281 2514957 211879 60983 128244 217736 246570 204770 PE 37:4 (IS) 42203982 48638099 1061263 66798171 4898746 11231877 30144663 11738846 32550662 42316645 25793282 23251602 26055804 19721480 32022146 12802422 21392615 24258873 18892994 11494154 PE 32:0 1017126 1037860 2259878 127913 642100 4220411 1955120 3497145 3955382 2432507 3085523 129847 67450 218686 133042 181227 145699 254407 121352 PE 32:1 10225369 11098980 166226 20588444 1335405 3802951 18570684 7818155 13043956 19998522 12933429 17711735 1454967 1262930 2326863 701368 1353059 2008384 1521872 516910 PE 32:2 2741972 2132249 14544 4751969 234649 569337 3830981 1324120 2098763 3732445 2521705 3074858 147383 130124 440792 101229 261411 386299 405505 85909 PE 32:3 129624 221094 461072 16651 348222 138436 142763 449535 260003 333099 PE O-34:1/P-34:0 2696974 3439289 73439 4728853 449796 926203 5150320 2359547 3751737 5236587 3827490 5389016 382791 250364 955506 200127 361040 421269 718783 PE O-34:2/P-34:1 1451522 1721103 15040 2506122 286559 677407 3528749 1687493 2860485 3876744 3107790 3975038 277060 176329 486728 143478 249354 409257 306120 102512 PE O-34:3/P-34:2 325707 245403 532844 19391 128683 696674 277799 569546 799167 699048 780180 100712 18599 11476 47091 88049 33637 PE 34:1 32336610 36485701 719193 63551758 5158600 12467359 76385771 36905579 50886787 79910147 55298213 72560027 4257661 3831756 7484008 2340369 4487211 6731223 5803974 1952431 PE 34:2 17285145 18706438 325869 33828039 2460226 6428031 30985769 15025913 21303440 31190054 22636608 25614009 2071423 1921554 4565342 1107715 1787814 3444104 3178081 706677 PE 34:3 1881510 1658180 3708564 246146 768132 2876048 1210050 2074640 2920780 2135055 2530912 108867 131597 304598 12096 196515 315097 199033 64249 PE 34:4 540448 831062 1122050 252339 295622 1116047 573556 645040 1396972 990256 934326 17645 42825 110630 10111 110835 64435 94221 9913 PE O-36:1/P-36:0 2471949 3107288 53649 5364399 531765 1008133 6481921 3034125 4892883 5903573 5270470 6946427 375993 285301 588546 194903 403152 577029 633417 139286 PE O-36:2/P-36:1 2545225 3301979 14776 5776474 485567 1056146 5695514 2848692 4066760 5520100 4320472 6436233 445808 346125 1033656 143948 373333 486234 577614 37244 PE O-36:3/P-36:2 944853 1056066 49784 1946601 181106 408859 2166925 865932 1445540 1822040 1710946 2071368 105695 189888 55799 89246 344912 218377 42904 PE O-36:4/P-36:3 746472 787739 1372488 231063 410619 1883518 846383 1157853 1580277 1241368 1537095 50919 31592 219316 23013 155623 154089 135025 12346 PE O-36:5/P-36:4 1392228 2585134 152120 3573331 662454 1401247 4047962 2418622 3625068 4732222 4083688 3903464 255085 160212 344245 142901 479219 649741 710087 125216 PE O-36:6/P-36:5 595762 525463 1118214 43513 234809 1303382 571745 748901 1187109 953183 1191251 118235 19382 205243 22652 126204 63295 77703 PE 36:1 16404980 22043834 539522 34411137 2318236 8287725 45130365 22693669 33133554 46289957 38596412 47551500 2399536 2129362 4153787 1043175 2525964 3500350 3209243 828673 PE 36:2 34606575 41605361 860948 65239730 5229956 15718333 71459387 36245888 52764207 72463899 58798295 78511119 5176907 4927241 9296244 2078063 4321775 7541798 6925828 1750017 PE 36:3 7230949 8852042 12777 14829149 994150 2806451 14240212 6470485 9590438 14436505 10932515 14163920 827494 785353 1420744 350755 905043 1420398 1051138 277157 PE 36:4 8450131 10403858 123801 16427205 1812708 3679106 15063997 7106868 10272805 15532734 12703820 15549950 1026024 671028 1383167 516492 1159466 1366522 1272687 282513 PE 36:5 3662411 3985495 105626 6806020 550304 1184225 5809638 2664597 4008729 6468050 4641764 5761511 479069 263356 525067 91435 419058 617133 434669 192061 PE 36:6 582008 623580 967153 114889 368505 880187 400297 470802 831573 534210 628415 PE O-38:5/P-38:4 1873524 2725072 55643 4454459 410834 843093 4756916 1873566 3826551 4919081 3623885 4486946 162097 214891 455870 117950 313929 413926 500711 59173 PE O-38:6/P-38:5 1858961 2893965 31443 3856185 424482 1109908 5197487 2234037 3715214 5518888 4293171 4965971 340956 162372 430744 115785 300965 309470 316849 133219 PE 38:0 2776554 3735813 169833 4789966 575476 1322220 2900487 1120373 2471091 3978684 2994737 3024328 1071645 1239978 1204382 1033869 931125 1323933 1177502 584288 PE 38:1 567088 1012672 45033 1138592 130847 398348 1789460 850183 1118302 1653906 1485643 2020321 35193 84473 214325 71321 117927 60011 57484 PE 38:2 1846142 2573717 96663 3973956 262542 1032070 5075066 2304176 3861090 5600698 3883572 5280820 248082 163756 436337 88506 149802 450976 460468 117051 PE 38:3 8294370 10960900 139408 15792180 1692184 4090966 19598559 9368576 14812828 20142974 16761785 19572968 1183421 1273826 1952508 421506 1077693 1689837 1124746 299940 PE 38:4 23281528 31439544 544203 51974898 4663948 10115870 50456931 23271944 34843751 50666851 38489134 51052280 3253102 1767065 4834031 1502517 3211713 4100700 3781347 1040275 PE 38:5 16233553 22621150 306256 35436503 3175147 6980084 29134986 14664122 21447698 31852035 24959647 32384696 2246785 1618569 3627328 789849 2126276 2717085 2745709 771964 PE 38:6 4614503 5907778 155484 9325351 634394 1783788 9081570 3284590 5568143 9263535 6589813 9450090 734044 505123 1047427 449868 703609 870132 1108500 194666 PE 38:7 942138 1114307 1973979 284716 487532 1600219 775887 1026545 1442326 1533835 2028765 58866 48671 113874 31506 109648 127844 168931 37102 PE O-40:3/P-40:2 639845 1018020 48691 1328345 197083 369769 1715371 818957 1277091 1662099 1358735 1843644 61517 207889 30497 14865 160680 56475 10253 PE O-40:4/P-40:3 869964 1106643 1634998 67990 332727 1960825 783090 1138350 1734788 1417408 1730886 99476 42118 275006 15239 151330 81175 121670 12699 PE O-40:5/P-40:4 477570 1251422 1629364 222821 351382 1563620 842844 1169726 1791650 1425273 1776651 81254 93321 189932 28568 149837 91049 53568 PE O-40:6/P-40:5 658993 999140 1402778 121574 223209 1898418 826060 1068622 2268190 1389771 2187257 20313 44518 155696 91114 25558 55890 PE 40:3 887051 729773 15700 1504290 20483 392361 1647080 1033374 1526740 2106881 1828744 2238786 PE 40:4 1584197 1852382 2733324 182155 941640 2752669 1562746 2441590 3221032 2395877 3358847 90959 86635 258131 25118 78935 167537 111137 46877 PE 40:5 2959176 3828502 120674 5760086 397724 972337 6074327 2792142 4205033 6447119 4926222 6330405 355998 216110 488186 125475 506817 489150 441902 84343 PE 40:6 4217796 5523004 78454 8899808 877053 1893352 8666448 4366224 6308767 9956672 7114491 10855921 682854 377768 1010890 267389 516678 875481 651567 131416 PE 40:7 3368875 4326509 44305 6625303 787756 1839972 5636280 3288915 4058014 6201816 5418797 7353466 402254 347236 828601 267952 294937 771857 458953 165986 PE 40:8 1104321 1727613 59346 2289170 441960 626348 1577189 902342 1547140 1990241 2115917 2505888 137344 43450 292506 77574 107035 136426 255890 12929 PE 40:9 547172 608406 807269 189651 241380 932341 261174 584876 807561 758377 860150 19386 134782 16104 34752 13053 63994 PI 37:4 (IS) 23990720 30442855 195317 51837436 2840409 5173459 19007712 4927893 20048987 29936090 13939449 10735364 21725823 17323827 29760541 13397328 18230602 22704112 15428604 14880811 PI 32:0 802965 1147267 413830 2103400 1338043 1726303 591803 1478782 1148676 1493162 156877 PI 32:1 2165503 1342230 297788 2960828 1246375 2070816 1992800 744062 1370902 2026108 2347333 1744230 165937 71379 134782 147258 151553 112842 56040 PI 32:2 886594 718777 193906 1183350 1055735 1659416 586591 91902 900136 861915 1009350 889930 PI 32:3 297339 253905 173772 457021 245765 662582 335891 41820 268631 48080 638254 454165 PI 34:1 3754959 3559607 9991551 480484 1165288 6994426 3314116 4779721 8457241 5229135 5750101 377916 450913 624500 237437 485831 745584 507424 188601 PI 34:2 4444155 3701149 26353 9458370 375459 823159 5874470 2521094 3573595 6582642 4225058 4605796 481045 485437 812966 259137 445280 594792 544627 203921 PI 34:3 759652 610885 1421654 273911 382823 808220 255117 492935 709074 416683 428915 55415 40360 31158 90755 59386 45796 PI 34:4 484572 369083 15117 691645 256020 273492 314183 34347 294301 294530 193844 366079 PI 36:1 3165550 3905079 8103660 576598 6394009 3072142 5199318 7813868 5440055 6244478 547073 509602 762141 362346 399970 656115 711519 174338 PI 36:2 7526359 8286758 87725 18669107 1256825 1861535 13039933 4675864 9484861 14227690 10862850 11139037 862855 1048940 1726096 566896 937777 1988810 1700466 567027 PI 36:3 3701172 2682981 8880413 362135 792296 5357370 2228205 3876746 6195971 3761510 4529026 360559 468397 565621 197478 414841 629792 551490 174916 PI 36:4 2210288 3738157 66503 6923538 780786 1429865 4567489 1836484 3302188 4239217 3457710 3262368 329096 271726 685130 221322 443472 644340 460895 239070 PI 36:5 410345 299212 903936 162051 302357 821173 364508 530629 947746 730356 580283 PI 38:1 538261 155795 366870 226912 203326 253713 309674 252059 330770 315585 PI 38:2 3131353 2176015 40565 6724917 541086 998957 4369564 2010578 3772525 5775713 2902094 3827442 327210 442012 661143 316703 205415 849064 413171 127634 PI 38:3 15350011 13622768 167092 33729360 1449635 3123771 26571122 9592116 16413644 27214224 16819948 19902390 1671210 1955433 3342593 1093117 1847848 3082835 1932746 988153 PI 38:4 13575166 18814885 148060 33033792 1954961 3542685 27606777 10344657 19567171 28064742 19631488 24041332 3000424 2049266 5630858 1499225 3096080 4147796 3692086 1427633 PI 38:5 4325064 5766274 16400 11749541 659049 1110239 8024924 3086379 5932202 9475739 5985657 7633167 1155764 940611 2244254 682217 1399841 1683140 1452575 582080 PI 38:6 750210 647911 1620646 51811 200249 1073601 458649 543341 1136309 586964 893688 223422 66895 234571 118051 130109 60839 60993 PI 40:3 737330 687108 1631136 92173 224611 1508554 624817 864321 1540488 1093432 1213416 PI 40:4 1126100 809317 2407282 79633 413689 1942819 702602 1349476 2003213 1319651 1252027 265361 174655 320958 246736 198273 56682 PI 40:5 1860485 1742274 4089888 321610 563261 2453334 1040031 1531148 2951813 1839000 1888608 384080 318508 541892 114077 292768 526579 552456 128268 PI 40:6 1822467 1777701 4146264 141021 574475 2516987 988632 1773351 2665855 1928916 2551752 240246 306837 498412 198099 290689 521771 364045 152487 PS 37:4 (IS) 33488402 40569952 10097 65235863 1782894 5768841 21929284 5629739 22581430 27797965 12746600 9969232 23911793 17778407 31274238 13780037 19927179 21403064 16113999 14287263 PS 32:0 303074 112688 79275 39609 66118 64800 287112 PS 32:2 468215 607723 1125995 249785 304582 817334 327708 559835 1067920 533700 560284 204145 184953 128183 426290 242944 400021 216105 125209 PS 32:3 291198 499805 374952 206164 339778 385435 201142 302904 597882 602155 495680 162802 163738 33488 89809 174479 149044 63030 44125 PS 34:1 616063 1041273 558135 1301039 250983 417758 277459 PS 34:2 2124084 2511934 13300 4457789 286659 694254 7480769 3269632 5392823 6997654 3907718 4373374 PS 34:3 336463 468478 517141 39130 342306 710132 398356 654976 966000 395326 671516 PS 34:4 621681 672524 749491 204314 299718 1128160 389134 617190 910012 783065 1018871 34259 17256 81784 81361 66162 52234 PS 36:1 12418915 19178631 38099 29223360 1102404 3861882 80166305 31343985 57803440 69373199 45263850 50604979 2941538 1571911 6103815 1782004 4024292 3136836 3607696 1693395 PS 36:2 652991 1331756 272810 1016027 786275 767437 450269 PS 36:3 980231 1370131 16798 2388705 130261 476567 3248119 1289452 2619257 3138775 2109004 2322552 PS 36:4 1124777 1419489 1541386 416719 685957 2599004 1152653 2290653 2586062 2450725 2181186 PS 36:5 283804 641818 671073 109004 145771 876630 267998 525935 915488 818700 384366 PS 36:6 220884 144851 62811 112646 268483 41851 40336 383808 92644 54188 PS 38:1 614338 848941 40692 892264 155935 206759 1676659 564766 868156 1561884 1545027 1286638 95597 39023 200198 43556 85020 95206 110320 PS 38:2 919888 1099803 2390683 101492 1660542 215538 1376289 1338285 PS 38:3 1647859 3268862 5158573 440731 4907253 1216409 4962188 4807914 1820931 2288888 543571 303274 828178 198314 585027 433449 481243 236730 PS 38:4 332693 272119 770087 348834 383637 465730 479828 227693 PS 38:5 39300 87467 250296 117422 145642 60334 54836 133796 PS 38:7 191806 310448 383397 14145 36860 361966 186983 363971 504719 179747 342724 PS 40:1 475222 869145 1176609 38029 234513 2167419 800967 1169791 1735184 1041696 1001684 86281 73644 204353 70638 116235 51456 104925 92997 PS 40:2 702916 805595 1591437 60622 230306 1920075 731783 1109959 1761626 1087291 1282815 119442 116955 141861 98472 79719 163706 119846 37435 PS 40:3 392726 894322 1203513 137993 1592891 445856 1458546 1138862 453347 992341 136349 124609 12123 60975 113748 57124 PS 40:4 654129 882884 1222916 196278 2171399 759727 1277060 1830493 500673 683838 114518 101173 264191 99988 226134 130829 100673 PS 40:5 2094050 2871258 9924 4306738 129826 489982 6883387 3324136 5061656 5871959 4348570 4610643 445054 135567 454604 91451 388827 260963 302037 222949 PS 40:6 1707764 2177773 3431525 673153 6705111 2949941 5101169 5409880 3562516 4276381 483032 165253 590259 226890 365501 400005 467324 162741 PS 40:7 452829 593089 914465 93538 298972 1031603 632138 581453 1156086 724719 876284 PS 40:8 948508 1381703 118658 945755 592643 914645 1214969 330740 1014242 1867045 1033487 1080404 PS 40:9 218902 244837 320621 51050 122730 216220 48170 22441 12200 PG 37:4 (IS) 16653049 19579171 200126 32563887 2051918 3737411 13390331 3066044 13039564 20157960 8476736 7447858 31510151 21066418 41536400 19487412 24960994 28647347 20345215 19373024 PG 32:0 390941 908605 42422 1173982 227804 391664 23140970 13396427 23243415 31674976 15874418 15000864 949629 375509 1192170 535801 671476 900368 702881 421082 PG 32:1 443635 521342 869463 41062 172735 1268430 564513 1066603 1639780 884920 906744 297078 235227 477916 323624 6681 225199 281665 97979 PG 32:2 192502 238892 335949 67676 312726 56342 259286 304865 70440 71442 76782 130352 50215 120732 43044 167827 41752 39655 PG 32:3 69360 127582 285086 54211 88548 187664 207365 182204 151455 52602 20252 35722 91627 148973 136345 64574 47695 PG 34:0 64986 126280 35466 177268 42189 115424 PG 34:1 3517308 4351735 60448 7799911 433909 1151677 23881508 12259128 24512939 33428195 18744151 18016988 1470842 961465 2000990 780134 1194536 1835139 1200106 529977 PG 34:2 553287 713808 1173789 75581 121785 1235817 561000 892423 1544658 969214 981986 185126 102024 510000 86339 163041 191259 388639 42768 PG 34:3 83976 131291 194414 13651 32104 79539 32955 82550 139749 113589 116070 PG 36:0 436267 549029 406355 178790 239008 874317 409167 918999 1648473 1346271 1392048 329440 269529 160899 160504 135135 130167 121678 212217 PG 36:1 650027 912111 11913 2297680 315774 11499871 5985989 10547232 16600490 8130945 8437554 202303 183706 277154 94739 306605 399199 350756 31987 PG 36:2 378504 710420 1284408 87064 173428 4120262 1919599 3641670 5593113 2946130 2984034 921462 620811 1362275 501701 739203 1027039 725751 558471 PG 36:3 130457 142940 509239 41107 513278 161606 391919 706150 208688 400461 PG 36:4 168326 630258 1209160 76773 249409 400081 133223 366273 693992 660782 190080 135482 84368 128502 36744 47008 134455 58627 35513 PG 38:2 81876 192591 643822 15123 321297 49263 351636 755621 233560 379581 713707 412934 938790 498232 649886 658258 473013 548408 PG 38:3 3361 308824 375061 35698 607674 111991 315393 756220 295085 419471 PG 38:4 101196 20575 148015 33666 72013 52907 45139 25194 PG 38:5 31697 227851 289261 24350 245828 62337 207184 325699 131432 238044 PG 40:3 10377 89200 114270 35655 150812 8342 26828 268455 146255 185405 PG 40:9 63285 119214 290424 42031 221604 106845 121601 33766 229017 172809 161731 PA 37:4 (IS) 20590829 14575844 25470722 12432876 16287621 19973694 13428083 10776113 PA 30:1 124226 75281 19025 89123 65798 22489 117592 114337 PA 30:2 126075 44325 109335 82371 131743 176798 PA 32:0 81992 240620 180034 168672 164172 336805 234625 212486 PA 32:1 172124 77858 303371 291853 199151 164437 425004 367612 PA 32:2 154871 130296 117686 196429 38722 298064 326044 125887 PA 32:3 98785 5844 52878 59488 18376 148836 45803 22473 PA 34:1 371719 355254 855220 441615 326176 471571 820690 602231 PA 34:2 141565 179385 339044 326577 113104 330154 495733 379720 PA 34:3 108937 17096 45004 104570 61615 40562 187881 43834 PA 36:0 6305533 3150223 4199590 2413657 2298458 3375087 2427191 4412646 PA 36:1 255632 121690 299424 162127 229390 173739 351322 85020 PA 36:2 278949 326788 744456 283339 253660 427064 408975 387921 PA 36:3 118987 124822 84215 95181 100427 192124 68747 PA 36:6 8987 19545 12635 10661 39485 35734 PA 38:2 149141 120669 173687 103705 183495 97856 129321 PA 38:3 98600 66070 120708 18249 110689 113062 163069 38428 PA 38:4 81815 144843 114945 111713 148691 70784 144040 91512 PA 40:8 11731 95581 28901 15276 48219 8006 PA 40:9 76574 104232 75749 17750 157483 51161 MS_METABOLITE_DATA_END #METABOLITES METABOLITES_START metabolite_name Mass to Charge (m/z) Fragmentation Modification Charge KEGG_ID LM_ID PC 37:4 (IS) 796.5 +Precursor Ion Scan m/z 184 [M+H] 1 C00157 LMGP01010000 PC 30:0 706.5 +Precursor Ion Scan m/z 184 [M+H] 1 C00157 LMGP01010000 PC O-32:0 720.5 +Precursor Ion Scan m/z 184 [M+H] 1 C00157 LMGP01010000 PC O-32:1/P-32:0 718.5 +Precursor Ion Scan m/z 184 [M+H] 1 C05212/C00958 LMGP01020000/LMGP01030000 PC O-32:2/P-32:1 716.5 +Precursor Ion Scan m/z 184 [M+H] 1 C05212/C00958 LMGP01020000/LMGP01030000 PC 32:0 734.5 +Precursor Ion Scan m/z 184 [M+H] 1 C00157 LMGP01010000 PC 32:1 732.5 +Precursor Ion Scan m/z 184 [M+H] 1 C00157 LMGP01010000 PC 32:2 730.5 +Precursor Ion Scan m/z 184 [M+H] 1 C00157 LMGP01010000 PC 32:3 728.5 +Precursor Ion Scan m/z 184 [M+H] 1 C00157 LMGP01010000 PC O-34:0 748.5 +Precursor Ion Scan m/z 184 [M+H] 1 C00157 LMGP01010000 PC O-34:1/P-34:0 746.5 +Precursor Ion Scan m/z 184 [M+H] 1 C05212/C00958 LMGP01020000/LMGP01030000 PC O-34:2/P-34:1 744.5 +Precursor Ion Scan m/z 184 [M+H] 1 C05212/C00958 LMGP01020000/LMGP01030000 PC 34:1 760.5 +Precursor Ion Scan m/z 184 [M+H] 1 C00157 LMGP01010000 PC 34:2 758.5 +Precursor Ion Scan m/z 184 [M+H] 1 C00157 LMGP01010000 PC 34:3 756.5 +Precursor Ion Scan m/z 184 [M+H] 1 C00157 LMGP01010000 PC 34:4 754.5 +Precursor Ion Scan m/z 184 [M+H] 1 C00157 LMGP01010000 PC O-36:1/P-36:0 774.5 +Precursor Ion Scan m/z 184 [M+H] 1 C05212/C00958 LMGP01020000/LMGP01030000 PC O-36:2/P-36:1 772.5 +Precursor Ion Scan m/z 184 [M+H] 1 C05212/C00958 LMGP01020000/LMGP01030000 PC O-36:3/P-36:2 770.5 +Precursor Ion Scan m/z 184 [M+H] 1 C05212/C00958 LMGP01020000/LMGP01030000 PC O-36:4/P-36:3 768.5 +Precursor Ion Scan m/z 184 [M+H] 1 C05212/C00958 LMGP01020000/LMGP01030000 PC 36:0 790.5 +Precursor Ion Scan m/z 184 [M+H] 1 C00157 LMGP01010000 PC 36:1 788.5 +Precursor Ion Scan m/z 184 [M+H] 1 C00157 LMGP01010000 PC 36:2 786.5 +Precursor Ion Scan m/z 184 [M+H] 1 C00157 LMGP01010000 PC 36:3 784.5 +Precursor Ion Scan m/z 184 [M+H] 1 C00157 LMGP01010000 PC 36:4 782.5 +Precursor Ion Scan m/z 184 [M+H] 1 C00157 LMGP01010000 PC 36:5 780.5 +Precursor Ion Scan m/z 184 [M+H] 1 C00157 LMGP01010000 PC 36:6 778.5 +Precursor Ion Scan m/z 184 [M+H] 1 C00157 LMGP01010000 PC 38:2 814.5 +Precursor Ion Scan m/z 184 [M+H] 1 C00157 LMGP01010000 PC 38:3 812.5 +Precursor Ion Scan m/z 184 [M+H] 1 C00157 LMGP01010000 PC 38:4 810.5 +Precursor Ion Scan m/z 184 [M+H] 1 C00157 LMGP01010000 PC 38:5 808.5 +Precursor Ion Scan m/z 184 [M+H] 1 C00157 LMGP01010000 PC 38:6 806.5 +Precursor Ion Scan m/z 184 [M+H] 1 C00157 LMGP01010000 PC 38:7 804.5 +Precursor Ion Scan m/z 184 [M+H] 1 C00157 LMGP01010000 PC 40:3 840.5 +Precursor Ion Scan m/z 184 [M+H] 1 C00157 LMGP01010000 PC 40:4 838.5 +Precursor Ion Scan m/z 184 [M+H] 1 C00157 LMGP01010000 PC 40:5 836.5 +Precursor Ion Scan m/z 184 [M+H] 1 C00157 LMGP01010000 PC 40:6 834.5 +Precursor Ion Scan m/z 184 [M+H] 1 C00157 LMGP01010000 PC 40:7 832.5 +Precursor Ion Scan m/z 184 [M+H] 1 C00157 LMGP01010000 PC 40:8 830.5 +Precursor Ion Scan m/z 184 [M+H] 1 C00157 LMGP01010000 PC 40:9 828.5 +Precursor Ion Scan m/z 184 [M+H] 1 C00157 LMGP01010000 PE 37:4 (IS) 754.4 +Neutral Loss Scan m/z 141 [M+H] 1 C00350 LMGP02010000 PE 32:0 692.4 +Neutral Loss Scan m/z 141 [M+H] 1 C00350 LMGP02010000 PE 32:1 690.4 +Neutral Loss Scan m/z 141 [M+H] 1 C00350 LMGP02010000 PE 32:2 688.4 +Neutral Loss Scan m/z 141 [M+H] 1 C00350 LMGP02010000 PE 32:3 686.4 +Neutral Loss Scan m/z 141 [M+H] 1 C00350 LMGP02010000 PE O-34:1/P-34:0 704.4 +Neutral Loss Scan m/z 141 [M+H] 1 C04475/C04756 LMGP02020000/LMGP02030000 PE O-34:2/P-34:1 702.4 +Neutral Loss Scan m/z 141 [M+H] 1 C04475/C04756 LMGP02020000/LMGP02030000 PE O-34:3/P-34:2 700.4 +Neutral Loss Scan m/z 141 [M+H] 1 C04475/C04756 LMGP02020000/LMGP02030000 PE 34:1 718.5 +Neutral Loss Scan m/z 141 [M+H] 1 C00350 LMGP02010000 PE 34:2 716.5 +Neutral Loss Scan m/z 141 [M+H] 1 C00350 LMGP02010000 PE 34:3 714.5 +Neutral Loss Scan m/z 141 [M+H] 1 C00350 LMGP02010000 PE 34:4 712.5 +Neutral Loss Scan m/z 141 [M+H] 1 C00350 LMGP02010000 PE O-36:1/P-36:0 732.5 +Neutral Loss Scan m/z 141 [M+H] 1 C04475/C04756 LMGP02020000/LMGP02030000 PE O-36:2/P-36:1 730.5 +Neutral Loss Scan m/z 141 [M+H] 1 C04475/C04756 LMGP02020000/LMGP02030000 PE O-36:3/P-36:2 728.5 +Neutral Loss Scan m/z 141 [M+H] 1 C04475/C04756 LMGP02020000/LMGP02030000 PE O-36:4/P-36:3 726.5 +Neutral Loss Scan m/z 141 [M+H] 1 C04475/C04756 LMGP02020000/LMGP02030000 PE O-36:5/P-36:4 724.5 +Neutral Loss Scan m/z 141 [M+H] 1 C04475/C04756 LMGP02020000/LMGP02030000 PE O-36:6/P-36:5 722.5 +Neutral Loss Scan m/z 141 [M+H] 1 C04475/C04756 LMGP02020000/LMGP02030000 PE 36:1 746.5 +Neutral Loss Scan m/z 141 [M+H] 1 C00350 LMGP02010000 PE 36:2 744.5 +Neutral Loss Scan m/z 141 [M+H] 1 C00350 LMGP02010000 PE 36:3 742.5 +Neutral Loss Scan m/z 141 [M+H] 1 C00350 LMGP02010000 PE 36:4 740.5 +Neutral Loss Scan m/z 141 [M+H] 1 C00350 LMGP02010000 PE 36:5 738.5 +Neutral Loss Scan m/z 141 [M+H] 1 C00350 LMGP02010000 PE 36:6 736.5 +Neutral Loss Scan m/z 141 [M+H] 1 C00350 LMGP02010000 PE O-38:5/P-38:4 752.5 +Neutral Loss Scan m/z 141 [M+H] 1 C04475/C04756 LMGP02020000/LMGP02030000 PE O-38:6/P-38:5 750.5 +Neutral Loss Scan m/z 141 [M+H] 1 C04475/C04756 LMGP02020000/LMGP02030000 PE 38:0 776.5 +Neutral Loss Scan m/z 141 [M+H] 1 C00350 LMGP02010000 PE 38:1 774.5 +Neutral Loss Scan m/z 141 [M+H] 1 C00350 LMGP02010000 PE 38:2 772.5 +Neutral Loss Scan m/z 141 [M+H] 1 C00350 LMGP02010000 PE 38:3 770.5 +Neutral Loss Scan m/z 141 [M+H] 1 C00350 LMGP02010000 PE 38:4 768.5 +Neutral Loss Scan m/z 141 [M+H] 1 C00350 LMGP02010000 PE 38:5 766.5 +Neutral Loss Scan m/z 141 [M+H] 1 C00350 LMGP02010000 PE 38:6 764.5 +Neutral Loss Scan m/z 141 [M+H] 1 C00350 LMGP02010000 PE 38:7 762.5 +Neutral Loss Scan m/z 141 [M+H] 1 C00350 LMGP02010000 PE O-40:3/P-40:2 784.5 +Neutral Loss Scan m/z 141 [M+H] 1 C04475/C04756 LMGP02020000/LMGP02030000 PE O-40:4/P-40:3 782.5 +Neutral Loss Scan m/z 141 [M+H] 1 C04475/C04756 LMGP02020000/LMGP02030000 PE O-40:5/P-40:4 780.5 +Neutral Loss Scan m/z 141 [M+H] 1 C04475/C04756 LMGP02020000/LMGP02030000 PE O-40:6/P-40:5 778.5 +Neutral Loss Scan m/z 141 [M+H] 1 C04475/C04756 LMGP02020000/LMGP02030000 PE 40:3 798.5 +Neutral Loss Scan m/z 141 [M+H] 1 C00350 LMGP02010000 PE 40:4 796.5 +Neutral Loss Scan m/z 141 [M+H] 1 C00350 LMGP02010000 PE 40:5 794.5 +Neutral Loss Scan m/z 141 [M+H] 1 C00350 LMGP02010000 PE 40:6 792.5 +Neutral Loss Scan m/z 141 [M+H] 1 C00350 LMGP02010000 PE 40:7 790.5 +Neutral Loss Scan m/z 141 [M+H] 1 C00350 LMGP02010000 PE 40:8 788.5 +Neutral Loss Scan m/z 141 [M+H] 1 C00350 LMGP02010000 PE 40:9 786.5 +Neutral Loss Scan m/z 141 [M+H] 1 C00350 LMGP02010000 PI 37:4 (IS) 890.5 +Neutral Loss Scan m/z 277 [M+NH4] 1 C01194 LMGP06010000 PI 32:0 828.5 +Neutral Loss Scan m/z 277 [M+NH4] 1 C01194 LMGP06010000 PI 32:1 826.5 +Neutral Loss Scan m/z 277 [M+NH4] 1 C01194 LMGP06010000 PI 32:2 824.5 +Neutral Loss Scan m/z 277 [M+NH4] 1 C01194 LMGP06010000 PI 32:3 822.5 +Neutral Loss Scan m/z 277 [M+NH4] 1 C01194 LMGP06010000 PI 34:1 854.5 +Neutral Loss Scan m/z 277 [M+NH4] 1 C01194 LMGP06010000 PI 34:2 852.5 +Neutral Loss Scan m/z 277 [M+NH4] 1 C01194 LMGP06010000 PI 34:3 850.5 +Neutral Loss Scan m/z 277 [M+NH4] 1 C01194 LMGP06010000 PI 34:4 848.5 +Neutral Loss Scan m/z 277 [M+NH4] 1 C01194 LMGP06010000 PI 36:1 882.5 +Neutral Loss Scan m/z 277 [M+NH4] 1 C01194 LMGP06010000 PI 36:2 880.5 +Neutral Loss Scan m/z 277 [M+NH4] 1 C01194 LMGP06010000 PI 36:3 878.5 +Neutral Loss Scan m/z 277 [M+NH4] 1 C01194 LMGP06010000 PI 36:4 876.5 +Neutral Loss Scan m/z 277 [M+NH4] 1 C01194 LMGP06010000 PI 36:5 874.5 +Neutral Loss Scan m/z 277 [M+NH4] 1 C01194 LMGP06010000 PI 38:1 910.5 +Neutral Loss Scan m/z 277 [M+NH4] 1 C01194 LMGP06010000 PI 38:2 908.5 +Neutral Loss Scan m/z 277 [M+NH4] 1 C01194 LMGP06010000 PI 38:3 906.5 +Neutral Loss Scan m/z 277 [M+NH4] 1 C01194 LMGP06010000 PI 38:4 904.5 +Neutral Loss Scan m/z 277 [M+NH4] 1 C01194 LMGP06010000 PI 38:5 902.5 +Neutral Loss Scan m/z 277 [M+NH4] 1 C01194 LMGP06010000 PI 38:6 900.5 +Neutral Loss Scan m/z 277 [M+NH4] 1 C01194 LMGP06010000 PI 40:3 934.5 +Neutral Loss Scan m/z 277 [M+NH4] 1 C01194 LMGP06010000 PI 40:4 932.5 +Neutral Loss Scan m/z 277 [M+NH4] 1 C01194 LMGP06010000 PI 40:5 930.5 +Neutral Loss Scan m/z 277 [M+NH4] 1 C01194 LMGP06010000 PI 40:6 928.5 +Neutral Loss Scan m/z 277 [M+NH4] 1 C01194 LMGP06010000 PS 37:4 (IS) 798.5 +Neutral Loss Scan m/z 185 [M+H] 1 C02737 LMGP03010000 PS 32:0 736.5 +Neutral Loss Scan m/z 185 [M+H] 1 C02737 LMGP03010000 PS 32:2 732.5 +Neutral Loss Scan m/z 185 [M+H] 1 C02737 LMGP03010000 PS 32:3 730.5 +Neutral Loss Scan m/z 185 [M+H] 1 C02737 LMGP03010000 PS 34:1 762.5 +Neutral Loss Scan m/z 185 [M+H] 1 C02737 LMGP03010000 PS 34:2 760.5 +Neutral Loss Scan m/z 185 [M+H] 1 C02737 LMGP03010000 PS 34:3 758.5 +Neutral Loss Scan m/z 185 [M+H] 1 C02737 LMGP03010000 PS 34:4 756.5 +Neutral Loss Scan m/z 185 [M+H] 1 C02737 LMGP03010000 PS 36:1 790.5 +Neutral Loss Scan m/z 185 [M+H] 1 C02737 LMGP03010000 PS 36:2 788.5 +Neutral Loss Scan m/z 185 [M+H] 1 C02737 LMGP03010000 PS 36:3 786.5 +Neutral Loss Scan m/z 185 [M+H] 1 C02737 LMGP03010000 PS 36:4 784.5 +Neutral Loss Scan m/z 185 [M+H] 1 C02737 LMGP03010000 PS 36:5 782.5 +Neutral Loss Scan m/z 185 [M+H] 1 C02737 LMGP03010000 PS 36:6 780.5 +Neutral Loss Scan m/z 185 [M+H] 1 C02737 LMGP03010000 PS 38:1 818.5 +Neutral Loss Scan m/z 185 [M+H] 1 C02737 LMGP03010000 PS 38:2 816.5 +Neutral Loss Scan m/z 185 [M+H] 1 C02737 LMGP03010000 PS 38:3 814.5 +Neutral Loss Scan m/z 185 [M+H] 1 C02737 LMGP03010000 PS 38:4 812.5 +Neutral Loss Scan m/z 185 [M+H] 1 C02737 LMGP03010000 PS 38:5 810.5 +Neutral Loss Scan m/z 185 [M+H] 1 C02737 LMGP03010000 PS 38:7 806.5 +Neutral Loss Scan m/z 185 [M+H] 1 C02737 LMGP03010000 PS 40:1 846.5 +Neutral Loss Scan m/z 185 [M+H] 1 C02737 LMGP03010000 PS 40:2 844.5 +Neutral Loss Scan m/z 185 [M+H] 1 C02737 LMGP03010000 PS 40:3 842.5 +Neutral Loss Scan m/z 185 [M+H] 1 C02737 LMGP03010000 PS 40:4 840.5 +Neutral Loss Scan m/z 185 [M+H] 1 C02737 LMGP03010000 PS 40:5 838.5 +Neutral Loss Scan m/z 185 [M+H] 1 C02737 LMGP03010000 PS 40:6 836.5 +Neutral Loss Scan m/z 185 [M+H] 1 C02737 LMGP03010000 PS 40:7 834.5 +Neutral Loss Scan m/z 185 [M+H] 1 C02737 LMGP03010000 PS 40:8 832.5 +Neutral Loss Scan m/z 185 [M+H] 1 C02737 LMGP03010000 PS 40:9 830.5 +Neutral Loss Scan m/z 185 [M+H] 1 C02737 LMGP03010000 PG 37:4 (IS) 802.5 +Neutral Loss Scan m/z 189 [M+NH4] 1 C00344 LMGP04010000 PG 32:0 740.4 +Neutral Loss Scan m/z 189 [M+NH4] 1 C00344 LMGP04010000 PG 32:1 738.4 +Neutral Loss Scan m/z 189 [M+NH4] 1 C00344 LMGP04010000 PG 32:2 736.4 +Neutral Loss Scan m/z 189 [M+NH4] 1 C00344 LMGP04010000 PG 32:3 734.4 +Neutral Loss Scan m/z 189 [M+NH4] 1 C00344 LMGP04010000 PG 34:0 768.4 +Neutral Loss Scan m/z 189 [M+NH4] 1 C00344 LMGP04010000 PG 34:1 766.4 +Neutral Loss Scan m/z 189 [M+NH4] 1 C00344 LMGP04010000 PG 34:2 764.4 +Neutral Loss Scan m/z 189 [M+NH4] 1 C00344 LMGP04010000 PG 34:3 762.4 +Neutral Loss Scan m/z 189 [M+NH4] 1 C00344 LMGP04010000 PG 36:0 796.5 +Neutral Loss Scan m/z 189 [M+NH4] 1 C00344 LMGP04010000 PG 36:1 794.5 +Neutral Loss Scan m/z 189 [M+NH4] 1 C00344 LMGP04010000 PG 36:2 792.4 +Neutral Loss Scan m/z 189 [M+NH4] 1 C00344 LMGP04010000 PG 36:3 790.4 +Neutral Loss Scan m/z 189 [M+NH4] 1 C00344 LMGP04010000 PG 36:4 788.4 +Neutral Loss Scan m/z 189 [M+NH4] 1 C00344 LMGP04010000 PG 38:2 820.5 +Neutral Loss Scan m/z 189 [M+NH4] 1 C00344 LMGP04010000 PG 38:3 818.5 +Neutral Loss Scan m/z 189 [M+NH4] 1 C00344 LMGP04010000 PG 38:4 816.5 +Neutral Loss Scan m/z 189 [M+NH4] 1 C00344 LMGP04010000 PG 38:5 814.5 +Neutral Loss Scan m/z 189 [M+NH4] 1 C00344 LMGP04010000 PG 40:3 846.5 +Neutral Loss Scan m/z 189 [M+NH4] 1 C00344 LMGP04010000 PG 40:9 834.5 +Neutral Loss Scan m/z 189 [M+NH4] 1 C00344 LMGP04010000 PA 37:4 (IS) 728.5 +Neutral Loss Scan m/z 115 [M+NH4] 1 C00416 LMGP10010000 PA 30:1 636.4 +Neutral Loss Scan m/z 115 [M+NH4] 1 C00416 LMGP10010000 PA 30:2 634.4 +Neutral Loss Scan m/z 115 [M+NH4] 1 C00416 LMGP10010000 PA 32:0 666.4 +Neutral Loss Scan m/z 115 [M+NH4] 1 C00416 LMGP10010000 PA 32:1 664.4 +Neutral Loss Scan m/z 115 [M+NH4] 1 C00416 LMGP10010000 PA 32:2 662.4 +Neutral Loss Scan m/z 115 [M+NH4] 1 C00416 LMGP10010000 PA 32:3 660.4 +Neutral Loss Scan m/z 115 [M+NH4] 1 C00416 LMGP10010000 PA 34:1 692.4 +Neutral Loss Scan m/z 115 [M+NH4] 1 C00416 LMGP10010000 PA 34:2 690.4 +Neutral Loss Scan m/z 115 [M+NH4] 1 C00416 LMGP10010000 PA 34:3 688.4 +Neutral Loss Scan m/z 115 [M+NH4] 1 C00416 LMGP10010000 PA 36:0 722.5 +Neutral Loss Scan m/z 115 [M+NH4] 1 C00416 LMGP10010000 PA 36:1 720.5 +Neutral Loss Scan m/z 115 [M+NH4] 1 C00416 LMGP10010000 PA 36:2 718.4 +Neutral Loss Scan m/z 115 [M+NH4] 1 C00416 LMGP10010000 PA 36:3 716.4 +Neutral Loss Scan m/z 115 [M+NH4] 1 C00416 LMGP10010000 PA 36:6 710.4 +Neutral Loss Scan m/z 115 [M+NH4] 1 C00416 LMGP10010000 PA 38:2 746.5 +Neutral Loss Scan m/z 115 [M+NH4] 1 C00416 LMGP10010000 PA 38:3 744.5 +Neutral Loss Scan m/z 115 [M+NH4] 1 C00416 LMGP10010000 PA 38:4 742.5 +Neutral Loss Scan m/z 115 [M+NH4] 1 C00416 LMGP10010000 PA 40:8 762.5 +Neutral Loss Scan m/z 115 [M+NH4] 1 C00416 LMGP10010000 PA 40:9 760.5 +Neutral Loss Scan m/z 115 [M+NH4] 1 C00416 LMGP10010000 METABOLITES_END #END