#METABOLOMICS WORKBENCH Sethjparker_20241101_174730 DATATRACK_ID:5334 STUDY_ID:ST003556 ANALYSIS_ID:AN005845 PROJECT_ID:PR002190
VERSION                          	1
CREATED_ON                       	11-07-2024
#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
PR:DOI                           	http://dx.doi.org/10.21228/M8JB9Q
#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 µL was added to 250 µL of
ST:STUDY_SUMMARY                 	ice cold 80% methanol containing 1 µL 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
ST:SUBMIT_DATE                   	2024-11-01
#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]Additional sample data
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_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           	-	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           	-	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           	-	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           	-	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           	-	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           	-	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_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           	-	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           	-	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
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           	-	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           	-	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           	-	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           	-	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           	-	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
#COLLECTION
CO:COLLECTION_SUMMARY            	5 µL 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 µL 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:INSTRUMENT_NAME               	Thermo Vanquish
CH:COLUMN_NAME                   	Merck SeQuant ZIC-pHILIC (150 x 2.1mm,5um)
CH:COLUMN_TEMPERATURE            	25°C
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 µL/min
CH:SOLVENT_A                     	100% Water; 10 mM Ammonium Carbonate, pH 9.0
CH:SOLVENT_B                     	100% Acetonitrile
CH:CHROMATOGRAPHY_TYPE           	HILIC
#ANALYSIS
AN:ANALYSIS_TYPE                 	MS
#MS
MS:INSTRUMENT_NAME               	Thermo Exploris 240
MS:INSTRUMENT_TYPE               	Orbitrap
MS:MS_TYPE                       	ESI
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 320°C, and vaporizer temperature 75°C. 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:ION_MODE                      	POSITIVE
#MS_METABOLITE_DATA
MS_METABOLITE_DATA:UNITS         	Ion counts
MS_METABOLITE_DATA_START
Samples	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	Tom_NMA_CM1	Tom_NMA_CM2	Tom_NMA_CM3	HEK293_TOM_0NMA_CM1	HEK293_TOM_0NMA_CM2	HEK293_TOM_0NMA_CM3	Tom_control_CM1	Tom_control_CM2	Tom_control_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	KO_NMA_CM1	KO_NMA_CM2	KO_NMA_CM3	HEK293_KO_0NMA_CM1	HEK293_KO_0NMA_CM2	HEK293_KO_0NMA_CM3	KO_control_CM1	KO_control_CM2	KO_control_CM3	DMEMt0_10NMA_CM1	DMEMt0_10NMA_CM2	DMEMt0_10NMA_CM3	DMEMt0_20NMA_CM1	DMEMt0_20NMA_CM2	DMEMt0_20NMA_CM3	CMt0_NMA_CM1	DMEMt0_5NMA_CM1	DMEMto_NMA_CM2	DMEMto_NMA_CM3	CMt0_control_CM1	DMEMt0_0NMA_CM1	DMEMt0_0NMA_CM2	DMEMt0_0NMA_CM3	DMEMto_control_CM2	DMEMto_control_CM3
Factors	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)	Sample source:Conditioned media | Genotype:CRISPR/Cas9 Control (sgTomato) | 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 Control (sgTomato) | 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: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 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 KO (sgALDH7A1) | 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:CRISPR/Cas9 KO (sgALDH7A1) | 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: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: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:None | Treatment:N-methyl-arginine (5 mM)	Sample source:Conditioned media | Genotype:None | Treatment:N-methyl-arginine (5 mM)	Sample source:Conditioned media | Genotype:None | 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:PBS	Sample source:Conditioned media | Genotype:None | Treatment:PBS	Sample source:Conditioned media | Genotype:None | Treatment:PBS	
ALANINE / SARCOSINE	67421442.5900	87757292.2200	81089304.1800	60184112.1700	73138513.8900	33983623.5300	60319401.3800	130752207.4000	110003426.9000	146034805.0000	75980512.3100	118632717.6000	120253847.6000	142568807.1000	117680159.8000	161157773.6000	67573953.0900	91800259.9000	101638036.1000	82280311.6800	78213951.2300	86966880.1900	80581254.9500	108772585.5000	98646306.0400	152867235.7000	86303434.5100	117544663.8000	78903076.7700	156145877.9000	100007832.0000	164047126.9000	2517430.0480	1865799.8290	2138095.6270	2009167.7790	2036702.6740	3424901.1660	1380953.8610	2081245.3260	331421.4837	65803807.5400	1912432.2060	3145078.8550	1922095.4250	4152468.0120	491806.2426	63523994.4400
ARGININE	677662186.1000	738287700.9000	801475915.0000	738071307.5000	760812577.6000	388049842.7000	474376331.6000	875249712.4000	837708787.6000	709931764.9000	556064706.1000	734699157.6000	822003965.2000	905123497.5000	754259471.4000	737174238.8000	718411391.0000	772983249.2000	768102098.9000	745697875.4000	762202968.7000	829849946.1000	738033712.8000	665159255.3000	747015049.8000	686533748.9000	714417709.4000	731152830.1000	444836837.9000	897202772.8000	684100962.6000	709756073.9000	733613172.8000	865936256.4000	970230916.3000	696322244.9000	865548472.5000	905853885.6000	1073343020.0000	776900427.2000	354383761.1000	851273799.4000	1176116454.0000	805660866.7000	970549644.9000	1049386077.0000	475774402.1000	886264416.5000
ASPARAGINE	4844871.1050	5552072.6140	4972792.8270	6097586.3040	6808428.9710	2962856.7610	3431311.1280	6173152.7760	9433385.0500	6069220.6410	4323331.7470	6051604.9240	5666553.1340	6637329.6380	7454180.4990	7079123.1420	4617134.9010	6797570.9550	6424680.9100	5472927.3100	8004001.0820	7703332.0680	4867362.5210	6294839.9030	5987441.4590	6644539.8710	4884736.6950	7678895.7850	4321409.2820	8412651.2750	5352816.5050	7357205.9670	87603.2779	87860.2505	91829.5246		69640.7634	148033.8898	154788.0386	76826.2535		1616791.6920	225081.4925	200700.5541	106560.0438	144628.5435	64810.2837	1742923.1080
ASPARTATE	461303.0064	672143.4083	672783.2901	379754.3479	921959.0303	304828.9931	665557.5816	189185.5287	672639.3240	895055.4657	380272.2430	606422.6623	519847.6186	753608.8014	482079.8712	654762.6191	1469759.5460	1002386.0990	1238163.6340	1078791.5680	1110331.7980	513958.3883	1216799.5300	421000.3300	245737.0336	712340.2617	1068525.6970	419195.5884	307563.5629	337780.9696	329323.3428	489362.9073	546021.1038	311762.9847	623405.4207	500320.5409	621984.3375	1042133.7320	407696.2881	713968.3674	106924.0573	5042934.8850	566591.3451	1085330.5690	787896.1154	963515.0973	151955.0791	4995623.0130
CYSTINE	136231261.3000	210028353.1000	227542585.6000	152583716.0000	213025576.3000	98333437.0000	85506722.6500	192033275.5000	156901125.1000	174286455.7000	115426827.4000	235333906.8000	253045790.7000	203100005.6000	145261880.9000	198948678.0000	128379773.3000	225626990.1000	199698274.4000	153527717.4000	223608168.4000	223924584.5000	136537297.4000	132248477.8000	127822869.2000	160582332.9000	143098829.8000	218631849.2000	116776661.7000	186942104.0000	119938016.7000	182298651.3000	137303989.2000	246803909.8000	284076978.0000	143973280.9000	259547735.7000	260770467.5000	232300790.5000	146706894.3000	45541866.5900	172426909.9000	238028321.1000	149611977.8000	278653918.0000	299110437.7000	63084118.3400	200004111.7000
GLUTAMATE	5898387.5050	8855849.9850	12284911.7000	7868532.1130	14121973.3800	3687732.8030	8184579.1140	4998996.8510	5812204.6570	21214322.0600	5008955.5730	6550111.0460	8226761.2600	10045281.2000	4824896.9770	12206628.5900	20931294.8800	16430106.1000	23424478.4700	12561962.8600	20139058.4900	10443342.3600	19597235.8000	6996321.3370	5892162.9540	17146967.2900	16180870.1600	7755405.5430	3953196.4020	7252539.0660	5151520.7500	8529758.2130	14518824.7300	17763670.0300	20660652.1700	13741328.0200	18167920.6700	24467168.8700	13885958.0200	15306167.8400	3929460.8220	107950164.2000	14820160.1200	25431847.4900	18469818.8900	20093968.7900	5367455.1130	96516053.1700
GLUTAMINE	2531090377.0000	2631305002.0000	2736328603.0000	2676157042.0000	2786217622.0000	1414296391.0000	1810274443.0000	2992909960.0000	2626775787.0000	2787713362.0000	2094980336.0000	2615092857.0000	2660810156.0000	2890756896.0000	2404715048.0000	2745208491.0000	2656320275.0000	2758596880.0000	2672903652.0000	2746946184.0000	2753638474.0000	2815732915.0000	2708803239.0000	2212736787.0000	2397159931.0000	2735184653.0000	2654079244.0000	2648580620.0000	1512119502.0000	2867774694.0000	2209244561.0000	2727126064.0000	2748224529.0000	3021028430.0000	3199548026.0000	2682917445.0000	3117873989.0000	3174715819.0000	3463250557.0000	2793408966.0000	1113632185.0000	3035711113.0000	3534371984.0000	2889923074.0000	3094072962.0000	3149348157.0000	1416827819.0000	2965641430.0000
GLYCINE	28187892.2500	28168275.2200	34386828.5300	32662960.2700	32759654.4600	10985613.8300	17512046.2100	26169642.3000	24105730.2000	25422753.3700	20882509.3200	30195992.6600	31009838.1500	28570736.6500	20755884.5500	25889067.7400	33134119.3800	31333967.2900	27919537.3800	29459483.9700	32934210.9900	33562841.8000	38111114.6400	27920872.8100	20316728.8400	25657293.4200	28117652.8200	30853879.1600	13889572.5400	25927507.1000	18750672.9200	25611053.1000	24246473.5400	27167469.7900	31367752.0900	23031044.8400	28748137.7200	32834358.1300	25563461.3700	37008463.9800	4569460.3150	42279963.9200	58211506.5400	58150202.5900	30402063.9200	29298532.5200	6172901.4810	21442443.7500
iso_Ala	166148003.4000	176863848.1000	183421163.5000	181974283.1000	183880542.0000	90668122.2200	125985959.8000	182412268.3000	172048612.2000	160850990.9000	141539941.5000	175980498.6000	186444740.1000	185692550.4000	171493975.5000	166016674.4000	179838985.3000	184853521.3000	179781484.2000	178269243.7000	187297976.8000	186116133.2000	182281318.1000	132160002.8000	179216948.9000	161715090.2000	168190975.4000	173197865.7000	102416669.3000	173426595.7000	181007096.4000	164488823.6000	173783202.9000	183632435.0000	196641532.3000	169535802.1000	188378231.7000	186200123.5000	195479062.8000	186107430.5000	60764978.7100	161551795.8000	201083782.0000	174553755.1000	191643460.4000	195934588.5000	83270305.7100	161733188.3000
iso_Arg	529058477.8000	602275311.1000	639943510.6000	543360035.8000	571653069.3000	297579501.5000	394836876.1000	627761046.8000	688339844.5000	532213095.2000	485827745.9000	668859455.8000	726252688.0000	726728626.6000	733063610.4000	616323376.4000	565698696.3000	618733503.6000	630454464.7000	545113647.1000	582949201.5000	623893663.3000	578437949.5000	488952629.3000	705331375.8000	541590387.4000	583037110.8000	655792400.8000	412669172.4000	708020630.6000	780707587.8000	603894002.6000	509411939.5000	584439448.0000	658044598.6000	485122379.4000	564579059.3000	600022262.2000	631743968.1000	546896124.7000	247185976.2000	526664769.7000	697716766.7000	573002593.0000	653095368.7000	720552009.2000	346989291.7000	566988348.2000
iso_Asn	72890162.4100	83183343.5700	84666353.6300	79188331.0900	85324494.8400	41785419.0700	51949434.8300	80043255.3500	78304269.0400	76119353.7300	61422657.6300	82172708.5100	83593316.2100	80580192.0900	75322757.8500	77610096.3600	76691799.5100	86406548.4200	83517038.7900	80443612.1200	87810814.4100	88824944.3700	74786564.8800	60796019.7300	79259730.8000	75391791.2300	70378749.4000	83753660.0700	47313313.2800	77194669.4900	78344123.6800	75550118.3900	69939180.4200	79748184.0600	87237899.3900	72151413.3500	83689470.5700	86213210.8500	77949282.9900	72254065.4900	24459296.6300	70690821.2600	79966332.3900	67711501.7800	81722266.0700	84299961.3000	32941502.8600	70582249.1300
iso_Asp	108674054.5000	115781756.9000	111824758.4000	112944903.7000	116137521.7000	50484206.8200	82778722.1100	114278896.5000	111735569.7000	114763920.0000	98534196.9100	113142228.0000	111690885.0000	117151315.9000	109348415.9000	116191636.6000	119563325.4000	117054110.5000	110626354.9000	116263719.2000	116131004.8000	112735804.9000	115520034.6000	83421989.0000	118172856.7000	116912421.9000	116836208.2000	116355941.6000	62953106.1900	114383706.8000	117790408.8000	115306985.4000	111286947.2000	112878452.8000	115874034.0000	102618482.4000	115122832.5000	111768429.4000	117338717.7000	113878000.8000	35832589.0300	108656553.3000	116879079.9000	120817623.2000	113671395.5000	115647835.6000	49343021.3000	110985364.0000
iso_CysCys	85984606.5500	133392958.1000	140594701.3000	95949858.4800	134123909.8000	60980245.7500	56362816.5900	125458568.0000	118803679.3000	116848503.0000	75454468.8800	146980813.6000	155902206.7000	133653890.7000	117362957.0000	134965046.5000	83649656.5500	144454310.4000	128210047.3000	95642115.5000	146107211.8000	139592935.0000	88776665.6200	92589708.3700	117854025.8000	112548526.0000	89685097.4500	142371909.8000	76003513.3900	125852296.8000	126839729.0000	128270663.1000	82191423.6000	139921372.8000	157976770.4000	82870986.5200	141060241.7000	143550413.5000	133999441.2000	86991073.3200	32779468.8500	104248591.8000	140247303.9000	92361230.5600	159539137.0000	173327068.2000	47139081.3200	123390760.8000
iso_Gln	230508701.1000	245152635.1000	245385580.6000	233881867.9000	247484298.7000	126366105.7000	173489247.3000	266290502.0000	269101631.2000	245402464.1000	199557777.3000	246944572.6000	243933327.7000	264437118.6000	262858738.6000	246243934.9000	243135030.4000	253226394.5000	247884483.3000	236520103.0000	249569566.2000	249732350.1000	242404733.2000	199405542.4000	278953857.2000	246633412.5000	229976911.9000	251094580.1000	145529202.3000	257178308.9000	284327766.4000	248082527.7000	215443948.4000	228746569.7000	240201105.1000	212844878.5000	230136298.3000	234665444.4000	247628428.6000	223213665.0000	92557849.8300	223933889.0000	252526047.8000	228408354.4000	234252219.4000	238856942.4000	122078373.1000	224353684.8000
iso_Glu	259823269.4000	278918266.6000	290897487.8000	277891560.4000	284400849.1000	138941146.2000	200187083.6000	288363506.8000	272276349.4000	269160299.2000	233115890.7000	282129686.9000	297095779.6000	296328535.3000	269332107.9000	280830884.1000	277482679.5000	288831031.5000	283065108.3000	279210446.6000	283568464.7000	294738608.6000	280372357.6000	218063141.2000	280981179.8000	270808400.2000	272066339.1000	280254807.4000	167449826.9000	283958624.8000	284061969.0000	278160074.1000	259159462.9000	277316491.0000	302919871.1000	254125100.5000	284996639.2000	290240327.8000	301465993.1000	279869520.5000	84579239.2100	247243871.9000	302861191.6000	269514990.1000	281687472.3000	300332822.4000	118626257.4000	252214826.4000
iso_Gly	20511768.4700	21914564.7500	21249040.0500	23484632.4900	22333537.2900	8725347.3100	13679828.3400	19593763.2100	19782375.5000	16972716.2100	16644498.8600	22678284.0900	22884271.5700	21371314.9800	19750122.8600	18645925.5600	20844639.5100	23694977.4200	19714767.9900	21903577.2500	23442256.9000	23317370.3500	21725092.0100	13992362.4600	20019664.7000	17306295.0700	20626057.1300	22682369.3200	10489068.5100	19989469.8600	19875428.7400	18406435.1300	19566480.0000	21369594.5700	23581962.6000	19507034.0500	22569622.1800	21189744.6000	20701633.5200	20868043.6400	4823084.2380	15943820.5500	21562524.8700	20751276.3400	23535135.2900	22943355.4800	6525293.4250	16254076.8200
iso_Ile	265715741.7000	282970654.4000	277446805.2000	284297490.9000	280476674.3000	119438551.2000	195794915.5000	12233649.0500	11735817.9000	10920136.8000	231413475.7000	281401403.9000	272842086.8000	12434397.0200	11547700.7000	11465460.1700	292217543.0000	290712305.1000	280871831.0000	296402424.7000	285215726.1000	282400668.8000	297378923.3000	10545887.5300	12350132.6200	10825302.5600	278161963.8000	276434119.9000	145671995.5000	12362123.0000	12389059.7700	10977675.8300	265861484.1000	274354297.9000	288537600.2000	249320424.4000	278079892.8000	279335295.9000	12374097.1400	282191845.8000	4955280.0080	10332721.3300	12338336.4800	278349784.9000	277841600.5000	281702205.6000	6679720.5950	10526251.7000
iso_Leu	245140647.3000	258084083.1000	262657725.1000	258109257.4000	262424230.8000	108429956.3000	176874005.9000	10418441.6200	9843097.3060	9175921.1800	211851493.9000	257141348.8000	259321468.7000	10650961.9700	9788680.3080	9526422.7910	272942311.0000	264493665.9000	258279082.2000	261690359.3000	263038932.9000	268374769.2000	268670690.5000	9610413.4790	10255902.1600	9014100.0470	257799764.7000	260572970.1000	130525582.2000	10390847.2200	10429496.6400	9023261.8140	241367445.8000	250253696.5000	264058210.7000	230349171.8000	256228787.5000	258085763.2000	10079780.7500	256074666.3000	4658571.6430	8517017.8990	10280113.4000	250233506.5000	249892295.3000	259839277.7000	6220660.2690	8667100.3740
ISOLEUCINE	801911471.7000	831825212.1000	864156472.1000	864959414.1000	881913583.8000	372143961.8000	562306776.6000	35834945.0800	30548654.0800	32920744.4000	671738306.1000	830726221.7000	859710869.7000	35481724.6400	27907029.1600	34069803.0700	872523625.1000	861654614.3000	856775996.1000	909813912.8000	854197698.2000	902672182.2000	890678957.9000	30666225.5800	27947486.1700	31758134.6400	865238024.2000	836632295.2000	437361772.7000	35513572.3100	25596731.7500	31487118.8300	898539327.7000	976864107.0000	1043934542.0000	836389306.3000	1016445438.0000	1011444759.0000	43543199.2200	930884798.8000	15327773.7800	37689317.1900	43358624.4500	922011293.8000	991612658.1000	997226703.6000	19662204.6500	37028869.8800
iso_Lys	62688142.8200	69486240.7900	71229339.6400	62763602.0200	64502547.9300	30386315.4900	47355309.6400	67535914.8800	63290659.8700	59084705.6200	66343410.8500	96343952.2200	101703097.2000	96480396.7500	83227704.1700	79829672.4200	67870229.6600	71976630.0400	69766938.0900	64490167.2200	66987971.5700	67451883.1700	72500253.7900	51372506.2700	65458017.9100	57368506.4900	82061623.4000	88686507.3800	50818990.4200	92439401.5700	88398871.9700	77261796.4400	62585352.7800	68356589.5300	75090052.1000	58092882.2600	65477533.8800	67567025.6200	67741580.6800	70138100.4800	18332578.0400	56724702.2200	94595307.2700	82838641.0600	98082888.4600	106059363.8000	32881077.5400	73571139.0900
iso_Met	251332776.9000	275033224.4000	280412972.7000	269882141.3000	280034588.2000	120882904.1000	181696653.6000	304145578.9000	291824995.0000	267833390.4000	213593605.3000	274559965.0000	276733826.7000	304732056.9000	282476539.3000	279422219.3000	272120655.9000	286728137.6000	278072680.1000	274363949.0000	286125994.1000	284013002.8000	275206824.7000	213705077.5000	296732158.6000	267940611.5000	256013628.8000	273139413.1000	144276765.4000	298311709.9000	299240525.0000	270898569.5000	243123968.4000	268492291.4000	283465859.0000	237387671.1000	272379319.6000	275061796.0000	298218308.9000	258207133.2000	83250279.4900	248829555.4000	306231555.5000	243616576.7000	269867711.0000	278032642.9000	115877659.3000	252265266.5000
iso_Phe	759686754.7000	849146174.7000	870277887.9000	827802232.4000	855437136.3000	357047507.9000	530751366.3000	886449431.9000	837948901.1000	764640963.5000	636377175.3000	839422201.9000	854974229.8000	867735996.0000	816524266.6000	803323541.8000	825342193.8000	878477564.3000	835519256.0000	845833504.1000	873310022.3000	887959488.8000	832082222.4000	597641232.5000	839069745.9000	752553347.3000	777451989.4000	844375152.6000	433841595.8000	857369490.3000	856921487.0000	773836230.9000	746906706.8000	838862528.1000	904963276.8000	731571432.8000	868023019.9000	878446098.1000	881127275.6000	789889607.2000	228373278.2000	721471375.7000	912403196.4000	746688756.4000	846019472.8000	877390215.1000	322124517.2000	721570576.5000
iso_Pro	622073492.4000	600939603.2000	616748273.9000	641879526.7000	618565795.8000	387626925.4000	584272023.6000	683846148.7000	728083722.6000	670594146.5000	592491545.4000	631913652.3000	598158037.9000	700260212.2000	721099650.8000	653940758.9000	671163019.3000	636619269.9000	604581480.4000	667656821.3000	629657363.6000	589226992.5000	667243720.8000	590129361.6000	762976676.3000	677015798.3000	618644043.3000	607869766.3000	404923055.5000	659851895.5000	774001244.3000	684487196.3000	682616989.7000	637940294.0000	655036065.0000	668995991.2000	670739888.8000	603207831.6000	713443114.6000	692265497.9000	302633399.4000	676093912.6000	740509571.0000	674368137.1000	651381227.6000	651366972.3000	392867396.0000	733764390.4000
iso_Ser	20150034.2800	22613993.8500	23748545.5600	20610422.0800	23157631.8200	12762891.8200	14941292.6700	26914658.7100	26681226.3200	23218138.5300	17491926.3400	22589596.7400	23123087.5600	26406923.1100	26256011.0000	24462337.1400	20934886.5300	23457696.5000	22627823.9000	21943980.1100	23957106.9400	23358360.1800	20242370.7100	20155767.5200	27065666.1600	23261126.7600	18724463.3900	22199827.5500	14388865.0500	25890751.8200	28181369.3700	23444763.4900	18223913.8300	21058307.8200	23281573.4400	18608913.7700	21745841.4200	21675152.2000	24624499.8000	20021795.4000	8552875.5720	21955136.6300	27701011.9000	18993957.5900	21627503.3800	22994950.5300	12026636.9700	21472967.4500
iso_Thr	176148640.6000	200547949.7000	202840782.7000	192406867.4000	203582969.8000	97883072.9900	133379686.7000	201967470.1000	197765384.6000	179099980.5000	158289299.1000	207365272.7000	207805341.5000	207017764.8000	195583869.5000	191270098.6000	187283405.2000	211536103.0000	200032836.9000	194437931.8000	210199441.9000	216652966.1000	190030416.2000	147057296.1000	204727553.6000	175708444.1000	180279757.1000	202233500.1000	115316608.2000	198687770.0000	212572759.6000	187177144.7000	171412403.6000	198533243.7000	221043189.7000	172930393.2000	202055967.1000	206805163.1000	214761215.1000	184654108.1000	58522534.8000	167721821.3000	217480876.7000	172877765.1000	204686953.5000	214868668.9000	84036074.9900	171976162.7000
iso_Tyr	607720465.2000	651413361.8000	693800645.0000	669531028.5000	669949083.1000	312616620.0000	454523673.9000	682415571.8000	689482981.4000	655119248.2000	531920761.7000	653206729.6000	663900890.6000	684313838.9000	669905214.1000	671878603.8000	665829790.9000	684574908.2000	658750583.7000	679356012.8000	684883094.4000	684170217.7000	666071856.5000	498410981.7000	686250558.2000	640157370.5000	604351506.6000	648727450.5000	369807168.8000	675400483.3000	687269147.5000	642697535.0000	601122539.3000	662169641.4000	726540136.6000	604130301.8000	696019025.6000	697405416.3000	710376503.6000	637507536.3000	216906502.8000	626405650.1000	722105137.7000	584191366.0000	662065572.9000	709298943.6000	289597175.1000	629224792.5000
iso_Val	139880166.7000	146303312.3000	148163091.9000	149745813.0000	148799870.1000	68594717.6400	104778579.6000	155818847.5000	148788638.8000	134977215.7000	120507678.4000	144705440.0000	148801956.2000	155773105.5000	145883169.8000	137332057.2000	152525008.8000	149787428.0000	147343876.0000	151603866.9000	149161714.3000	151821927.3000	152597221.0000	111335248.1000	153250701.7000	133357057.3000	143929783.2000	144088517.6000	80147299.6400	153905666.9000	154012883.9000	129080009.4000	139390077.2000	145133821.3000	156344500.5000	136720904.1000	148507862.2000	151986096.6000	157784057.4000	147173351.0000	50330692.7300	129676597.2000	159396997.9000	143149528.2000	146598483.4000	152603198.3000	67160663.9000	132235963.0000
LEUCINE	691992929.5000	734800351.5000	759785525.6000	761293907.3000	762166101.7000	320908962.2000	479868467.7000	29113701.4000	23899040.0400	26781828.5700	575153860.8000	727924698.3000	739994246.6000	28203244.0500	22119111.1400	26774953.9400	766015725.8000	768737610.3000	732779851.7000	792626985.6000	773813308.7000	790215289.4000	778559356.1000	26272364.8300	22307222.3600	25311193.6600	741786534.4000	716423835.6000	369797393.4000	28401891.3800	20331865.9500	24961529.2500	808926775.5000	888424914.0000	937285142.4000	760191612.3000	929924875.5000	934719844.7000	34687373.5500	838632157.7000	14067109.6200	30707878.2400	35464862.1800	812425389.2000	889539468.2000	912244303.8000	17749135.0500	30278999.0300
LYSINE	194361742.9000	217412585.3000	230438467.4000	199145266.5000	209571113.2000	97663869.8600	141443299.5000	229808006.5000	196568740.5000	216125361.8000	200352934.3000	304599037.8000	330956989.5000	320484067.2000	239609611.7000	289842131.4000	208295489.2000	226285682.4000	223117143.2000	206013698.3000	211270867.0000	219311691.6000	223461039.3000	173492527.9000	172685900.1000	203182029.3000	261469063.0000	276815199.6000	161535197.1000	314071496.0000	212455424.6000	275526003.9000	203343086.0000	234721793.0000	257578665.0000	186425515.8000	229510971.1000	237417357.1000	254303987.4000	222933808.5000	59589169.7000	227863733.3000	356763811.0000	268907005.4000	342217164.3000	366465713.9000	104111541.5000	291728267.8000
METHIONINE	187065861.4000	203754614.2000	213758632.6000	205721183.6000	216902594.9000	93561129.7100	130204120.6000	224454869.6000	191828356.7000	204856324.9000	153680978.5000	205510589.9000	210465958.1000	220112252.4000	172135374.6000	208086850.4000	201595823.0000	216312884.0000	207251558.0000	211622728.5000	217345016.5000	220955919.7000	207497207.9000	158275889.4000	171056517.9000	196023740.4000	195625037.6000	202393262.3000	108209652.7000	222102927.7000	156939312.5000	197241033.4000	198419997.5000	231430892.0000	245717707.4000	190852635.8000	240593266.5000	242758024.8000	259150980.2000	207574123.3000	62806574.8400	223086245.4000	265344037.5000	193912569.7000	234149972.4000	238798668.3000	84767998.0000	216360114.6000
PHENYLALANINE	913606511.8000	1026732998.0000	1078694390.0000	1007143848.0000	1063117407.0000	446379201.4000	612169644.1000	1068780480.0000	903243577.5000	962293677.4000	741165850.9000	1023834783.0000	1069729843.0000	1028645284.0000	819894972.7000	988523164.1000	974068768.3000	1077247846.0000	1029818690.0000	1034823486.0000	1066625401.0000	1116051497.0000	990947692.3000	730918342.5000	795948579.7000	910863117.6000	942861228.2000	1021012067.0000	534754328.6000	1043818594.0000	735768470.3000	945335838.6000	933147059.7000	1099629664.0000	1198361110.0000	901382392.9000	1171944559.0000	1178942549.0000	1163189446.0000	968494792.3000	261888276.0000	987333426.2000	1198550110.0000	905502010.1000	1119927395.0000	1145943449.0000	353441457.8000	949154929.3000
PROLINE	194171195.4000	258988744.9000	248602817.3000	194934441.0000	255348043.3000	148919253.4000	174540814.9000	285833098.0000	296964476.7000	313413127.4000	178706293.0000	287243086.2000	247722758.7000	295854562.7000	288821146.7000	308362517.5000	179437111.1000	258913842.3000	278896722.3000	175072384.0000	244942452.7000	253876862.2000	181538201.8000	266540306.7000	253057089.0000	337249869.7000	173082005.2000	265178318.4000	195324119.2000	309909891.3000	244887009.0000	336190346.3000	2062844.3360	2760217.4060	3526512.3670	1777508.1610	2904234.6220	5263151.7450	2744108.6220	2213964.5520	923045.3002	57955665.5000	3410531.0920	6145188.9160	2891206.2180	5879467.7920	1052693.3100	61813128.8800
SERINE	23557120.0900	23152319.3700	26413863.2700	25446006.8600	26520976.8900	14796480.6100	15896171.1500	25952412.0500	22684211.7200	28191778.5400	19778394.2200	23059690.4100	25599593.8100	25239091.1500	20468675.9000	28452469.5200	26088482.3800	25848419.9100	24174701.2100	29556420.1700	27940957.2400	27145901.3300	25296740.1600	19510380.4000	20377176.4800	26023188.4400	23763316.5100	23581775.7100	14706509.3500	25538131.5400	19440657.8200	26210434.1200	32718542.7100	39652961.2900	44435131.9000	33617531.6600	42014344.4600	42419482.2500	45546850.0300	35324913.4700	13266231.6700	43894204.6800	51054219.7800	33490484.9900	40854131.2700	43577794.5300	18311097.6900	41319975.2500
THREONINE	586019687.4000	677515442.8000	708482603.5000	645632861.4000	715585472.0000	334711447.0000	423970484.3000	676925452.4000	587722191.4000	609581746.3000	512547553.6000	699532514.4000	726113549.6000	685725652.6000	551081217.7000	654063103.5000	618218815.9000	732984812.6000	689662800.8000	655802680.8000	726217843.2000	744277654.5000	627943141.4000	496477526.6000	532231160.5000	591083145.6000	624038217.4000	692503857.0000	400899832.3000	678601854.6000	503691538.5000	630277961.6000	587462216.2000	724398347.7000	797425049.4000	590463790.0000	751378303.3000	775060084.9000	777966435.0000	610621246.8000	181418098.1000	654504431.8000	803058101.5000	593234833.1000	752158092.1000	786613307.1000	250376075.6000	615263956.5000
TRYPTOPHAN	140163266.7000	153886912.8000	165543283.6000	162023768.6000	163341901.4000	68356419.5600	96239478.7300	169419936.9000	144942824.3000	161350193.6000	116895313.5000	148202809.9000	156393081.0000	161695282.2000	132419143.8000	163143200.5000	157957861.8000	161674800.1000	154680900.4000	165174508.8000	160350859.9000	171278970.2000	160297618.7000	117193685.9000	124406472.6000	151207738.2000	148979995.9000	148364069.2000	78647280.2400	163761934.0000	117202276.9000	153269845.6000	146025839.8000	170337175.3000	191083879.8000	141857826.0000	184746572.3000	185574977.2000	191972129.0000	150080474.0000	42713050.4500	173180004.7000	195448895.7000	138369641.3000	168780964.9000	180868966.9000	57797809.4600	164978050.5000
TYROSINE	890233232.6000	964051273.1000	1058579067.0000	995103793.4000	1024961787.0000	477670034.6000	641830037.0000	1008665767.0000	907087301.9000	996688638.9000	756036010.8000	981801013.0000	1016814356.0000	990539765.1000	829106695.7000	1000742399.0000	959674712.8000	1026289916.0000	997588286.4000	1013360267.0000	1019460015.0000	1050980598.0000	975686340.0000	737789421.0000	787311885.0000	946545360.4000	900052036.9000	971602872.4000	563595858.8000	1004105680.0000	716643027.5000	953040832.5000	913140385.1000	1057557422.0000	1171325005.0000	904472392.6000	1140281670.0000	1142239837.0000	1126724335.0000	949382764.2000	301881752.9000	1038580570.0000	1137829619.0000	863904229.1000	1068831481.0000	1126992000.0000	383512190.1000	999515380.6000
VALINE	438746077.6000	460748473.6000	479765348.0000	471370219.1000	480559233.5000	221957395.6000	309949914.4000	481440131.0000	407516404.6000	444224305.2000	360972487.3000	453613669.0000	482026091.7000	471423227.7000	376027965.2000	448167440.8000	467104927.0000	471635656.6000	472080131.0000	483561384.6000	471554899.4000	501691096.9000	473556510.8000	345710747.3000	370098617.3000	423930424.7000	450822977.8000	448341748.9000	254693524.5000	474426862.0000	339107135.5000	405853768.1000	470856401.5000	521798403.3000	557903214.3000	452801687.4000	546118011.9000	559579843.4000	558946320.6000	484814993.5000	153683726.5000	482945504.2000	563224701.5000	468503249.0000	522456791.0000	534526460.5000	196772702.7000	476987449.3000
MS_METABOLITE_DATA_END
#METABOLITES
METABOLITES_START
metabolite_name	pubchem_id	inchi_key	kegg_id	other_id	other_id_type	ri	ri_type	moverz_quant	
ALANINE / SARCOSINE						12.48		90.055	
ARGININE						27.85		175.119	
ASPARAGINE						13.15		133.0608	
ASPARTATE						12.55		132.0302	
CYSTINE						14.38		241.0317	
GLUTAMATE						12.43		148.0604	
GLUTAMINE						12.98		147.0764	
GLYCINE						13.51		76.0393	
iso_Ala						12.36		94.06183	
iso_Arg						27.83		185.12675	
iso_Asn						13.03		139.06784	
iso_Asp						12.85		137.04037	
iso_CysCys						14.22		249.04469	
iso_Gln						12.8		154.08685	
iso_Glu						12.45		154.07376	
iso_Gly						13.45		79.04284	
iso_Ile						8		139.11879	
iso_Leu						7.31		139.11873	
ISOLEUCINE						8		132.1019	
iso_Lys						26.16		155.12656	
iso_Met						8.25		156.07175	
iso_Phe						6.42		176.11308	
iso_Pro						9.82		122.08415	
iso_Ser						13.6		108.04213	
iso_Thr						11.8		125.07566	
iso_Tyr						10.05		192.10793	
iso_Val						9.45		124.09978	
LEUCINE						7.39		132.1019	
LYSINE						26.05		147.1126	
METHIONINE						8.29		150.0583	
PHENYLALANINE						6.48		166.0863	
PROLINE						9.83		116.0706	
SERINE						13.71		104.0353	
THREONINE						12.43		120.0655	
TRYPTOPHAN						8.33		205.0972	
TYROSINE						10.17		182.0812	
VALINE						9.53		118.0863	
METABOLITES_END
#END