#METABOLOMICS WORKBENCH Sethjparker_20241101_174730 DATATRACK_ID:5334 STUDY_ID:ST003556 ANALYSIS_ID:AN005846 PROJECT_ID:PR002190
VERSION             	1
CREATED_ON             	November 5, 2024, 3:36 pm
#PROJECT
PR:PROJECT_TITLE                 	N-methyl-arginine significantly inhibits net arginine uptake flux in HEK293
PR:PROJECT_TITLE                 	cells in a dose-dependent manner.
PR:PROJECT_TYPE                  	Manuscript
PR:PROJECT_SUMMARY               	N-methyl-arginine inhibits the metabolism of lysine and arginine by competing
PR:PROJECT_SUMMARY               	for similar transporters. In this project, we aimed to determine the effects of
PR:PROJECT_SUMMARY               	N-methyl-arginine on amino acid uptake flux in HEK293 cells treated for 20-24
PR:PROJECT_SUMMARY               	hours. N-methyl-arginine supplementation inhibited the net uptake of arginine
PR:PROJECT_SUMMARY               	but did not significantly affect lysine or other amino acid uptake fluxes.
PR:INSTITUTE                     	University of British Columbia
PR:DEPARTMENT                    	Biochemistry & Molecular Biology
PR:LABORATORY                    	Parker laboratory
PR:LAST_NAME                     	Parker
PR:FIRST_NAME                    	Seth
PR:ADDRESS                       	950 W 28th Ave, Vancouver, British Columbia, V6H 0B3, Canada
PR:EMAIL                         	seth.parker@bcchr.ca
PR:PHONE                         	6048753121
#STUDY
ST:STUDY_TITLE                   	Metabolomics analysis of conditioned media from N-methyl-arginine-treated HEK293
ST:STUDY_TITLE                   	control (sgTomato) and ALDH7A1-deficient (sgALDH7A1) cells.
ST:STUDY_SUMMARY                 	Metabolite analysis of HEK293 cells treated with N-methyl-arginine (NMA), an
ST:STUDY_SUMMARY                 	inhibitor of lysine/arginine transport. Conditioned media was collected from
ST:STUDY_SUMMARY                 	cells that were treated for 20-24 hours with varying concentration of NMA,
ST:STUDY_SUMMARY                 	centrifuged at 21,000xg to remove cell debris, and 5 ul was added to 250 ul of
ST:STUDY_SUMMARY                 	ice cold 80% methanol containing 1 ul of stable-isotope-labeled internal amino
ST:STUDY_SUMMARY                 	acid standard mix. Metabolites were concentrated using a SpeedVac until dry and
ST:STUDY_SUMMARY                 	analyzed by LCMS. The LC was coupled to an Exploris 240 mass spectrometer
ST:STUDY_SUMMARY                 	operated in a polarity switching data-dependent Top 5 mode. Full MS scan
ST:STUDY_SUMMARY                 	parameters for both positive and negative mode were set to 67-1000 m/z at a
ST:STUDY_SUMMARY                 	resolution of 120k and ddMS2 were collected at a resolution of 30k.
ST:INSTITUTE                     	University of British Columbia
ST:DEPARTMENT                    	Biochemistry & Molecular Biology
ST:LABORATORY                    	Parker laboratory
ST:LAST_NAME                     	Parker
ST:FIRST_NAME                    	Seth
ST:ADDRESS                       	950 W 28th Ave, Vancouver, British Columbia, V6H 0B3, Canada
ST:EMAIL                         	seth.parker@bcchr.ca
ST:PHONE                         	6048753121
#SUBJECT
SU:SUBJECT_TYPE                  	Mammal
SU:SUBJECT_SPECIES               	Homo sapiens
SU:TAXONOMY_ID                   	9606
SU:GENDER                        	Not applicable
#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           	-	Tom_control_CM1	Sample source:Conditioned media | Genotype:CRISPR/Cas9 Control (sgTomato) | Treatment:PBS	Cell Types=HEK293; Independent experiment=1; RAW_FILE_NAME(Raw file name)=Tom_control_CM1.RAW
SUBJECT_SAMPLE_FACTORS           	-	Tom_control_CM2	Sample source:Conditioned media | Genotype:CRISPR/Cas9 Control (sgTomato) | Treatment:PBS	Cell Types=HEK293; Independent experiment=2; RAW_FILE_NAME(Raw file name)=Tom_control_CM2.RAW
SUBJECT_SAMPLE_FACTORS           	-	Tom_control_CM3	Sample source:Conditioned media | Genotype:CRISPR/Cas9 Control (sgTomato) | Treatment:PBS	Cell Types=HEK293; Independent experiment=3; RAW_FILE_NAME(Raw file name)=Tom_control_CM3.RAW
SUBJECT_SAMPLE_FACTORS           	-	Tom_NMA_CM1	Sample source:Conditioned media | Genotype:CRISPR/Cas9 Control (sgTomato) | Treatment:N-methyl-arginine (5 mM)	Cell Types=HEK293; Independent experiment=1; RAW_FILE_NAME(Raw file name)=Tom_NMA_CM1.RAW
SUBJECT_SAMPLE_FACTORS           	-	Tom_NMA_CM2	Sample source:Conditioned media | Genotype:CRISPR/Cas9 Control (sgTomato) | Treatment:N-methyl-arginine (5 mM)	Cell Types=HEK293; Independent experiment=2; RAW_FILE_NAME(Raw file name)=Tom_NMA_CM2.RAW
SUBJECT_SAMPLE_FACTORS           	-	Tom_NMA_CM3	Sample source:Conditioned media | Genotype:CRISPR/Cas9 Control (sgTomato) | Treatment:N-methyl-arginine (5 mM)	Cell Types=HEK293; Independent experiment=3; RAW_FILE_NAME(Raw file name)=Tom_NMA_CM3.RAW
SUBJECT_SAMPLE_FACTORS           	-	KO_control_CM1	Sample source:Conditioned media | Genotype:CRISPR/Cas9 KO (sgALDH7A1) | Treatment:PBS	Cell Types=HEK293; Independent experiment=1; RAW_FILE_NAME(Raw file name)=KO_control_CM1.RAW
SUBJECT_SAMPLE_FACTORS           	-	KO_control_CM2	Sample source:Conditioned media | Genotype:CRISPR/Cas9 KO (sgALDH7A1) | Treatment:PBS	Cell Types=HEK293; Independent experiment=2; RAW_FILE_NAME(Raw file name)=KO_control_CM2.RAW
SUBJECT_SAMPLE_FACTORS           	-	KO_control_CM3	Sample source:Conditioned media | Genotype:CRISPR/Cas9 KO (sgALDH7A1) | Treatment:PBS	Cell Types=HEK293; Independent experiment=3; RAW_FILE_NAME(Raw file name)=KO_control_CM3.RAW
SUBJECT_SAMPLE_FACTORS           	-	KO_NMA_CM1	Sample source:Conditioned media | Genotype:CRISPR/Cas9 KO (sgALDH7A1) | Treatment:N-methyl-arginine (5 mM)	Cell Types=HEK293; Independent experiment=1; RAW_FILE_NAME(Raw file name)=KO_NMA_CM1.RAW
SUBJECT_SAMPLE_FACTORS           	-	KO_NMA_CM2	Sample source:Conditioned media | Genotype:CRISPR/Cas9 KO (sgALDH7A1) | Treatment:N-methyl-arginine (5 mM)	Cell Types=HEK293; Independent experiment=2; RAW_FILE_NAME(Raw file name)=KO_NMA_CM2.RAW
SUBJECT_SAMPLE_FACTORS           	-	KO_NMA_CM3	Sample source:Conditioned media | Genotype:CRISPR/Cas9 KO (sgALDH7A1) | Treatment:N-methyl-arginine (5 mM)	Cell Types=HEK293; Independent experiment=3; RAW_FILE_NAME(Raw file name)=KO_NMA_CM3.RAW
SUBJECT_SAMPLE_FACTORS           	-	HEK293_TOM_0NMA_CM1	Sample source:Conditioned media | Genotype:CRISPR/Cas9 Control (sgTomato) | Treatment:PBS	Cell Types=HEK293; Independent experiment=4; RAW_FILE_NAME(Raw file name)=HEK293_TOM_0NMA_CM1.RAW
SUBJECT_SAMPLE_FACTORS           	-	HEK293_TOM_0NMA_CM2	Sample source:Conditioned media | Genotype:CRISPR/Cas9 Control (sgTomato) | Treatment:PBS	Cell Types=HEK293; Independent experiment=5; RAW_FILE_NAME(Raw file name)=HEK293_TOM_0NMA_CM2.RAW
SUBJECT_SAMPLE_FACTORS           	-	HEK293_TOM_0NMA_CM3	Sample source:Conditioned media | Genotype:CRISPR/Cas9 Control (sgTomato) | Treatment:PBS	Cell Types=HEK293; Independent experiment=6; RAW_FILE_NAME(Raw file name)=HEK293_TOM_0NMA_CM3.RAW
SUBJECT_SAMPLE_FACTORS           	-	HEK293_TOM_5NMA_CM1	Sample source:Conditioned media | Genotype:CRISPR/Cas9 Control (sgTomato) | Treatment:N-methyl-arginine (5 mM)	Cell Types=HEK293; Independent experiment=4; RAW_FILE_NAME(Raw file name)=HEK293_TOM_5NMA_CM1.RAW
SUBJECT_SAMPLE_FACTORS           	-	HEK293_KO_0NMA_CM1	Sample source:Conditioned media | Genotype:CRISPR/Cas9 KO (sgALDH7A1) | Treatment:PBS	Cell Types=HEK293; Independent experiment=4; RAW_FILE_NAME(Raw file name)=HEK293_KO_0NMA_CM1.RAW
SUBJECT_SAMPLE_FACTORS           	-	HEK293_KO_0NMA_CM2	Sample source:Conditioned media | Genotype:CRISPR/Cas9 KO (sgALDH7A1) | Treatment:PBS	Cell Types=HEK293; Independent experiment=5; RAW_FILE_NAME(Raw file name)=HEK293_KO_0NMA_CM2.RAW
SUBJECT_SAMPLE_FACTORS           	-	HEK293_KO_0NMA_CM3	Sample source:Conditioned media | Genotype:CRISPR/Cas9 KO (sgALDH7A1) | Treatment:PBS	Cell Types=HEK293; Independent experiment=6; RAW_FILE_NAME(Raw file name)=HEK293_KO_0NMA_CM3.RAW
SUBJECT_SAMPLE_FACTORS           	-	HEK293_KO_5NMA_CM1	Sample source:Conditioned media | Genotype:CRISPR/Cas9 KO (sgALDH7A1) | Treatment:N-methyl-arginine (5 mM)	Cell Types=HEK293; Independent experiment=4; RAW_FILE_NAME(Raw file name)=HEK293_KO_5NMA_CM1.RAW
SUBJECT_SAMPLE_FACTORS           	-	HEK293_TOM_10NMA_CM1	Sample source:Conditioned media | Genotype:CRISPR/Cas9 Control (sgTomato) | Treatment:N-methyl-arginine (10 mM)	Cell Types=HEK293; Independent experiment=4; RAW_FILE_NAME(Raw file name)=HEK293_TOM_10NMA_CM1.RAW
SUBJECT_SAMPLE_FACTORS           	-	HEK293_TOM_10NMA_CM2	Sample source:Conditioned media | Genotype:CRISPR/Cas9 Control (sgTomato) | Treatment:N-methyl-arginine (10 mM)	Cell Types=HEK293; Independent experiment=5; RAW_FILE_NAME(Raw file name)=HEK293_TOM_10NMA_CM2.RAW
SUBJECT_SAMPLE_FACTORS           	-	HEK293_TOM_10NMA_CM3	Sample source:Conditioned media | Genotype:CRISPR/Cas9 Control (sgTomato) | Treatment:N-methyl-arginine (10 mM)	Cell Types=HEK293; Independent experiment=6; RAW_FILE_NAME(Raw file name)=HEK293_TOM_10NMA_CM3.RAW
SUBJECT_SAMPLE_FACTORS           	-	HEK293_TOM_20NMA_CM1	Sample source:Conditioned media | Genotype:CRISPR/Cas9 Control (sgTomato) | Treatment:N-methyl-arginine (20 mM)	Cell Types=HEK293; Independent experiment=4; RAW_FILE_NAME(Raw file name)=HEK293_TOM_20NMA_CM1.RAW
SUBJECT_SAMPLE_FACTORS           	-	HEK293_TOM_20NMA_CM2	Sample source:Conditioned media | Genotype:CRISPR/Cas9 Control (sgTomato) | Treatment:N-methyl-arginine (20 mM)	Cell Types=HEK293; Independent experiment=5; RAW_FILE_NAME(Raw file name)=HEK293_TOM_20NMA_CM2.RAW
SUBJECT_SAMPLE_FACTORS           	-	HEK293_TOM_20NMA_CM3	Sample source:Conditioned media | Genotype:CRISPR/Cas9 Control (sgTomato) | Treatment:N-methyl-arginine (20 mM)	Cell Types=HEK293; Independent experiment=6; RAW_FILE_NAME(Raw file name)=HEK293_TOM_20NMA_CM3.RAW
SUBJECT_SAMPLE_FACTORS           	-	HEK293_KO_10NMA_CM1	Sample source:Conditioned media | Genotype:CRISPR/Cas9 KO (sgALDH7A1) | Treatment:N-methyl-arginine (10 mM)	Cell Types=HEK293; Independent experiment=4; RAW_FILE_NAME(Raw file name)=HEK293_KO_10NMA_CM1.RAW
SUBJECT_SAMPLE_FACTORS           	-	HEK293_KO_10NMA_CM2	Sample source:Conditioned media | Genotype:CRISPR/Cas9 KO (sgALDH7A1) | Treatment:N-methyl-arginine (10 mM)	Cell Types=HEK293; Independent experiment=5; RAW_FILE_NAME(Raw file name)=HEK293_KO_10NMA_CM2.RAW
SUBJECT_SAMPLE_FACTORS           	-	HEK293_KO_10NMA_CM3	Sample source:Conditioned media | Genotype:CRISPR/Cas9 KO (sgALDH7A1) | Treatment:N-methyl-arginine (10 mM)	Cell Types=HEK293; Independent experiment=6; RAW_FILE_NAME(Raw file name)=HEK293_KO_10NMA_CM3.RAW
SUBJECT_SAMPLE_FACTORS           	-	HEK293_KO_20NMA_CM1	Sample source:Conditioned media | Genotype:CRISPR/Cas9 KO (sgALDH7A1) | Treatment:N-methyl-arginine (20 mM)	Cell Types=HEK293; Independent experiment=4; RAW_FILE_NAME(Raw file name)=HEK293_KO_20NMA_CM1.RAW
SUBJECT_SAMPLE_FACTORS           	-	HEK293_KO_20NMA_CM2	Sample source:Conditioned media | Genotype:CRISPR/Cas9 KO (sgALDH7A1) | Treatment:N-methyl-arginine (20 mM)	Cell Types=HEK293; Independent experiment=5; RAW_FILE_NAME(Raw file name)=HEK293_KO_20NMA_CM2.RAW
SUBJECT_SAMPLE_FACTORS           	-	HEK293_KO_20NMA_CM3	Sample source:Conditioned media | Genotype:CRISPR/Cas9 KO (sgALDH7A1) | Treatment:N-methyl-arginine (20 mM)	Cell Types=HEK293; Independent experiment=6; RAW_FILE_NAME(Raw file name)=HEK293_KO_20NMA_CM3.RAW
SUBJECT_SAMPLE_FACTORS           	-	CMt0_control_CM1	Sample source:Conditioned media | Genotype:None | Treatment:PBS	Cell Types=None; Independent experiment=1; RAW_FILE_NAME(Raw file name)=CMt0_control_CM1.RAW
SUBJECT_SAMPLE_FACTORS           	-	DMEMto_control_CM2	Sample source:Conditioned media | Genotype:None | Treatment:PBS	Cell Types=None; Independent experiment=2; RAW_FILE_NAME(Raw file name)=DMEMto_control_CM2.RAW
SUBJECT_SAMPLE_FACTORS           	-	DMEMto_control_CM3	Sample source:Conditioned media | Genotype:None | Treatment:PBS	Cell Types=None; Independent experiment=3; RAW_FILE_NAME(Raw file name)=DMEMto_control_CM3.RAW
SUBJECT_SAMPLE_FACTORS           	-	CMt0_NMA_CM1	Sample source:Conditioned media | Genotype:None | Treatment:N-methyl-arginine (5 mM)	Cell Types=None; Independent experiment=1; RAW_FILE_NAME(Raw file name)=CMt0_NMA_CM1.RAW
SUBJECT_SAMPLE_FACTORS           	-	DMEMto_NMA_CM2	Sample source:Conditioned media | Genotype:None | Treatment:N-methyl-arginine (5 mM)	Cell Types=None; Independent experiment=2; RAW_FILE_NAME(Raw file name)=DMEMto_NMA_CM2.RAW
SUBJECT_SAMPLE_FACTORS           	-	DMEMto_NMA_CM3	Sample source:Conditioned media | Genotype:None | Treatment:N-methyl-arginine (5 mM)	Cell Types=None; Independent experiment=3; RAW_FILE_NAME(Raw file name)=DMEMto_NMA_CM3.RAW
SUBJECT_SAMPLE_FACTORS           	-	DMEMt0_0NMA_CM1	Sample source:Conditioned media | Genotype:None | Treatment:PBS	Cell Types=None; Independent experiment=4; RAW_FILE_NAME(Raw file name)=DMEMt0_0NMA_CM1.RAW
SUBJECT_SAMPLE_FACTORS           	-	DMEMt0_0NMA_CM2	Sample source:Conditioned media | Genotype:None | Treatment:PBS	Cell Types=None; Independent experiment=5; RAW_FILE_NAME(Raw file name)=DMEMt0_0NMA_CM2.RAW
SUBJECT_SAMPLE_FACTORS           	-	DMEMt0_0NMA_CM3	Sample source:Conditioned media | Genotype:None | Treatment:PBS	Cell Types=None; Independent experiment=6; RAW_FILE_NAME(Raw file name)=DMEMt0_0NMA_CM3.RAW
SUBJECT_SAMPLE_FACTORS           	-	DMEMt0_5NMA_CM1	Sample source:Conditioned media | Genotype:None | Treatment:N-methyl-arginine (5 mM)	Cell Types=None; Independent experiment=4; RAW_FILE_NAME(Raw file name)=DMEMt0_5NMA_CM1.RAW
SUBJECT_SAMPLE_FACTORS           	-	DMEMt0_10NMA_CM1	Sample source:Conditioned media | Genotype:None | Treatment:N-methyl-arginine (10 mM)	Cell Types=None; Independent experiment=4; RAW_FILE_NAME(Raw file name)=DMEMt0_10NMA_CM1.RAW
SUBJECT_SAMPLE_FACTORS           	-	DMEMt0_10NMA_CM2	Sample source:Conditioned media | Genotype:None | Treatment:N-methyl-arginine (10 mM)	Cell Types=None; Independent experiment=5; RAW_FILE_NAME(Raw file name)=DMEMt0_10NMA_CM2.RAW
SUBJECT_SAMPLE_FACTORS           	-	DMEMt0_10NMA_CM3	Sample source:Conditioned media | Genotype:None | Treatment:N-methyl-arginine (10 mM)	Cell Types=None; Independent experiment=6; RAW_FILE_NAME(Raw file name)=DMEMt0_10NMA_CM3.RAW
SUBJECT_SAMPLE_FACTORS           	-	DMEMt0_20NMA_CM1	Sample source:Conditioned media | Genotype:None | Treatment:N-methyl-arginine (20 mM)	Cell Types=None; Independent experiment=4; RAW_FILE_NAME(Raw file name)=DMEMt0_20NMA_CM1.RAW
SUBJECT_SAMPLE_FACTORS           	-	DMEMt0_20NMA_CM2	Sample source:Conditioned media | Genotype:None | Treatment:N-methyl-arginine (20 mM)	Cell Types=None; Independent experiment=5; RAW_FILE_NAME(Raw file name)=DMEMt0_20NMA_CM2.RAW
SUBJECT_SAMPLE_FACTORS           	-	DMEMt0_20NMA_CM3	Sample source:Conditioned media | Genotype:None | Treatment:N-methyl-arginine (20 mM)	Cell Types=None; Independent experiment=6; RAW_FILE_NAME(Raw file name)=DMEMt0_20NMA_CM3.RAW
#COLLECTION
CO:COLLECTION_SUMMARY            	5 ul of initial or conditioned media was added to 250 ul of ice cold 80%
CO:COLLECTION_SUMMARY            	methanol containing isotopic amino acid internal standards.
CO:SAMPLE_TYPE                   	Media
#TREATMENT
TR:TREATMENT_SUMMARY             	Cells were plated in DMEM and allowed to attach overnight. The following
TR:TREATMENT_SUMMARY             	morning, media was aspirated and replaced with DMEM containing 10% dialyzed FBS
TR:TREATMENT_SUMMARY             	and a vehicle (PBS) or N-methyl-arginine ranging from 5-20 mM. Media was
TR:TREATMENT_SUMMARY             	collected and centrifuged at 1,000xg to remove cell debris. 5 ul of conditioned
TR:TREATMENT_SUMMARY             	or initial media was measured using LCMS to quantify changes in amino acid
TR:TREATMENT_SUMMARY             	levels.
TR:TREATMENT_COMPOUND            	N-methyl-arginine
TR:TREATMENT_DOSE                	5-20 mM
TR:TREATMENT_VEHICLE             	PBS
#SAMPLEPREP
SP:SAMPLEPREP_SUMMARY            	Dried samples were reconstituted in 50 µL of HPLC-grade water. Samples were
SP:SAMPLEPREP_SUMMARY            	vortexed for ~10 minutes, then centrifuged at 21,000 x g for 15 min at 4°C. 40
SP:SAMPLEPREP_SUMMARY            	microliters were transferred to LC vials containing glass inserts for analysis.
#CHROMATOGRAPHY
CH:CHROMATOGRAPHY_TYPE           	HILIC
CH:INSTRUMENT_NAME               	Thermo Vanquish
CH:COLUMN_NAME                   	Merck SeQuant ZIC-pHILIC (150 x 2.1mm,5um)
CH:SOLVENT_A                     	100% water, 10 mM Ammonium Carbonate, pH 9.0
CH:SOLVENT_B                     	100% acetonitrile
CH:FLOW_GRADIENT                 	80-20%B (0-30 min), 20-20%B (30-40 minute), and 20-80%B (40-40.5 minute); the LC
CH:FLOW_GRADIENT                 	column was re-equilibrated using 80-80%B from 40.5-52 minute before subsequent
CH:FLOW_GRADIENT                 	injections
CH:FLOW_RATE                     	100 uL/min
CH:COLUMN_TEMPERATURE            	25
#ANALYSIS
AN:ANALYSIS_TYPE                 	MS
#MS
MS:INSTRUMENT_NAME               	Thermo Exploris 240
MS:INSTRUMENT_TYPE               	Orbitrap
MS:MS_TYPE                       	ESI
MS:ION_MODE                      	NEGATIVE
MS:MS_COMMENTS                   	The LC was coupled to a Thermo Scientific Exploris 240 mass spectrometer
MS:MS_COMMENTS                   	operating in heated electrospray ionization mode (HESI) for analysis. The
MS:MS_COMMENTS                   	following parameters were set for HESI: spray voltage 3.4 kV (positive) and 2 kV
MS:MS_COMMENTS                   	(negative), static spray voltage, sheath gas 25, aux gas 5, sweep gas 0.5, ion
MS:MS_COMMENTS                   	transfer tube temperature 320oC, and vaporizer temperature 75oC. The global
MS:MS_COMMENTS                   	parameters included an expected peak width of 20 seconds, mild trapping, and a
MS:MS_COMMENTS                   	default charge state of 1. A 40-min polarity switching data-dependent Top 5
MS:MS_COMMENTS                   	method was used for positive mode and a data-dependent Top 3 method was used for
MS:MS_COMMENTS                   	negative mode. Full MS scan parameters for both positive and negative modes were
MS:MS_COMMENTS                   	set as follows: scan range 67-1000 m/z collected in profile mode, Orbitrap
MS:MS_COMMENTS                   	resolution 120,000, RF lens 70%, AGC target of 300%, and maximum injection time
MS:MS_COMMENTS                   	set to automatic. ddMS2 for positive mode were collected in centroid mode at an
MS:MS_COMMENTS                   	Orbitrap resolution of 30,000, isolation window of 1.5 m/z, an AGC target set to
MS:MS_COMMENTS                   	standard, a maximum injection time set to automatic, and a normalized collision
MS:MS_COMMENTS                   	energy set to 10%, 30%, and 80%. ddMS2 for negative mode were collected in
MS:MS_COMMENTS                   	centroid mode at an Orbitrap resolution of 30,000, isolation window of 2 m/z, an
MS:MS_COMMENTS                   	AGC target set to standard, a maximum injection time set to automatic, and a
MS:MS_COMMENTS                   	normalized collision energy set to 30%. For both positive and negative ddMS2, we
MS:MS_COMMENTS                   	applied an intensity threshold of 5e4 and a dynamic exclusion of 5 ppm for 10
MS:MS_COMMENTS                   	seconds, excluding isotopes. A targeted selected ion monitoring (tSIM) scan was
MS:MS_COMMENTS                   	also included for pipecolate and P6C/P2C, and the retention time ranges were
MS:MS_COMMENTS                   	based on elution of authentic standards (pipecolate) or from positive samples
MS:MS_COMMENTS                   	(Aldh7a1-deficient tissues). The tSIM scan for pipecolate was collected from
MS:MS_COMMENTS                   	8-12 minutes in negative mode at an isolation window of 4 m/z (for metabolomics)
MS:MS_COMMENTS                   	or 18 m/z (for isotope-tracing experiments, to include m/z shifts), an Orbitrap
MS:MS_COMMENTS                   	resolution of 120,000, a RF lens at 70%, an automatic maximum injection time,
MS:MS_COMMENTS                   	and collected in profile mode. The tSIM scan for P6C/P2C was collected from 6-10
MS:MS_COMMENTS                   	minutes in positive mode at an isolation window of 4 m/z (for metabolomics) or
MS:MS_COMMENTS                   	18 m/z (for isotope-tracing experiments, to include m/z shifts), an Orbitrap
MS:MS_COMMENTS                   	resolution of 120,000, a RF lens at 70%, an automatic maximum injection time,
MS:MS_COMMENTS                   	and collected in profile mode.
#MS_METABOLITE_DATA
MS_METABOLITE_DATA:UNITS	ion counts
MS_METABOLITE_DATA_START
Samples	CMt0_control_CM1	Tom_control_CM1	KO_control_CM1	CMt0_NMA_CM1	Tom_NMA_CM1	KO_NMA_CM1	DMEMto_control_CM2	Tom_control_CM2	KO_control_CM2	DMEMto_NMA_CM2	Tom_NMA_CM2	KO_NMA_CM2	DMEMto_control_CM3	Tom_control_CM3	KO_control_CM3	DMEMto_NMA_CM3	Tom_NMA_CM3	KO_NMA_CM3	DMEMt0_0NMA_CM1	DMEMt0_0NMA_CM2	DMEMt0_0NMA_CM3	DMEMt0_10NMA_CM1	DMEMt0_10NMA_CM2	DMEMt0_10NMA_CM3	DMEMt0_20NMA_CM1	DMEMt0_20NMA_CM2	DMEMt0_20NMA_CM3	DMEMt0_5NMA_CM1	HEK293_KO_0NMA_CM1	HEK293_KO_0NMA_CM2	HEK293_KO_0NMA_CM3	HEK293_KO_10NMA_CM1	HEK293_KO_10NMA_CM2	HEK293_KO_10NMA_CM3	HEK293_KO_20NMA_CM1	HEK293_KO_20NMA_CM2	HEK293_KO_20NMA_CM3	HEK293_KO_5NMA_CM1	HEK293_TOM_0NMA_CM1	HEK293_TOM_0NMA_CM2	HEK293_TOM_0NMA_CM3	HEK293_TOM_10NMA_CM1	HEK293_TOM_10NMA_CM2	HEK293_TOM_10NMA_CM3	HEK293_TOM_20NMA_CM1	HEK293_TOM_20NMA_CM2	HEK293_TOM_20NMA_CM3	HEK293_TOM_5NMA_CM1
Factors	Sample source:Conditioned media | Genotype:None | Treatment:PBS	Sample source:Conditioned media | Genotype:CRISPR/Cas9 Control (sgTomato) | Treatment:PBS	Sample source:Conditioned media | Genotype:CRISPR/Cas9 KO (sgALDH7A1) | Treatment:PBS	Sample source:Conditioned media | Genotype:None | Treatment:N-methyl-arginine (5 mM)	Sample source:Conditioned media | Genotype:CRISPR/Cas9 Control (sgTomato) | Treatment:N-methyl-arginine (5 mM)	Sample source:Conditioned media | Genotype:CRISPR/Cas9 KO (sgALDH7A1) | Treatment:N-methyl-arginine (5 mM)	Sample source:Conditioned media | Genotype:None | Treatment:PBS	Sample source:Conditioned media | Genotype:CRISPR/Cas9 Control (sgTomato) | Treatment:PBS	Sample source:Conditioned media | Genotype:CRISPR/Cas9 KO (sgALDH7A1) | Treatment:PBS	Sample source:Conditioned media | Genotype:None | Treatment:N-methyl-arginine (5 mM)	Sample source:Conditioned media | Genotype:CRISPR/Cas9 Control (sgTomato) | Treatment:N-methyl-arginine (5 mM)	Sample source:Conditioned media | Genotype:CRISPR/Cas9 KO (sgALDH7A1) | Treatment:N-methyl-arginine (5 mM)	Sample source:Conditioned media | Genotype:None | Treatment:PBS	Sample source:Conditioned media | Genotype:CRISPR/Cas9 Control (sgTomato) | Treatment:PBS	Sample source:Conditioned media | Genotype:CRISPR/Cas9 KO (sgALDH7A1) | Treatment:PBS	Sample source:Conditioned media | Genotype:None | Treatment:N-methyl-arginine (5 mM)	Sample source:Conditioned media | Genotype:CRISPR/Cas9 Control (sgTomato) | Treatment:N-methyl-arginine (5 mM)	Sample source:Conditioned media | Genotype:CRISPR/Cas9 KO (sgALDH7A1) | Treatment:N-methyl-arginine (5 mM)	Sample source:Conditioned media | Genotype:None | Treatment:PBS	Sample source:Conditioned media | Genotype:None | Treatment:PBS	Sample source:Conditioned media | Genotype:None | Treatment:PBS	Sample source:Conditioned media | Genotype:None | Treatment:N-methyl-arginine (10 mM)	Sample source:Conditioned media | Genotype:None | Treatment:N-methyl-arginine (10 mM)	Sample source:Conditioned media | Genotype:None | Treatment:N-methyl-arginine (10 mM)	Sample source:Conditioned media | Genotype:None | Treatment:N-methyl-arginine (20 mM)	Sample source:Conditioned media | Genotype:None | Treatment:N-methyl-arginine (20 mM)	Sample source:Conditioned media | Genotype:None | Treatment:N-methyl-arginine (20 mM)	Sample source:Conditioned media | Genotype:None | Treatment:N-methyl-arginine (5 mM)	Sample source:Conditioned media | Genotype:CRISPR/Cas9 KO (sgALDH7A1) | Treatment:PBS	Sample source:Conditioned media | Genotype:CRISPR/Cas9 KO (sgALDH7A1) | Treatment:PBS	Sample source:Conditioned media | Genotype:CRISPR/Cas9 KO (sgALDH7A1) | Treatment:PBS	Sample source:Conditioned media | Genotype:CRISPR/Cas9 KO (sgALDH7A1) | Treatment:N-methyl-arginine (10 mM)	Sample source:Conditioned media | Genotype:CRISPR/Cas9 KO (sgALDH7A1) | Treatment:N-methyl-arginine (10 mM)	Sample source:Conditioned media | Genotype:CRISPR/Cas9 KO (sgALDH7A1) | Treatment:N-methyl-arginine (10 mM)	Sample source:Conditioned media | Genotype:CRISPR/Cas9 KO (sgALDH7A1) | Treatment:N-methyl-arginine (20 mM)	Sample source:Conditioned media | Genotype:CRISPR/Cas9 KO (sgALDH7A1) | Treatment:N-methyl-arginine (20 mM)	Sample source:Conditioned media | Genotype:CRISPR/Cas9 KO (sgALDH7A1) | Treatment:N-methyl-arginine (20 mM)	Sample source:Conditioned media | Genotype:CRISPR/Cas9 KO (sgALDH7A1) | Treatment:N-methyl-arginine (5 mM)	Sample source:Conditioned media | Genotype:CRISPR/Cas9 Control (sgTomato) | Treatment:PBS	Sample source:Conditioned media | Genotype:CRISPR/Cas9 Control (sgTomato) | Treatment:PBS	Sample source:Conditioned media | Genotype:CRISPR/Cas9 Control (sgTomato) | Treatment:PBS	Sample source:Conditioned media | Genotype:CRISPR/Cas9 Control (sgTomato) | Treatment:N-methyl-arginine (10 mM)	Sample source:Conditioned media | Genotype:CRISPR/Cas9 Control (sgTomato) | Treatment:N-methyl-arginine (10 mM)	Sample source:Conditioned media | Genotype:CRISPR/Cas9 Control (sgTomato) | Treatment:N-methyl-arginine (10 mM)	Sample source:Conditioned media | Genotype:CRISPR/Cas9 Control (sgTomato) | Treatment:N-methyl-arginine (20 mM)	Sample source:Conditioned media | Genotype:CRISPR/Cas9 Control (sgTomato) | Treatment:N-methyl-arginine (20 mM)	Sample source:Conditioned media | Genotype:CRISPR/Cas9 Control (sgTomato) | Treatment:N-methyl-arginine (20 mM)	Sample source:Conditioned media | Genotype:CRISPR/Cas9 Control (sgTomato) | Treatment:N-methyl-arginine (5 mM)
ALANINE / SARCOSINE	1912432.206	142568807.1	156145877.9	1380953.861	130752207.4	108772585.5	491806.2426	117680159.8	100007832	331421.4837	110003426.9	98646306.04	63523994.44	161157773.6	164047126.9	65803807.54	146034805	152867235.7	3145078.855	1922095.425	4152468.012	2517430.048	1865799.829	2138095.627	2009167.779	2036702.674	3424901.166	2081245.326	86303434.51	117544663.8	78903076.77	67573953.09	91800259.9	101638036.1	82280311.68	78213951.23	86966880.19	80581254.95	75980512.31	118632717.6	120253847.6	67421442.59	87757292.22	81089304.18	60184112.17	73138513.89	33983623.53	60319401.38
iso_Ala	201083782	185692550.4	173426595.7	195479062.8	182412268.3	132160002.8	83270305.71	171493975.5	181007096.4	60764978.71	172048612.2	179216948.9	161733188.3	166016674.4	164488823.6	161551795.8	160850990.9	161715090.2	174553755.1	191643460.4	195934588.5	173783202.9	183632435	196641532.3	169535802.1	188378231.7	186200123.5	186107430.5	168190975.4	173197865.7	102416669.3	179838985.3	184853521.3	179781484.2	178269243.7	187297976.8	186116133.2	182281318.1	141539941.5	175980498.6	186444740.1	166148003.4	176863848.1	183421163.5	181974283.1	183880542	90668122.22	125985959.8
ARGININE	1176116454	905123497.5	897202772.8	1073343020	875249712.4	665159255.3	475774402.1	754259471.4	684100962.6	354383761.1	837708787.6	747015049.8	886264416.5	737174238.8	709756073.9	851273799.4	709931764.9	686533748.9	805660866.7	970549644.9	1049386077	733613172.8	865936256.4	970230916.3	696322244.9	865548472.5	905853885.6	776900427.2	714417709.4	731152830.1	444836837.9	718411391	772983249.2	768102098.9	745697875.4	762202968.7	829849946.1	738033712.8	556064706.1	734699157.6	822003965.2	677662186.1	738287700.9	801475915	738071307.5	760812577.6	388049842.7	474376331.6
iso_Arg	697716766.7	726728626.6	708020630.6	631743968.1	627761046.8	488952629.3	346989291.7	733063610.4	780707587.8	247185976.2	688339844.5	705331375.8	566988348.2	616323376.4	603894002.6	526664769.7	532213095.2	541590387.4	573002593	653095368.7	720552009.2	509411939.5	584439448	658044598.6	485122379.4	564579059.3	600022262.2	546896124.7	583037110.8	655792400.8	412669172.4	565698696.3	618733503.6	630454464.7	545113647.1	582949201.5	623893663.3	578437949.5	485827745.9	668859455.8	726252688	529058477.8	602275311.1	639943510.6	543360035.8	571653069.3	297579501.5	394836876.1
ASPARAGINE	225081.4925	6637329.638	8412651.275	154788.0386	6173152.776	6294839.903	64810.28368	7454180.499	5352816.505	0	9433385.05	5987441.459	1742923.108	7079123.142	7357205.967	1616791.692	6069220.641	6644539.871	200700.5541	106560.0438	144628.5435	87603.27787	87860.25055	91829.52461	0	69640.76341	148033.8898	76826.25355	4884736.695	7678895.785	4321409.282	4617134.901	6797570.955	6424680.91	5472927.31	8004001.082	7703332.068	4867362.521	4323331.747	6051604.924	5666553.134	4844871.105	5552072.614	4972792.827	6097586.304	6808428.971	2962856.761	3431311.128
iso_Asn	79966332.39	80580192.09	77194669.49	77949282.99	80043255.35	60796019.73	32941502.86	75322757.85	78344123.68	24459296.63	78304269.04	79259730.8	70582249.13	77610096.36	75550118.39	70690821.26	76119353.73	75391791.23	67711501.78	81722266.07	84299961.3	69939180.42	79748184.06	87237899.39	72151413.35	83689470.57	86213210.85	72254065.49	70378749.4	83753660.07	47313313.28	76691799.51	86406548.42	83517038.79	80443612.12	87810814.41	88824944.37	74786564.88	61422657.63	82172708.51	83593316.21	72890162.41	83183343.57	84666353.63	79188331.09	85324494.84	41785419.07	51949434.83
ASPARTATE	566591.3451	753608.8014	337780.9696	407696.2881	189185.5287	421000.33	151955.0791	482079.8712	329323.3428	106924.0573	672639.324	245737.0336	4995623.013	654762.6191	489362.9073	5042934.885	895055.4657	712340.2617	1085330.569	787896.1154	963515.0973	546021.1038	311762.9847	623405.4207	500320.5409	621984.3375	1042133.732	713968.3674	1068525.697	419195.5884	307563.5629	1469759.546	1002386.099	1238163.634	1078791.568	1110331.798	513958.3883	1216799.53	380272.243	606422.6623	519847.6186	461303.0064	672143.4083	672783.2901	379754.3479	921959.0303	304828.9931	665557.5816
iso_Asp	116879079.9	117151315.9	114383706.8	117338717.7	114278896.5	83421989	49343021.3	109348415.9	117790408.8	35832589.03	111735569.7	118172856.7	110985364	116191636.6	115306985.4	108656553.3	114763920	116912421.9	120817623.2	113671395.5	115647835.6	111286947.2	112878452.8	115874034	102618482.4	115122832.5	111768429.4	113878000.8	116836208.2	116355941.6	62953106.19	119563325.4	117054110.5	110626354.9	116263719.2	116131004.8	112735804.9	115520034.6	98534196.91	113142228	111690885	108674054.5	115781756.9	111824758.4	112944903.7	116137521.7	50484206.82	82778722.11
CYSTINE	238028321.1	203100005.6	186942104	232300790.5	192033275.5	132248477.8	63084118.34	145261880.9	119938016.7	45541866.59	156901125.1	127822869.2	200004111.7	198948678	182298651.3	172426909.9	174286455.7	160582332.9	149611977.8	278653918	299110437.7	137303989.2	246803909.8	284076978	143973280.9	259547735.7	260770467.5	146706894.3	143098829.8	218631849.2	116776661.7	128379773.3	225626990.1	199698274.4	153527717.4	223608168.4	223924584.5	136537297.4	115426827.4	235333906.8	253045790.7	136231261.3	210028353.1	227542585.6	152583716	213025576.3	98333437	85506722.65
iso_CysCys	140247303.9	133653890.7	125852296.8	133999441.2	125458568	92589708.37	47139081.32	117362957	126839729	32779468.85	118803679.3	117854025.8	123390760.8	134965046.5	128270663.1	104248591.8	116848503	112548526	92361230.56	159539137	173327068.2	82191423.6	139921372.8	157976770.4	82870986.52	141060241.7	143550413.5	86991073.32	89685097.45	142371909.8	76003513.39	83649656.55	144454310.4	128210047.3	95642115.5	146107211.8	139592935	88776665.62	75454468.88	146980813.6	155902206.7	85984606.55	133392958.1	140594701.3	95949858.48	134123909.8	60980245.75	56362816.59
GLUTAMINE	3534371984	2890756896	2867774694	3463250557	2992909960	2212736787	1416827819	2404715048	2209244561	1113632185	2626775787	2397159931	2965641430	2745208491	2727126064	3035711113	2787713362	2735184653	2889923074	3094072962	3149348157	2748224529	3021028430	3199548026	2682917445	3117873989	3174715819	2793408966	2654079244	2648580620	1512119502	2656320275	2758596880	2672903652	2746946184	2753638474	2815732915	2708803239	2094980336	2615092857	2660810156	2531090377	2631305002	2736328603	2676157042	2786217622	1414296391	1810274443
iso_Gln	252526047.8	264437118.6	257178308.9	247628428.6	266290502	199405542.4	122078373.1	262858738.6	284327766.4	92557849.83	269101631.2	278953857.2	224353684.8	246243934.9	248082527.7	223933889	245402464.1	246633412.5	228408354.4	234252219.4	238856942.4	215443948.4	228746569.7	240201105.1	212844878.5	230136298.3	234665444.4	223213665	229976911.9	251094580.1	145529202.3	243135030.4	253226394.5	247884483.3	236520103	249569566.2	249732350.1	242404733.2	199557777.3	246944572.6	243933327.7	230508701.1	245152635.1	245385580.6	233881867.9	247484298.7	126366105.7	173489247.3
GLUTAMATE	14820160.12	10045281.2	7252539.066	13885958.02	4998996.851	6996321.337	5367455.113	4824896.977	5151520.75	3929460.822	5812204.657	5892162.954	96516053.17	12206628.59	8529758.213	107950164.2	21214322.06	17146967.29	25431847.49	18469818.89	20093968.79	14518824.73	17763670.03	20660652.17	13741328.02	18167920.67	24467168.87	15306167.84	16180870.16	7755405.543	3953196.402	20931294.88	16430106.1	23424478.47	12561962.86	20139058.49	10443342.36	19597235.8	5008955.573	6550111.046	8226761.26	5898387.505	8855849.985	12284911.7	7868532.113	14121973.38	3687732.803	8184579.114
iso_Glu	302861191.6	296328535.3	283958624.8	301465993.1	288363506.8	218063141.2	118626257.4	269332107.9	284061969	84579239.21	272276349.4	280981179.8	252214826.4	280830884.1	278160074.1	247243871.9	269160299.2	270808400.2	269514990.1	281687472.3	300332822.4	259159462.9	277316491	302919871.1	254125100.5	284996639.2	290240327.8	279869520.5	272066339.1	280254807.4	167449826.9	277482679.5	288831031.5	283065108.3	279210446.6	283568464.7	294738608.6	280372357.6	233115890.7	282129686.9	297095779.6	259823269.4	278918266.6	290897487.8	277891560.4	284400849.1	138941146.2	200187083.6
GLYCINE	58211506.54	28570736.65	25927507.1	25563461.37	26169642.3	27920872.81	6172901.481	20755884.55	18750672.92	4569460.315	24105730.2	20316728.84	21442443.75	25889067.74	25611053.1	42279963.92	25422753.37	25657293.42	58150202.59	30402063.92	29298532.52	24246473.54	27167469.79	31367752.09	23031044.84	28748137.72	32834358.13	37008463.98	28117652.82	30853879.16	13889572.54	33134119.38	31333967.29	27919537.38	29459483.97	32934210.99	33562841.8	38111114.64	20882509.32	30195992.66	31009838.15	28187892.25	28168275.22	34386828.53	32662960.27	32759654.46	10985613.83	17512046.21
iso_Gly	21562524.87	21371314.98	19989469.86	20701633.52	19593763.21	13992362.46	6525293.425	19750122.86	19875428.74	4823084.238	19782375.5	20019664.7	16254076.82	18645925.56	18406435.13	15943820.55	16972716.21	17306295.07	20751276.34	23535135.29	22943355.48	19566480	21369594.57	23581962.6	19507034.05	22569622.18	21189744.6	20868043.64	20626057.13	22682369.32	10489068.51	20844639.51	23694977.42	19714767.99	21903577.25	23442256.9	23317370.35	21725092.01	16644498.86	22678284.09	22884271.57	20511768.47	21914564.75	21249040.05	23484632.49	22333537.29	8725347.31	13679828.34
LYSINE	356763811	320484067.2	314071496	254303987.4	229808006.5	173492527.9	104111541.5	239609611.7	212455424.6	59589169.7	196568740.5	172685900.1	291728267.8	289842131.4	275526003.9	227863733.3	216125361.8	203182029.3	268907005.4	342217164.3	366465713.9	203343086	234721793	257578665	186425515.8	229510971.1	237417357.1	222933808.5	261469063	276815199.6	161535197.1	208295489.2	226285682.4	223117143.2	206013698.3	211270867	219311691.6	223461039.3	200352934.3	304599037.8	330956989.5	194361742.9	217412585.3	230438467.4	199145266.5	209571113.2	97663869.86	141443299.5
iso_Lys	94595307.27	96480396.75	92439401.57	67741580.68	67535914.88	51372506.27	32881077.54	83227704.17	88398871.97	18332578.04	63290659.87	65458017.91	73571139.09	79829672.42	77261796.44	56724702.22	59084705.62	57368506.49	82838641.06	98082888.46	106059363.8	62585352.78	68356589.53	75090052.1	58092882.26	65477533.88	67567025.62	70138100.48	82061623.4	88686507.38	50818990.42	67870229.66	71976630.04	69766938.09	64490167.22	66987971.57	67451883.17	72500253.79	66343410.85	96343952.22	101703097.2	62688142.82	69486240.79	71229339.64	62763602.02	64502547.93	30386315.49	47355309.64
METHIONINE	265344037.5	220112252.4	222102927.7	259150980.2	224454869.6	158275889.4	84767998	172135374.6	156939312.5	62806574.84	191828356.7	171056517.9	216360114.6	208086850.4	197241033.4	223086245.4	204856324.9	196023740.4	193912569.7	234149972.4	238798668.3	198419997.5	231430892	245717707.4	190852635.8	240593266.5	242758024.8	207574123.3	195625037.6	202393262.3	108209652.7	201595823	216312884	207251558	211622728.5	217345016.5	220955919.7	207497207.9	153680978.5	205510589.9	210465958.1	187065861.4	203754614.2	213758632.6	205721183.6	216902594.9	93561129.71	130204120.6
iso_Met	306231555.5	304732056.9	298311709.9	298218308.9	304145578.9	213705077.5	115877659.3	282476539.3	299240525	83250279.49	291824995	296732158.6	252265266.5	279422219.3	270898569.5	248829555.4	267833390.4	267940611.5	243616576.7	269867711	278032642.9	243123968.4	268492291.4	283465859	237387671.1	272379319.6	275061796	258207133.2	256013628.8	273139413.1	144276765.4	272120655.9	286728137.6	278072680.1	274363949	286125994.1	284013002.8	275206824.7	213593605.3	274559965	276733826.7	251332776.9	275033224.4	280412972.7	269882141.3	280034588.2	120882904.1	181696653.6
PHENYLALANINE	1198550110	1028645284	1043818594	1163189446	1068780480	730918342.5	353441457.8	819894972.7	735768470.3	261888276	903243577.5	795948579.7	949154929.3	988523164.1	945335838.6	987333426.2	962293677.4	910863117.6	905502010.1	1119927395	1145943449	933147059.7	1099629664	1198361110	901382392.9	1171944559	1178942549	968494792.3	942861228.2	1021012067	534754328.6	974068768.3	1077247846	1029818690	1034823486	1066625401	1116051497	990947692.3	741165850.9	1023834783	1069729843	913606511.8	1026732998	1078694390	1007143848	1063117407	446379201.4	612169644.1
iso_Phe	912403196.4	867735996	857369490.3	881127275.6	886449431.9	597641232.5	322124517.2	816524266.6	856921487	228373278.2	837948901.1	839069745.9	721570576.5	803323541.8	773836230.9	721471375.7	764640963.5	752553347.3	746688756.4	846019472.8	877390215.1	746906706.8	838862528.1	904963276.8	731571432.8	868023019.9	878446098.1	789889607.2	777451989.4	844375152.6	433841595.8	825342193.8	878477564.3	835519256	845833504.1	873310022.3	887959488.8	832082222.4	636377175.3	839422201.9	854974229.8	759686754.7	849146174.7	870277887.9	827802232.4	855437136.3	357047507.9	530751366.3
PROLINE	3410531.092	295854562.7	309909891.3	2744108.622	285833098	266540306.7	1052693.31	288821146.7	244887009	923045.3002	296964476.7	253057089	61813128.88	308362517.5	336190346.3	57955665.5	313413127.4	337249869.7	6145188.916	2891206.218	5879467.792	2062844.336	2760217.406	3526512.367	1777508.161	2904234.622	5263151.745	2213964.552	173082005.2	265178318.4	195324119.2	179437111.1	258913842.3	278896722.3	175072384	244942452.7	253876862.2	181538201.8	178706293	287243086.2	247722758.7	194171195.4	258988744.9	248602817.3	194934441	255348043.3	148919253.4	174540814.9
iso_Pro	740509571	700260212.2	659851895.5	713443114.6	683846148.7	590129361.6	392867396	721099650.8	774001244.3	302633399.4	728083722.6	762976676.3	733764390.4	653940758.9	684487196.3	676093912.6	670594146.5	677015798.3	674368137.1	651381227.6	651366972.3	682616989.7	637940294	655036065	668995991.2	670739888.8	603207831.6	692265497.9	618644043.3	607869766.3	404923055.5	671163019.3	636619269.9	604581480.4	667656821.3	629657363.6	589226992.5	667243720.8	592491545.4	631913652.3	598158037.9	622073492.4	600939603.2	616748273.9	641879526.7	618565795.8	387626925.4	584272023.6
SERINE	51054219.78	25239091.15	25538131.54	45546850.03	25952412.05	19510380.4	18311097.69	20468675.9	19440657.82	13266231.67	22684211.72	20377176.48	41319975.25	28452469.52	26210434.12	43894204.68	28191778.54	26023188.44	33490484.99	40854131.27	43577794.53	32718542.71	39652961.29	44435131.9	33617531.66	42014344.46	42419482.25	35324913.47	23763316.51	23581775.71	14706509.35	26088482.38	25848419.91	24174701.21	29556420.17	27940957.24	27145901.33	25296740.16	19778394.22	23059690.41	25599593.81	23557120.09	23152319.37	26413863.27	25446006.86	26520976.89	14796480.61	15896171.15
iso_Ser	27701011.9	26406923.11	25890751.82	24624499.8	26914658.71	20155767.52	12026636.97	26256011	28181369.37	8552875.572	26681226.32	27065666.16	21472967.45	24462337.14	23444763.49	21955136.63	23218138.53	23261126.76	18993957.59	21627503.38	22994950.53	18223913.83	21058307.82	23281573.44	18608913.77	21745841.42	21675152.2	20021795.4	18724463.39	22199827.55	14388865.05	20934886.53	23457696.5	22627823.9	21943980.11	23957106.94	23358360.18	20242370.71	17491926.34	22589596.74	23123087.56	20150034.28	22613993.85	23748545.56	20610422.08	23157631.82	12762891.82	14941292.67
THREONINE	803058101.5	685725652.6	678601854.6	777966435	676925452.4	496477526.6	250376075.6	551081217.7	503691538.5	181418098.1	587722191.4	532231160.5	615263956.5	654063103.5	630277961.6	654504431.8	609581746.3	591083145.6	593234833.1	752158092.1	786613307.1	587462216.2	724398347.7	797425049.4	590463790	751378303.3	775060084.9	610621246.8	624038217.4	692503857	400899832.3	618218815.9	732984812.6	689662800.8	655802680.8	726217843.2	744277654.5	627943141.4	512547553.6	699532514.4	726113549.6	586019687.4	677515442.8	708482603.5	645632861.4	715585472	334711447	423970484.3
iso_Thr	217480876.7	207017764.8	198687770	214761215.1	201967470.1	147057296.1	84036074.99	195583869.5	212572759.6	58522534.8	197765384.6	204727553.6	171976162.7	191270098.6	187177144.7	167721821.3	179099980.5	175708444.1	172877765.1	204686953.5	214868668.9	171412403.6	198533243.7	221043189.7	172930393.2	202055967.1	206805163.1	184654108.1	180279757.1	202233500.1	115316608.2	187283405.2	211536103	200032836.9	194437931.8	210199441.9	216652966.1	190030416.2	158289299.1	207365272.7	207805341.5	176148640.6	200547949.7	202840782.7	192406867.4	203582969.8	97883072.99	133379686.7
TYROSINE	1137829619	990539765.1	1004105680	1126724335	1008665767	737789421	383512190.1	829106695.7	716643027.5	301881752.9	907087301.9	787311885	999515380.6	1000742399	953040832.5	1038580570	996688638.9	946545360.4	863904229.1	1068831481	1126992000	913140385.1	1057557422	1171325005	904472392.6	1140281670	1142239837	949382764.2	900052036.9	971602872.4	563595858.8	959674712.8	1026289916	997588286.4	1013360267	1019460015	1050980598	975686340	756036010.8	981801013	1016814356	890233232.6	964051273.1	1058579067	995103793.4	1024961787	477670034.6	641830037
iso_Tyr	722105137.7	684313838.9	675400483.3	710376503.6	682415571.8	498410981.7	289597175.1	669905214.1	687269147.5	216906502.8	689482981.4	686250558.2	629224792.5	671878603.8	642697535	626405650.1	655119248.2	640157370.5	584191366	662065572.9	709298943.6	601122539.3	662169641.4	726540136.6	604130301.8	696019025.6	697405416.3	637507536.3	604351506.6	648727450.5	369807168.8	665829790.9	684574908.2	658750583.7	679356012.8	684883094.4	684170217.7	666071856.5	531920761.7	653206729.6	663900890.6	607720465.2	651413361.8	693800645	669531028.5	669949083.1	312616620	454523673.9
VALINE	563224701.5	471423227.7	474426862	558946320.6	481440131	345710747.3	196772702.7	376027965.2	339107135.5	153683726.5	407516404.6	370098617.3	476987449.3	448167440.8	405853768.1	482945504.2	444224305.2	423930424.7	468503249	522456791	534526460.5	470856401.5	521798403.3	557903214.3	452801687.4	546118011.9	559579843.4	484814993.5	450822977.8	448341748.9	254693524.5	467104927	471635656.6	472080131	483561384.6	471554899.4	501691096.9	473556510.8	360972487.3	453613669	482026091.7	438746077.6	460748473.6	479765348	471370219.1	480559233.5	221957395.6	309949914.4
iso_Val	159396997.9	155773105.5	153905666.9	157784057.4	155818847.5	111335248.1	67160663.9	145883169.8	154012883.9	50330692.73	148788638.8	153250701.7	132235963	137332057.2	129080009.4	129676597.2	134977215.7	133357057.3	143149528.2	146598483.4	152603198.3	139390077.2	145133821.3	156344500.5	136720904.1	148507862.2	151986096.6	147173351	143929783.2	144088517.6	80147299.64	152525008.8	149787428	147343876	151603866.9	149161714.3	151821927.3	152597221	120507678.4	144705440	148801956.2	139880166.7	146303312.3	148163091.9	149745813	148799870.1	68594717.64	104778579.6
ISOLEUCINE	43358624.45	35481724.64	35513572.31	43543199.22	35834945.08	30666225.58	19662204.65	27907029.16	25596731.75	15327773.78	30548654.08	27947486.17	37028869.88	34069803.07	31487118.83	37689317.19	32920744.4	31758134.64	922011293.8	991612658.1	997226703.6	898539327.7	976864107	1043934542	836389306.3	1016445438	1011444759	930884798.8	865238024.2	836632295.2	437361772.7	872523625.1	861654614.3	856775996.1	909813912.8	854197698.2	902672182.2	890678957.9	671738306.1	830726221.7	859710869.7	801911471.7	831825212.1	864156472.1	864959414.1	881913583.8	372143961.8	562306776.6
iso_Ile	12338336.48	12434397.02	12362123	12374097.14	12233649.05	10545887.53	6679720.595	11547700.7	12389059.77	4955280.008	11735817.9	12350132.62	10526251.7	11465460.17	10977675.83	10332721.33	10920136.8	10825302.56	278349784.9	277841600.5	281702205.6	265861484.1	274354297.9	288537600.2	249320424.4	278079892.8	279335295.9	282191845.8	278161963.8	276434119.9	145671995.5	292217543	290712305.1	280871831	296402424.7	285215726.1	282400668.8	297378923.3	231413475.7	281401403.9	272842086.8	265715741.7	282970654.4	277446805.2	284297490.9	280476674.3	119438551.2	195794915.5
LEUCINE	35464862.18	28203244.05	28401891.38	34687373.55	29113701.4	26272364.83	17749135.05	22119111.14	20331865.95	14067109.62	23899040.04	22307222.36	30278999.03	26774953.94	24961529.25	30707878.24	26781828.57	25311193.66	812425389.2	889539468.2	912244303.8	808926775.5	888424914	937285142.4	760191612.3	929924875.5	934719844.7	838632157.7	741786534.4	716423835.6	369797393.4	766015725.8	768737610.3	732779851.7	792626985.6	773813308.7	790215289.4	778559356.1	575153860.8	727924698.3	739994246.6	691992929.5	734800351.5	759785525.6	761293907.3	762166101.7	320908962.2	479868467.7
iso_Leu	10280113.4	10650961.97	10390847.22	10079780.75	10418441.62	9610413.479	6220660.269	9788680.308	10429496.64	4658571.643	9843097.306	10255902.16	8667100.374	9526422.791	9023261.814	8517017.899	9175921.18	9014100.047	250233506.5	249892295.3	259839277.7	241367445.8	250253696.5	264058210.7	230349171.8	256228787.5	258085763.2	256074666.3	257799764.7	260572970.1	130525582.2	272942311	264493665.9	258279082.2	261690359.3	263038932.9	268374769.2	268670690.5	211851493.9	257141348.8	259321468.7	245140647.3	258084083.1	262657725.1	258109257.4	262424230.8	108429956.3	176874005.9
TRYPTOPHAN	195448895.7	161695282.2	163761934	191972129	169419936.9	117193685.9	57797809.46	132419143.8	117202276.9	42713050.45	144942824.3	124406472.6	164978050.5	163143200.5	153269845.6	173180004.7	161350193.6	151207738.2	138369641.3	168780964.9	180868966.9	146025839.8	170337175.3	191083879.8	141857826	184746572.3	185574977.2	150080474	148979995.9	148364069.2	78647280.24	157957861.8	161674800.1	154680900.4	165174508.8	160350859.9	171278970.2	160297618.7	116895313.5	148202809.9	156393081	140163266.7	153886912.8	165543283.6	162023768.6	163341901.4	68356419.56	96239478.73
MS_METABOLITE_DATA_END
#METABOLITES
METABOLITES_START
metabolite_name	Chemical Formula	RT	Mass (m/z)	Category
ALANINE / SARCOSINE	C3H7NO2	12.48	90.055	HMDB0000161 / HMDB0000271
iso_Ala	C3H7O2N	12.36	94.06183	standard1
ARGININE	C6H14N4O2	27.85	175.119	HMDB0000517
iso_Arg	C6H14N4O2	27.83	185.12675	standard2
ASPARAGINE	C4H8N2O3	13.15	133.0608	HMDB0000168
iso_Asn	C4H8N2O3	13.03	139.06784	standard3
ASPARTATE	C4H7NO4	12.55	132.0302	HMDB0000191
iso_Asp	C4H7NO4	12.85	137.04037	standard4
CYSTINE	C6H12N2O4S2	14.38	241.0317	HMDB0000192
iso_CysCys	C6H12N2O4S2	14.22	249.04469	standard5
GLUTAMINE	C5H10N2O3	12.98	147.0764	HMDB0000641
iso_Gln	C5H10N2O3	12.8	154.08685	standard6
GLUTAMATE	C5H9NO4	12.43	148.0604	HMDB0000148
iso_Glu	C5H9NO4	12.45	154.07376	standard7
GLYCINE	C2H5NO2	13.51	76.0393	HMDB0000123
iso_Gly	C2H5O2N	13.45	79.04284	standard8
LEUCINE	C6H13NO2	7.39	132.1019	HMDB0000687
iso_Leu	C6H13O2N	7.31	139.11873	standard11
ISOLEUCINE	C6H13NO2	8	132.1019	HMDB0000172
iso_Ile	C6H13O2N	8	139.11879	standard10
LYSINE	C6H14N2O2	26.05	147.1126	HMDB0000182
iso_Lys	C6H14N2O2	26.16	155.12656	standard12
METHIONINE	C5H11NO2S	8.29	150.0583	HMDB0000696
iso_Met	C5H11NO2S	8.25	156.07175	standard13
PHENYLALANINE	C9H11NO2	6.48	166.0863	HMDB0000159
iso_Phe	C9H11NO2	6.42	176.11308	standard14
PROLINE	C5H9NO2	9.83	116.0706	HMDB0000162
iso_Pro	C5H9NO2	9.82	122.08415	standard15
SERINE	C3H7NO3	13.71	104.0353	HMDB0000187
iso_Ser	C3H7O3N	13.6	108.04213	standard16
THREONINE	C4H9NO3	12.43	120.0655	HMDB0000167
iso_Thr	C4H9NO3	11.8	125.07566	standard17
TYROSINE	C9H11NO3	10.17	182.0812	HMDB0000158
iso_Tyr	C9H11NO3	10.05	192.10793	standard18
VALINE	C5H11NO2	9.53	118.0863	HMDB0000883
iso_Val	C5H11O2N	9.45	124.09978	standard19
TRYPTOPHAN	C11H12N2O2	8.33	205.0972	HMDB0013609
METABOLITES_END
#END