#METABOLOMICS WORKBENCH tocunningham_20241219_113248 DATATRACK_ID:5467 STUDY_ID:ST003634 ANALYSIS_ID:AN005967 PROJECT_ID:PR002247
VERSION             	1
CREATED_ON             	December 23, 2024, 5:58 pm
#PROJECT
PR:PROJECT_TITLE                 	Untargeted NMR-metabolomic study in NIH3T3 (mouse embryonic fibroblasts)
PR:PROJECT_SUMMARY               	Phosphoribosyl pyrophosphate synthetase (PRPS) is a highly conserved enzyme that
PR:PROJECT_SUMMARY               	conducts the chokepoint reaction of nucleotide biosynthesis by converting
PR:PROJECT_SUMMARY               	ribose-5-phosphate (R-5-P) to phosphoribosyl pyrophosphate (PRPP). Due to the
PR:PROJECT_SUMMARY               	presence of multiple isoforms and related PRPS-associated proteins in
PR:PROJECT_SUMMARY               	opisthokont species, the precise nature of the PRPS enzyme in cells and tissues
PR:PROJECT_SUMMARY               	remains uncertain. Using proteomics approaches and biochemistry in mammalian
PR:PROJECT_SUMMARY               	cells, we demonstrate that these individual components assemble together to form
PR:PROJECT_SUMMARY               	a heterogeneous megadalton complex comprising of PRPS1 (P1), PRPS2 (P2), and two
PR:PROJECT_SUMMARY               	PRPS associated proteins – PRPSAP1 (AP1) and PRPSAP2 (AP2). Employing
PR:PROJECT_SUMMARY               	CRISPR-Cas9 knockout (KO) strategy in NIH3T3 cells (mouse embryonic fibroblasts)
PR:PROJECT_SUMMARY               	to generate all viable genetic knockout combinations, we observed that
PR:PROJECT_SUMMARY               	PRPS1-only expressing cells (P2/AP1/AP2 triple KO) has severe cellular
PR:PROJECT_SUMMARY               	proliferation defects as compared to other knockout lines. To pinpoint the
PR:PROJECT_SUMMARY               	metabolic basis for decreased proliferative capacity of PRPS1-only expressing
PR:PROJECT_SUMMARY               	cells (P2/AP1/AP2 KO), we used NMR based 13C isotopic tracing to compare the
PR:PROJECT_SUMMARY               	metabolic state of wildtype and P2/AP1/AP2 KO NIH3T3 cells. The results
PR:PROJECT_SUMMARY               	demonstrate that P2/AP1/AP2 KO cells exhibit reduced pentose phosphate pathway
PR:PROJECT_SUMMARY               	flux and nucleotide production, alongside alterations in glycolysis, the
PR:PROJECT_SUMMARY               	creatine phosphate pathway, and choline metabolism. These changes reflect
PR:PROJECT_SUMMARY               	decreased PRPP synthesis rates, likely due to suboptimal PRPS1 homotypic
PR:PROJECT_SUMMARY               	configurations. Collectively, this study revealed that PRPS1 as a standalone
PR:PROJECT_SUMMARY               	enzyme in cells is insufficient to sustain the metabolic flux necessary to keep
PR:PROJECT_SUMMARY               	up with the demands of proliferation.
PR:INSTITUTE                     	University of Cincinnati College of Medicine
PR:DEPARTMENT                    	Department of Cancer Biology
PR:LABORATORY                    	Cunningham Lab
PR:LAST_NAME                     	Cunningham
PR:FIRST_NAME                    	John
PR:ADDRESS                       	3125 Eden Avenue, Vontz Center for Molecular Studies Rm 3306, Cincinnati, OH
PR:ADDRESS                       	45219, USA
PR:EMAIL                         	cunnijn@ucmail.uc.edu
PR:PHONE                         	513-558-1157
PR:CONTRIBUTORS                  	Bibek R. Karki, Sara Vicente-Muñoz, Lindsey E. Romick and J. Tom Cunningham
#STUDY
ST:STUDY_TITLE                   	NMR metabolomics confirms PRPS homooligomerization in PRPS2/PRPSAP1/PRPSAP2
ST:STUDY_TITLE                   	triple knockout mouse embryonic fibroblasts decreases flux through the nucletide
ST:STUDY_TITLE                   	biosynthesis pathways
ST:STUDY_SUMMARY                 	Phosphoribosyl pyrophosphate synthetase (PRPS) is a highly conserved enzyme that
ST:STUDY_SUMMARY                 	conducts the chokepoint reaction of nucleotide biosynthesis by converting
ST:STUDY_SUMMARY                 	ribose-5-phosphate (R-5-P) to phosphoribosyl pyrophosphate (PRPP). Due to the
ST:STUDY_SUMMARY                 	presence of multiple isoforms and related PRPS-associated proteins in
ST:STUDY_SUMMARY                 	opisthokont species, the precise nature of the PRPS enzyme in cells and tissues
ST:STUDY_SUMMARY                 	remains uncertain. Using proteomics approaches and biochemistry in mammalian
ST:STUDY_SUMMARY                 	cells, we demonstrate that these individual components assemble together to form
ST:STUDY_SUMMARY                 	a heterogeneous megadalton complex comprising of PRPS1 (P1), PRPS2 (P2), and two
ST:STUDY_SUMMARY                 	PRPS associated proteins – PRPSAP1 (AP1) and PRPSAP2 (AP2). Employing
ST:STUDY_SUMMARY                 	CRISPR-Cas9 knockout (KO) strategy in NIH3T3 cells (mouse embryonic fibroblasts)
ST:STUDY_SUMMARY                 	to generate all viable genetic knockout combinations, we observed that
ST:STUDY_SUMMARY                 	PRPS1-only expressing cells (P2/AP1/AP2 triple KO) has severe cellular
ST:STUDY_SUMMARY                 	proliferation defects as compared to other knockout lines. To pinpoint the
ST:STUDY_SUMMARY                 	metabolic basis for decreased proliferative capacity of PRPS1-only expressing
ST:STUDY_SUMMARY                 	cells (P2/AP1/AP2 KO), we used NMR based 13C isotopic tracing to compare the
ST:STUDY_SUMMARY                 	metabolic state of wildtype and P2/AP1/AP2 KO NIH3T3 cells. The results
ST:STUDY_SUMMARY                 	demonstrate that P2/AP1/AP2 KO cells exhibit reduced pentose phosphate pathway
ST:STUDY_SUMMARY                 	flux and nucleotide production, alongside alterations in glycolysis, the
ST:STUDY_SUMMARY                 	creatine phosphate pathway, and choline metabolism. These changes reflect
ST:STUDY_SUMMARY                 	decreased PRPP synthesis rates, likely due to suboptimal PRPS1 homotypic
ST:STUDY_SUMMARY                 	configurations. Collectively, this study revealed that PRPS1 as a standalone
ST:STUDY_SUMMARY                 	enzyme in cells is insufficient to sustain the metabolic flux necessary to keep
ST:STUDY_SUMMARY                 	up with the demands of proliferation.
ST:INSTITUTE                     	University of Cincinnati College of Medicine
ST:DEPARTMENT                    	Department of Cancer Biology
ST:LABORATORY                    	Cunningham Lab
ST:LAST_NAME                     	Cunningham
ST:FIRST_NAME                    	Tom
ST:ADDRESS                       	3125 Eden Avenue
ST:EMAIL                         	cunnijn@ucmail.uc.edu
ST:PHONE                         	513-558-1157
#SUBJECT
SU:SUBJECT_TYPE                  	Cultured cells
SU:SUBJECT_SPECIES               	Mus musculus
SU:TAXONOMY_ID                   	10090
SU:GENDER                        	Male
SU:CELL_BIOSOURCE_OR_SUPPLIER    	NIH3T3 (CRL-1658) were obtained from American Type Culture Collection (ATCC,
SU:CELL_BIOSOURCE_OR_SUPPLIER    	Manassass, VA, USA)
SU:CELL_STRAIN_DETAILS           	NH3T3 (mouse embryonic fibroblasts)
SU:SUBJECT_COMMENTS              	ATCC-derived cultures are authenticated by morphological profiling, karyotyping,
SU:SUBJECT_COMMENTS              	and PCR-based methodologies (STR profiling).
#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           	-	Cunninghan_UC_PRPS_Cells_110222.101	Sample source:NIH3T3 | Genotype:Wildtype1_Gr1 | Treatment:U12C glucose | Treatment duration:30 mins	NMR Experiment=1H; RAW_FILE_NAME(Raw file name)=Cunninghan_UC_PRPS_Cells_110222.101.fid
SUBJECT_SAMPLE_FACTORS           	-	Cunninghan_UC_PRPS_Cells_110222.102	Sample source:NIH3T3 | Genotype:Wildtype2_Gr1 | Treatment:U12C glucose | Treatment duration:30 mins	NMR Experiment=1H; RAW_FILE_NAME(Raw file name)=Cunninghan_UC_PRPS_Cells_110222.102.fid
SUBJECT_SAMPLE_FACTORS           	-	Cunninghan_UC_PRPS_Cells_110222.103	Sample source:NIH3T3 | Genotype:Wildtype3_Gr1 | Treatment:U12C glucose | Treatment duration:30 mins	NMR Experiment=1H; RAW_FILE_NAME(Raw file name)=Cunninghan_UC_PRPS_Cells_110222.103.fid
SUBJECT_SAMPLE_FACTORS           	-	Cunninghan_UC_PRPS_Cells_110222.104	Sample source:NIH3T3 | Genotype:Wildtype1_Gr2 | Treatment:U13C glucose | Treatment duration:30 mins	NMR Experiment=1H; RAW_FILE_NAME(Raw file name)=Cunninghan_UC_PRPS_Cells_110222.104.fid
SUBJECT_SAMPLE_FACTORS           	-	Cunninghan_UC_PRPS_Cells_110222.105	Sample source:NIH3T3 | Genotype:Wildtype2_Gr2 | Treatment:U13C glucose | Treatment duration:30 mins	NMR Experiment=1H; RAW_FILE_NAME(Raw file name)=Cunninghan_UC_PRPS_Cells_110222.105.fid
SUBJECT_SAMPLE_FACTORS           	-	Cunninghan_UC_PRPS_Cells_110222.106	Sample source:NIH3T3 | Genotype:Wildtype3_Gr2 | Treatment:U13C glucose | Treatment duration:30 mins	NMR Experiment=1H; RAW_FILE_NAME(Raw file name)=Cunninghan_UC_PRPS_Cells_110222.106.fid
SUBJECT_SAMPLE_FACTORS           	-	Cunninghan_UC_PRPS_Cells_110222.107	Sample source:NIH3T3 | Genotype:Wildtype1-Gr3 | Treatment:U13C glucose | Treatment duration:5 hrs	NMR Experiment=1H; RAW_FILE_NAME(Raw file name)=Cunninghan_UC_PRPS_Cells_110222.107.fid
SUBJECT_SAMPLE_FACTORS           	-	Cunninghan_UC_PRPS_Cells_110222.108	Sample source:NIH3T3 | Genotype:Wildtype2-Gr3 | Treatment:U13C glucose | Treatment duration:5 hrs	NMR Experiment=1H; RAW_FILE_NAME(Raw file name)=Cunninghan_UC_PRPS_Cells_110222.108.fid
SUBJECT_SAMPLE_FACTORS           	-	Cunninghan_UC_PRPS_Cells_110222.109	Sample source:NIH3T3 | Genotype:Wildtype3-Gr3 | Treatment:U13C glucose | Treatment duration:5 hrs	NMR Experiment=1H; RAW_FILE_NAME(Raw file name)=Cunninghan_UC_PRPS_Cells_110222.109.fid
SUBJECT_SAMPLE_FACTORS           	-	Cunninghan_UC_PRPS_Cells_110222.110	Sample source:NIH3T3 | Genotype:P2/AP1/AP2 KO1_Gr4 | Treatment:U12C glucose | Treatment duration:30 mins	NMR Experiment=1H; RAW_FILE_NAME(Raw file name)=Cunninghan_UC_PRPS_Cells_110222.110.fid
SUBJECT_SAMPLE_FACTORS           	-	Cunninghan_UC_PRPS_Cells_110222.111	Sample source:NIH3T3 | Genotype:P2/AP1/AP2 KO2_Gr4 | Treatment:U12C glucose | Treatment duration:30 mins	NMR Experiment=1H; RAW_FILE_NAME(Raw file name)=Cunninghan_UC_PRPS_Cells_110222.111.fid
SUBJECT_SAMPLE_FACTORS           	-	Cunninghan_UC_PRPS_Cells_110222.112	Sample source:NIH3T3 | Genotype:P2/AP1/AP2 KO3_Gr4 | Treatment:U12C glucose | Treatment duration:30 mins	NMR Experiment=1H; RAW_FILE_NAME(Raw file name)=Cunninghan_UC_PRPS_Cells_110222.112.fid
SUBJECT_SAMPLE_FACTORS           	-	Cunninghan_UC_PRPS_Cells_110222.113	Sample source:NIH3T3 | Genotype:P2/AP1/AP2 KO1_Gr5 | Treatment:U13C glucose | Treatment duration:30 mins	NMR Experiment=1H; RAW_FILE_NAME(Raw file name)=Cunninghan_UC_PRPS_Cells_110222.113.fid
SUBJECT_SAMPLE_FACTORS           	-	Cunninghan_UC_PRPS_Cells_110222.114	Sample source:NIH3T3 | Genotype:P2/AP1/AP2 KO2_Gr5 | Treatment:U13C glucose | Treatment duration:30 mins	NMR Experiment=1H; RAW_FILE_NAME(Raw file name)=Cunninghan_UC_PRPS_Cells_110222.114.fid
SUBJECT_SAMPLE_FACTORS           	-	Cunninghan_UC_PRPS_Cells_110222.115	Sample source:NIH3T3 | Genotype:P2/AP1/AP2 KO3_Gr5 | Treatment:U13C glucose | Treatment duration:30 mins	NMR Experiment=1H; RAW_FILE_NAME(Raw file name)=Cunninghan_UC_PRPS_Cells_110222.115.fid
SUBJECT_SAMPLE_FACTORS           	-	Cunninghan_UC_PRPS_Cells_110222.116	Sample source:NIH3T3 | Genotype:P2/AP1/AP2 KO1_Gr6 | Treatment:U13C glucose | Treatment duration:5 hrs	NMR Experiment=1H; RAW_FILE_NAME(Raw file name)=Cunninghan_UC_PRPS_Cells_110222.116.fid
SUBJECT_SAMPLE_FACTORS           	-	Cunninghan_UC_PRPS_Cells_110222.117	Sample source:NIH3T3 | Genotype:P2/AP1/AP2 KO2_Gr6 | Treatment:U13C glucose | Treatment duration:5 hrs	NMR Experiment=1H; RAW_FILE_NAME(Raw file name)=Cunninghan_UC_PRPS_Cells_110222.117.fid
SUBJECT_SAMPLE_FACTORS           	-	Cunninghan_UC_PRPS_Cells_110222.118	Sample source:NIH3T3 | Genotype:P2/AP1/AP2 KO3_Gr6 | Treatment:U13C glucose | Treatment duration:5 hrs	NMR Experiment=1H; RAW_FILE_NAME(Raw file name)=Cunninghan_UC_PRPS_Cells_110222.118.fid
SUBJECT_SAMPLE_FACTORS           	-	Cunninghan_UC_PRPS_Cells_110222.201	Sample source:NIH3T3 | Genotype:Wildtype1_Gr1 | Treatment:U12C glucose | Treatment duration:30 mins	NMR Experiment=1D 1H-13C-HSQC; RAW_FILE_NAME(Raw file name)=Cunninghan_UC_PRPS_Cells_110222.201.fid
SUBJECT_SAMPLE_FACTORS           	-	Cunninghan_UC_PRPS_Cells_110222.202	Sample source:NIH3T3 | Genotype:Wildtype2_Gr1 | Treatment:U12C glucose | Treatment duration:30 mins	NMR Experiment=1D 1H-13C-HSQC; RAW_FILE_NAME(Raw file name)=Cunninghan_UC_PRPS_Cells_110222.202.fid
SUBJECT_SAMPLE_FACTORS           	-	Cunninghan_UC_PRPS_Cells_110222.203	Sample source:NIH3T3 | Genotype:Wildtype3_Gr1 | Treatment:U12C glucose | Treatment duration:30 mins	NMR Experiment=1D 1H-13C-HSQC; RAW_FILE_NAME(Raw file name)=Cunninghan_UC_PRPS_Cells_110222.203.fid
SUBJECT_SAMPLE_FACTORS           	-	Cunninghan_UC_PRPS_Cells_110222.204	Sample source:NIH3T3 | Genotype:Wildtype1_Gr2 | Treatment:U13C glucose | Treatment duration:30 mins	NMR Experiment=1D 1H-13C-HSQC; RAW_FILE_NAME(Raw file name)=Cunninghan_UC_PRPS_Cells_110222.204.fid
SUBJECT_SAMPLE_FACTORS           	-	Cunninghan_UC_PRPS_Cells_110222.205	Sample source:NIH3T3 | Genotype:Wildtype2_Gr2 | Treatment:U13C glucose | Treatment duration:30 mins	NMR Experiment=1D 1H-13C-HSQC; RAW_FILE_NAME(Raw file name)=Cunninghan_UC_PRPS_Cells_110222.205.fid
SUBJECT_SAMPLE_FACTORS           	-	Cunninghan_UC_PRPS_Cells_110222.206	Sample source:NIH3T3 | Genotype:Wildtype3_Gr2 | Treatment:U13C glucose | Treatment duration:30 mins	NMR Experiment=1D 1H-13C-HSQC; RAW_FILE_NAME(Raw file name)=Cunninghan_UC_PRPS_Cells_110222.206.fid
SUBJECT_SAMPLE_FACTORS           	-	Cunninghan_UC_PRPS_Cells_110222.207	Sample source:NIH3T3 | Genotype:Wildtype1-Gr3 | Treatment:U13C glucose | Treatment duration:5 hrs	NMR Experiment=1D 1H-13C-HSQC; RAW_FILE_NAME(Raw file name)=Cunninghan_UC_PRPS_Cells_110222.207.fid
SUBJECT_SAMPLE_FACTORS           	-	Cunninghan_UC_PRPS_Cells_110222.208	Sample source:NIH3T3 | Genotype:Wildtype2-Gr3 | Treatment:U13C glucose | Treatment duration:5 hrs	NMR Experiment=1D 1H-13C-HSQC; RAW_FILE_NAME(Raw file name)=Cunninghan_UC_PRPS_Cells_110222.208.fid
SUBJECT_SAMPLE_FACTORS           	-	Cunninghan_UC_PRPS_Cells_110222.209	Sample source:NIH3T3 | Genotype:Wildtype3-Gr3 | Treatment:U13C glucose | Treatment duration:5 hrs	NMR Experiment=1D 1H-13C-HSQC; RAW_FILE_NAME(Raw file name)=Cunninghan_UC_PRPS_Cells_110222.209.fid
SUBJECT_SAMPLE_FACTORS           	-	Cunninghan_UC_PRPS_Cells_110222.210	Sample source:NIH3T3 | Genotype:P2/AP1/AP2 KO1_Gr4 | Treatment:U12C glucose | Treatment duration:30 mins	NMR Experiment=1D 1H-13C-HSQC; RAW_FILE_NAME(Raw file name)=Cunninghan_UC_PRPS_Cells_110222.210.fid
SUBJECT_SAMPLE_FACTORS           	-	Cunninghan_UC_PRPS_Cells_110222.211	Sample source:NIH3T3 | Genotype:P2/AP1/AP2 KO2_Gr4 | Treatment:U12C glucose | Treatment duration:30 mins	NMR Experiment=1D 1H-13C-HSQC; RAW_FILE_NAME(Raw file name)=Cunninghan_UC_PRPS_Cells_110222.211.fid
SUBJECT_SAMPLE_FACTORS           	-	Cunninghan_UC_PRPS_Cells_110222.212	Sample source:NIH3T3 | Genotype:P2/AP1/AP2 KO3_Gr4 | Treatment:U12C glucose | Treatment duration:30 mins	NMR Experiment=1D 1H-13C-HSQC; RAW_FILE_NAME(Raw file name)=Cunninghan_UC_PRPS_Cells_110222.212.fid
SUBJECT_SAMPLE_FACTORS           	-	Cunninghan_UC_PRPS_Cells_110222.213	Sample source:NIH3T3 | Genotype:P2/AP1/AP2 KO1_Gr5 | Treatment:U13C glucose | Treatment duration:30 mins	NMR Experiment=1D 1H-13C-HSQC; RAW_FILE_NAME(Raw file name)=Cunninghan_UC_PRPS_Cells_110222.213.fid
SUBJECT_SAMPLE_FACTORS           	-	Cunninghan_UC_PRPS_Cells_110222.214	Sample source:NIH3T3 | Genotype:P2/AP1/AP2 KO2_Gr5 | Treatment:U13C glucose | Treatment duration:30 mins	NMR Experiment=1D 1H-13C-HSQC; RAW_FILE_NAME(Raw file name)=Cunninghan_UC_PRPS_Cells_110222.214.fid
SUBJECT_SAMPLE_FACTORS           	-	Cunninghan_UC_PRPS_Cells_110222.215	Sample source:NIH3T3 | Genotype:P2/AP1/AP2 KO3_Gr5 | Treatment:U13C glucose | Treatment duration:30 mins	NMR Experiment=1D 1H-13C-HSQC; RAW_FILE_NAME(Raw file name)=Cunninghan_UC_PRPS_Cells_110222.215.fid
SUBJECT_SAMPLE_FACTORS           	-	Cunninghan_UC_PRPS_Cells_110222.216	Sample source:NIH3T3 | Genotype:P2/AP1/AP2 KO1_Gr6 | Treatment:U13C glucose | Treatment duration:5 hrs	NMR Experiment=1D 1H-13C-HSQC; RAW_FILE_NAME(Raw file name)=Cunninghan_UC_PRPS_Cells_110222.216.fid
SUBJECT_SAMPLE_FACTORS           	-	Cunninghan_UC_PRPS_Cells_110222.217	Sample source:NIH3T3 | Genotype:P2/AP1/AP2 KO2_Gr6 | Treatment:U13C glucose | Treatment duration:5 hrs	NMR Experiment=1D 1H-13C-HSQC; RAW_FILE_NAME(Raw file name)=Cunninghan_UC_PRPS_Cells_110222.217.fid
SUBJECT_SAMPLE_FACTORS           	-	Cunninghan_UC_PRPS_Cells_110222.218	Sample source:NIH3T3 | Genotype:P2/AP1/AP2 KO3_Gr6 | Treatment:U13C glucose | Treatment duration:5 hrs	NMR Experiment=1D 1H-13C-HSQC; RAW_FILE_NAME(Raw file name)=Cunninghan_UC_PRPS_Cells_110222.218.fid
#COLLECTION
CO:COLLECTION_SUMMARY            	NIH3T3 fibroblasts were seeded in 15 cm plates. Cells were maintained in
CO:COLLECTION_SUMMARY            	Dulbecco’s modified Eagle medium (DMEM) containing 10% (v/v) fetal bovine
CO:COLLECTION_SUMMARY            	serum (FBS) and 1x penicillin/streptomycin. The media were replaced with
CO:COLLECTION_SUMMARY            	glucose-free media containing 10 mM of 12C6-glucose (unlabeled; ThermoFisher
CO:COLLECTION_SUMMARY            	#410955000, Lot#B0144488) or 13C6-glucose (Cambridge Isotope Laboratories;
CO:COLLECTION_SUMMARY            	CLM-1396-50, Lot#PB-28582), supplemented with 10% dialyzed FBS and 1X
CO:COLLECTION_SUMMARY            	penicillin/streptomycin, and incubated at 37°C for 30 min or 5 hours. After 30
CO:COLLECTION_SUMMARY            	min or 5 hours of isotope exposure, the medium was aspirated, and the cells were
CO:COLLECTION_SUMMARY            	washed 3 times with ice-cold PBS. Subsequently, metabolic activity was halted by
CO:COLLECTION_SUMMARY            	quenching with ice-cold acetonitrile (CH3CN), followed by the addition of
CO:COLLECTION_SUMMARY            	nanopure water (CH3CN: H2O at 2:1.5 (V/V)) to facilitate cell scraping and
CO:COLLECTION_SUMMARY            	collection.
CO:COLLECTION_PROTOCOL_FILENAME  	SIRM CELL CULTURE_Media-Cell Harvest_Protocol.pdf
CO:SAMPLE_TYPE                   	Cultured fibroblasts
CO:COLLECTION_METHOD             	Centrifuge and freeze
CO:STORAGE_CONDITIONS            	-80℃
#TREATMENT
TR:TREATMENT_SUMMARY             	A comparison of metabolite analysis was performed between wild-type NIH3T3 and
TR:TREATMENT_SUMMARY             	PRPS1-only expressing (PRPS2/PRPSAP1/PRPSAP2 knockout) NIH3T3 cells. CRISPR/Cas9
TR:TREATMENT_SUMMARY             	genome editing was performed as follows to generate the PRPS2/PRPSAP1/PRPSAP2 KO
TR:TREATMENT_SUMMARY             	clones: NIH3T3 cells were transduced with a dox-inducible lentiviral Cas9
TR:TREATMENT_SUMMARY             	plasmid (Addgene #50661) and selected with puromycin. Annealed crRNAs against
TR:TREATMENT_SUMMARY             	PRPSAP1 and PRPSAP2 were cloned into a lenti-sgRNA vector with hygromycin
TR:TREATMENT_SUMMARY             	selection (Addgene #104991) using BsmBI digestion. These lentiviral sgRNA
TR:TREATMENT_SUMMARY             	vectors were then introduced into cells expressing dox-inducible Cas9 and
TR:TREATMENT_SUMMARY             	selected with hygromycin. Cells were treated with 1 µg/mL doxycycline for a
TR:TREATMENT_SUMMARY             	week before clonally sorting them onto collagen-coated 96-well plates using BD
TR:TREATMENT_SUMMARY             	FACSAria. Clones were assayed for loss of protein expression by Western blotting
TR:TREATMENT_SUMMARY             	and were further checked for mutations by purifying genomic DNA and performing
TR:TREATMENT_SUMMARY             	PCR on the region spanning the edit site. To create PRPS2/PRPSAP1/PRPSAP2, PRPS2
TR:TREATMENT_SUMMARY             	crRNAs were first cloned into a lenti-sgRNA vector with neomycin selection
TR:TREATMENT_SUMMARY             	(Addgene #104992) using BsmBI digestion. These sgRNA vectors were introduced in
TR:TREATMENT_SUMMARY             	PRPSAP1/PRPSAP2 double KO cell lines to generate the PRPS2/PRPSAP1/PRPSAP2
TR:TREATMENT_SUMMARY             	triple KO cell lines. Cells were selected with neomycin and treated with 1
TR:TREATMENT_SUMMARY             	µg/mL doxycycline for a week before clonally sorting them onto collagen-coated
TR:TREATMENT_SUMMARY             	96-well plates using BD FACSAria. Clones were validated for loss of protein
TR:TREATMENT_SUMMARY             	expression by Western blotting. For the stable isotope experiment, cells were
TR:TREATMENT_SUMMARY             	either treated with glucose-free media containing 10 mM of 12C6-glucose or
TR:TREATMENT_SUMMARY             	13C6-glucose, supplemented with 10% dialyzed FBS and 1X penicillin/streptomycin,
TR:TREATMENT_SUMMARY             	and incubated at 37°C for 30 min or 5 hours before collection (described in
TR:TREATMENT_SUMMARY             	Collection summary).
#SAMPLEPREP
SP:SAMPLEPREP_SUMMARY            	Polar and non-polar metabolites were extracted using the solvent partition
SP:SAMPLEPREP_SUMMARY            	method – acetonitrile: water: chloroform (CH3CN: H2O: CHCl3 at 2:1.5:1 (V/V)).
SP:SAMPLEPREP_SUMMARY            	The aqueous phase containing polar metabolites and the organic phase containing
SP:SAMPLEPREP_SUMMARY            	non-polar metabolites were separated and dried using vacuum lyophilization
SP:SAMPLEPREP_SUMMARY            	(CentriVap Labconco) or a SpeedVac device. The protein pellets were washed in
SP:SAMPLEPREP_SUMMARY            	500 µL of 100% methanol and centrifuged at 12,000 x g at 4°C for 10 min. After
SP:SAMPLEPREP_SUMMARY            	discarding the supernatant, the protein pellet was dried in a SpeedVac
SP:SAMPLEPREP_SUMMARY            	centrifuge for 20 min. The resulting protein residue pellet was used for
SP:SAMPLEPREP_SUMMARY            	normalization.
SP:PROCESSING_STORAGE_CONDITIONS 	-80℃
SP:EXTRACT_STORAGE               	-80℃
#ANALYSIS
AN:ANALYSIS_TYPE                 	NMR
#NMR
NM:INSTRUMENT_NAME               	600MHz Bruker Avance III HD
NM:INSTRUMENT_TYPE               	FT-NMR
NM:NMR_EXPERIMENT_TYPE           	Other
NM:NMR_COMMENTS                  	NMR experiment type: 1D 1H-13C-HSQC
NM:SPECTROMETER_FREQUENCY        	600 MHz
NM:NMR_PROBE                     	5 mm Broad Band Observed (BBO) Prodigy probe
NM:NMR_SOLVENT                   	D2O
NM:NMR_TUBE_SIZE                 	3 mm
NM:SHIMMING_METHOD               	TopShim
NM:PULSE_SEQUENCE                	hsqcetgppgsisp2.2
NM:WATER_SUPPRESSION             	25Hz bandwidth
NM:RECEIVER_GAIN                 	90.5
NM:NUMBER_OF_SCANS               	1024
NM:DUMMY_SCANS                   	16
NM:ACQUISITION_TIME              	2
NM:SPECTRAL_WIDTH                	15 ppm
NM:NUM_DATA_POINTS_ACQUIRED      	16000
NM:LINE_BROADENING               	4 Hz
NM:APODIZATION                   	Exponential
NM:BASELINE_CORRECTION_METHOD    	Bernstein polynomial fit
NM:CHEMICAL_SHIFT_REF_STD        	0.00 ppm
#NMR_METABOLITE_DATA
NMR_METABOLITE_DATA:UNITS	nmoles/mg dry residue protein
NMR_METABOLITE_DATA_START
Samples	Cunninghan_UC_PRPS_Cells_110222.101	Cunninghan_UC_PRPS_Cells_110222.102	Cunninghan_UC_PRPS_Cells_110222.103	Cunninghan_UC_PRPS_Cells_110222.104	Cunninghan_UC_PRPS_Cells_110222.105	Cunninghan_UC_PRPS_Cells_110222.106	Cunninghan_UC_PRPS_Cells_110222.107	Cunninghan_UC_PRPS_Cells_110222.108	Cunninghan_UC_PRPS_Cells_110222.109	Cunninghan_UC_PRPS_Cells_110222.110	Cunninghan_UC_PRPS_Cells_110222.111	Cunninghan_UC_PRPS_Cells_110222.112	Cunninghan_UC_PRPS_Cells_110222.113	Cunninghan_UC_PRPS_Cells_110222.114	Cunninghan_UC_PRPS_Cells_110222.115	Cunninghan_UC_PRPS_Cells_110222.116	Cunninghan_UC_PRPS_Cells_110222.117	Cunninghan_UC_PRPS_Cells_110222.118	Cunninghan_UC_PRPS_Cells_110222.201	Cunninghan_UC_PRPS_Cells_110222.202	Cunninghan_UC_PRPS_Cells_110222.203	Cunninghan_UC_PRPS_Cells_110222.204	Cunninghan_UC_PRPS_Cells_110222.205	Cunninghan_UC_PRPS_Cells_110222.206	Cunninghan_UC_PRPS_Cells_110222.207	Cunninghan_UC_PRPS_Cells_110222.208	Cunninghan_UC_PRPS_Cells_110222.209	Cunninghan_UC_PRPS_Cells_110222.210	Cunninghan_UC_PRPS_Cells_110222.211	Cunninghan_UC_PRPS_Cells_110222.212	Cunninghan_UC_PRPS_Cells_110222.213	Cunninghan_UC_PRPS_Cells_110222.214	Cunninghan_UC_PRPS_Cells_110222.215	Cunninghan_UC_PRPS_Cells_110222.216	Cunninghan_UC_PRPS_Cells_110222.217	Cunninghan_UC_PRPS_Cells_110222.218
Factors	Sample source:NIH3T3 | Genotype:Wildtype1_Gr1 | Treatment:U12C glucose | Treatment duration:30 mins	Sample source:NIH3T3 | Genotype:Wildtype2_Gr1 | Treatment:U12C glucose | Treatment duration:30 mins	Sample source:NIH3T3 | Genotype:Wildtype3_Gr1 | Treatment:U12C glucose | Treatment duration:30 mins	Sample source:NIH3T3 | Genotype:Wildtype1_Gr2 | Treatment:U13C glucose | Treatment duration:30 mins	Sample source:NIH3T3 | Genotype:Wildtype2_Gr2 | Treatment:U13C glucose | Treatment duration:30 mins	Sample source:NIH3T3 | Genotype:Wildtype3_Gr2 | Treatment:U13C glucose | Treatment duration:30 mins	Sample source:NIH3T3 | Genotype:Wildtype1-Gr3 | Treatment:U13C glucose | Treatment duration:5 hrs	Sample source:NIH3T3 | Genotype:Wildtype2-Gr3 | Treatment:U13C glucose | Treatment duration:5 hrs	Sample source:NIH3T3 | Genotype:Wildtype3-Gr3 | Treatment:U13C glucose | Treatment duration:5 hrs	Sample source:NIH3T3 | Genotype:P2/AP1/AP2 KO1_Gr4 | Treatment:U12C glucose | Treatment duration:30 mins	Sample source:NIH3T3 | Genotype:P2/AP1/AP2 KO2_Gr4 | Treatment:U12C glucose | Treatment duration:30 mins	Sample source:NIH3T3 | Genotype:P2/AP1/AP2 KO3_Gr4 | Treatment:U12C glucose | Treatment duration:30 mins	Sample source:NIH3T3 | Genotype:P2/AP1/AP2 KO1_Gr5 | Treatment:U13C glucose | Treatment duration:30 mins	Sample source:NIH3T3 | Genotype:P2/AP1/AP2 KO2_Gr5 | Treatment:U13C glucose | Treatment duration:30 mins	Sample source:NIH3T3 | Genotype:P2/AP1/AP2 KO3_Gr5 | Treatment:U13C glucose | Treatment duration:30 mins	Sample source:NIH3T3 | Genotype:P2/AP1/AP2 KO1_Gr6 | Treatment:U13C glucose | Treatment duration:5 hrs	Sample source:NIH3T3 | Genotype:P2/AP1/AP2 KO2_Gr6 | Treatment:U13C glucose | Treatment duration:5 hrs	Sample source:NIH3T3 | Genotype:P2/AP1/AP2 KO3_Gr6 | Treatment:U13C glucose | Treatment duration:5 hrs	Sample source:NIH3T3 | Genotype:Wildtype1_Gr1 | Treatment:U12C glucose | Treatment duration:30 mins	Sample source:NIH3T3 | Genotype:Wildtype2_Gr1 | Treatment:U12C glucose | Treatment duration:30 mins	Sample source:NIH3T3 | Genotype:Wildtype3_Gr1 | Treatment:U12C glucose | Treatment duration:30 mins	Sample source:NIH3T3 | Genotype:Wildtype1_Gr2 | Treatment:U13C glucose | Treatment duration:30 mins	Sample source:NIH3T3 | Genotype:Wildtype2_Gr2 | Treatment:U13C glucose | Treatment duration:30 mins	Sample source:NIH3T3 | Genotype:Wildtype3_Gr2 | Treatment:U13C glucose | Treatment duration:30 mins	Sample source:NIH3T3 | Genotype:Wildtype1-Gr3 | Treatment:U13C glucose | Treatment duration:5 hrs	Sample source:NIH3T3 | Genotype:Wildtype2-Gr3 | Treatment:U13C glucose | Treatment duration:5 hrs	Sample source:NIH3T3 | Genotype:Wildtype3-Gr3 | Treatment:U13C glucose | Treatment duration:5 hrs	Sample source:NIH3T3 | Genotype:P2/AP1/AP2 KO1_Gr4 | Treatment:U12C glucose | Treatment duration:30 mins	Sample source:NIH3T3 | Genotype:P2/AP1/AP2 KO2_Gr4 | Treatment:U12C glucose | Treatment duration:30 mins	Sample source:NIH3T3 | Genotype:P2/AP1/AP2 KO3_Gr4 | Treatment:U12C glucose | Treatment duration:30 mins	Sample source:NIH3T3 | Genotype:P2/AP1/AP2 KO1_Gr5 | Treatment:U13C glucose | Treatment duration:30 mins	Sample source:NIH3T3 | Genotype:P2/AP1/AP2 KO2_Gr5 | Treatment:U13C glucose | Treatment duration:30 mins	Sample source:NIH3T3 | Genotype:P2/AP1/AP2 KO3_Gr5 | Treatment:U13C glucose | Treatment duration:30 mins	Sample source:NIH3T3 | Genotype:P2/AP1/AP2 KO1_Gr6 | Treatment:U13C glucose | Treatment duration:5 hrs	Sample source:NIH3T3 | Genotype:P2/AP1/AP2 KO2_Gr6 | Treatment:U13C glucose | Treatment duration:5 hrs	Sample source:NIH3T3 | Genotype:P2/AP1/AP2 KO3_Gr6 | Treatment:U13C glucose | Treatment duration:5 hrs
Acetic acid (Ace-2_1H)	3.533742377	1.046596039	0.947180137	1.03508121	0.985056439	1.226731889	0.859671822	1.660304802	1.09214541	0.992543781	0.767085585	0.651602284	0.746269507	2.17236328	0.87546177	0.779979124	1.39070224	1.242190483	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0
Glucose (aGlc-1_1H)	34.42731015	35.67643746	29.36768519	1.151968772	1.745857444	2.922294331	0	1.642768274	0	23.39564859	21.40825327	20.26856742	0	0	1.561944195	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0
Glucose (aGlc-1 13C-A_1H)	0	0	0	30.36755424	45.54777891	27.25526768	17.72182299	22.82879466	21.6154734	0	0	0	21.51740354	17.81076451	17.36484905	10.30467765	8.454911246	14.68233168	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0
Alanine (Ala-3_1H)	7.465952859	7.369005386	7.502743186	3.403157393	3.896205183	3.196834882	1.935897749	1.897009288	2.004635216	14.85855601	12.99907708	13.4377266	3.788642889	3.54019062	4.608561487	2.788785291	3.220779185	2.955180726	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0
Alanine (Ala13C-B_1H)	0	0	0	2.900161236	3.234932772	4.498079583	3.321784553	3.24229587	2.296994813	0	0	0	8.175092214	7.349023506	9.532964213	14.10155221	16.88406948	11.96135798	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0
AMP (AMP-3'_1H)	27.69860492	19.95689938	22.00008411	25.53644859	15.48135577	16.80497952	13.19476369	12.48683784	16.75917852	21.13341265	18.08454811	16.53691988	16.11116366	10.70298796	13.33861066	11.73358623	11.6772167	20.94835735	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0
Aspartic acid (Asp-3_1H)	21.0971149	18.90689107	19.72958784	18.34565353	18.85762508	18.17994909	0	0	0	23.06703323	19.70931674	17.62078057	18.41187616	16.45622839	15.91037219	2.281825014	3.215795296	2.562447786	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0
ATP (ATP-3'13C-A_1H)	0	0	0	0	0	0	43.07653181	34.99622935	33.69724732	0	0	0	0	0	0	68.54166395	74.22552123	71.35409548	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0
ATP (ATP-4'_1H)	64.74110388	60.59983678	64.05143167	22.88068248	45.63511041	37.23858889	17.03977915	42.32481323	36.77979655	53.86576652	43.01344257	48.51584857	24.60003154	22.51443986	18.12827351	10.53994693	29.46658288	38.38177071	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0
AXP (AXP-1'_1H)	38.43466455	34.13692002	33.76151231	30.0563685	29.96634967	26.39862537	20.16357834	21.40629407	21.62313527	33.99869899	29.70356353	27.73206657	25.30103254	22.50749302	21.05659025	20.1963615	22.27164391	26.22676519	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0
AXP (AXP-1' 13C-B_1H)	0	0	0	1.417325825	1.540782348	2.065599307	6.384353663	5.355029234	5.392339633	0	0	0	1.195498633	1.344797682	2.827285875	6.484764275	5.937388752	6.38642371	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0
AXP (AXP-2_1H)	25.87717815	23.51211262	23.7966203	21.68915882	23.90561892	19.0021547	19.37754019	19.58870221	18.69372671	21.50357541	21.18097906	18.41020162	18.51091096	17.19483841	14.5029149	13.61404216	16.14664885	20.65719737	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0
AXP (AXP-8_1H)	43.24648277	37.63637504	37.9589774	34.36606104	34.56240933	30.3418931	26.5965277	28.26470608	28.61474662	34.01129947	30.94211341	28.69130479	30.14900808	26.33349075	24.5288231	23.35094083	28.18904287	31.53224293	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0
Choline (Choline_1H)	0.580688642	0.380760946	0.072533274	0.149876253	0.251603719	0	0	0	0.298223255	1.752560008	1.610933789	1.726296161	1.600260029	1.494536413	1.529009165	2.414759337	2.577689107	2.545736031	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0
Creatine (Creatine-A_1H)	28.04390014	24.47466632	25.21687188	22.31299771	23.55779115	20.34107134	16.10832628	17.0150881	16.6635682	38.47055979	34.76818085	34.57614157	33.47384835	29.39804733	28.87571071	21.60421668	25.6247148	24.55831876	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0
Creatinine (Creatinine-P-A_1H)	2.697323689	2.369567417	2.587032553	2.204367339	2.752037213	2.771098727	2.097707136	2.796975455	2.582937597	8.103731961	7.246395254	8.651918389	7.052039903	6.875352114	5.827216762	7.059073796	7.755934071	9.11783877	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0
Cysteine-Glutathione (Cys-GSH_1H)	48.83494543	43.79912372	45.24760317	41.18316261	40.20383787	37.01509315	27.64983446	28.11284674	28.62398534	50.61393568	43.23194835	46.34721937	42.08826566	40.36476441	41.27473586	29.20426728	30.56872139	30.39826651	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0
Formic acid (Formate_1H)	6.374396695	5.932521071	5.829330038	5.807726677	5.884316054	5.794486114	6.314384117	5.382695795	5.966131525	5.888598745	5.984190697	6.579892836	6.119741968	6.066496579	5.393311051	5.223222087	6.486807212	6.649391642	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0
Fumaric acid (Fumarate_1H)	0.667950711	0.633320251	0.665445203	0.514726426	0.524993282	0.532079095	0.066456378	0	0	0.887395561	0.856628644	0.686385069	0.712299098	0.73716036	0.698622987	0.185334113	0.168067541	0.239239586	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0
Glutamic acid (Glu-4_1H)	188.7150712	169.7794671	177.4897587	140.6479516	153.5111207	149.2182901	20.64738674	22.24430028	25.49135422	174.6049591	158.4563253	156.5452798	152.7967818	146.6392892	140.0196125	37.69337168	42.43188602	48.50336501	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0
Glycine (Gly_1H)	116.9775679	104.318311	107.0696273	98.78706797	101.8416685	91.27771111	128.313408	132.4497377	124.1755176	73.11906677	65.55173279	72.54430115	65.07590281	61.52761006	72.05321276	90.17116747	98.52300834	101.5607871	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0
GpC (GPC_1H)	3.785890832	4.588264363	3.766633552	1.397095744	1.636899511	2.141528192	13.52765537	15.69634367	13.20730402	2.530070941	2.107087666	2.242938001	1.760983701	1.380541306	1.374466436	16.13741012	16.15490653	17.27548703	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0
Glutathione (GSH-8_1H)	39.57885474	29.39738646	32.26830016	30.3017352	25.16623714	26.24700906	16.80498082	16.68840185	17.65900267	35.33139037	30.05047013	33.84009047	30.7025606	27.56554344	28.78183692	20.00445309	22.56869423	23.79673285	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0
GTP (GTP_1H)	4.096967735	3.436857399	4.312665493	3.475142754	3.234594188	3.171302018	2.964448802	2.837970065	3.120563488	6.111369847	6.513371402	6.003976696	5.115606608	5.577744858	5.103449267	3.040047936	5.19510927	4.368949436	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0
GTP (GTP-1' 13C-A_1H)	0	0	0	0	0	0	4.331977388	3.268681252	2.513484019	0	0	0	0	0	0	1.817516215	0.998845828	2.121461026	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0
Histidine (His-5_1H)	6.692111071	5.61283387	6.310544348	7.254259973	6.948610778	6.18489855	8.845695823	8.717629647	5.722664255	7.377432672	6.667219821	7.491756147	6.975718224	7.184542633	7.905695726	6.361753874	6.564798527	6.71496327	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0
Isoleucine (Ile-B_1H)	24.95805191	21.93464528	22.79189017	21.26670944	22.20369413	19.55161878	19.21665812	19.57425466	19.52758498	14.57315091	12.45762357	13.02730919	12.70626044	11.9560061	12.07810898	15.39249633	16.4323932	18.74016334	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0
Inosine (Inosine_1H)	2.790858772	1.991693477	1.917045793	1.365985006	1.478117265	1.874969949	5.355187014	5.298134695	5.540670735	2.168908537	2.042023174	2.155682796	1.684616928	2.177610538	1.863097658	3.489443084	4.206118988	5.183095086	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0
Lactic acid (Lac-2_1H)	21.37667337	19.15307038	19.80957776	0	0	0	0	0	0	26.20155618	24.48834372	23.24737188	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0
Lactic acid/Threonine (Lac-3/Thr_1H)	147.2327121	123.7208163	131.5485692	94.35872484	100.5939926	90.92206937	97.49395339	96.69750406	95.71444925	115.661503	100.8064805	104.0338623	65.4987875	62.87341527	65.29728731	97.93993276	107.3852596	112.0603358	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0
Lactic acid (Lac13C-A_1H)	0	0	0	30.2627451	30.05942685	29.06001725	28.44803046	29.0005104	27.17078548	0	0	0	39.44163152	34.26038588	36.33635352	31.28765492	37.01582654	41.74710057	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0
Leucine (Leu_1H)	40.88652749	33.09315887	36.1274015	32.70805505	36.43303409	32.4717186	27.78737347	32.64323336	30.61954094	24.83442462	19.96703662	23.81789333	20.28278873	20.04588216	18.19100027	23.60056065	25.96314107	25.5895483	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0
Myo-Inositol (m-Ins-2_1H)	54.89014083	50.8758556	51.10130469	51.08494222	53.59825478	47.40770784	55.48637987	57.61159377	59.92512044	111.2022529	98.54957506	103.797298	101.4289001	99.06456333	101.2525392	114.0402747	127.2322884	131.0492741	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0
Methionine (Met_1H)	14.21443909	12.03067173	12.78554924	12.00412872	11.70022918	11.52145215	10.31623767	11.11578581	10.82717465	8.68222022	7.296436856	7.815124144	7.010059721	6.835954193	6.672747927	7.803388368	8.873437062	9.601274096	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0
NAD (NAD+-A1'_1H)	6.159498534	4.686625848	4.463742054	4.916605847	4.617343345	4.943916527	4.853650385	4.506448176	5.107775035	4.756207999	4.609237343	4.448610942	4.848952715	4.511644636	4.606929016	4.69054645	4.849515054	3.936859037	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0
NAD (NAD+-N1'_1H)	6.19128565	5.113882032	5.323966661	5.453950648	4.792432816	4.921445863	4.373360154	2.798845478	3.288580194	4.939485541	5.090590255	5.060674888	5.093946028	4.545169065	4.618340854	3.030700972	3.113457403	3.679326667	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0
NAD (NAD+-N1' 13C-B_1H)	0	0	0	0	0	0	1.116446484	1.598421158	1.533164719	0	0	0	0	0	0	1.108336803	1.463505169	1.737300862	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0
NAD (NAD+-N2_1H)	5.887600972	5.355181809	4.900516518	5.443491218	4.477476571	4.723294083	5.191387873	4.227246535	5.059365564	4.557296935	4.695592907	5.187351475	4.43851265	4.536730158	4.622338705	4.913978408	4.854892069	5.204014778	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0
NAD (NAD+-N6_1H)	6.437816593	5.597940657	4.667641987	5.446539493	4.665934549	4.800649773	5.488931339	4.396846895	5.338208388	4.29969773	4.47552615	5.04922957	4.810693502	4.626827364	4.589244038	4.861601339	5.103070327	5.50748116	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0
NADP (NADP+-N2_1H)	0.921552527	0.695514489	0.612793372	0.541718866	0.575847711	0.552052414	0.544201194	0.724216218	0.511824023	0.836838058	0.787877513	0.941066885	0.646494999	0.600190997	0.58421852	0.598231524	0.665478825	0.703856664	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0
NADP (NADP+-N6_1H)	0.814186384	0.790224423	0.693385066	0.631382649	0.687172903	0.492355063	0.750593407	0.3475004	0.529968104	0.736283312	0.939824511	0.947396147	0.700098484	0.63355285	0.689595453	0.503013662	0.742263323	0.903953456	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0
Choline phosphate (P-Choline_1H)	100.436498	88.41885113	89.41813059	85.81573296	87.07020873	82.65568613	109.331121	115.0267934	113.1959622	68.52000711	55.85101134	55.46300696	54.92402669	47.57518187	45.54323969	62.04791487	65.76308426	72.81875028	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0
Phenylalanine (Phe-3,5_1H)	15.53508301	13.46743743	13.74883576	13.27379422	13.23367788	13.16975844	12.07176721	12.44133812	12.56832114	8.646064453	7.517019823	8.038741595	7.635425139	7.402280513	6.83109322	8.620813852	9.291881545	10.38056126	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0
Serine (Ser-2_1H)	85.11621845	65.87384915	65.78449216	60.56837361	60.91618284	60.56898206	41.84303357	45.65015616	45.87486552	70.1490957	40.44857199	50.87022263	41.35688144	35.42093628	46.36206497	59.22519838	66.3705519	73.09653397	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0
Serine (Ser-3_1H)	95.0889689	77.78164042	78.42468314	67.54264029	67.40282022	66.1475344	51.56754813	56.49737569	54.83548969	85.70248603	60.51974607	68.82958575	58.94281089	63.39228585	62.8650436	76.39399595	96.61122803	108.8179008	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0
Taurine (Tau-2 (NCH2)_1H)	33.1063857	33.41204882	35.2354315	35.66336695	25.04965436	32.68230059	29.34820772	30.91838762	34.47322335	68.14873257	61.51470145	60.99399746	59.15705302	59.00821292	57.75172557	52.81567824	62.7091398	61.47287076	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0
Threonine (Thr_1H)	95.93576641	84.07622442	86.53384979	76.64613294	74.03932141	72.14878153	79.81165941	78.23468322	77.31129783	63.63356655	61.1202743	64.87719047	56.7240055	50.16337805	53.59879724	74.38499609	89.27011803	91.11869612	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0
Tryptophan (Trp-4_1H)	1.815284481	1.689474208	2.004749119	1.497931745	1.306356864	1.273925344	1.884806393	1.601499132	1.523860766	1.228012636	0.715907121	1.134146894	0.975092235	0.875860202	1.070126376	1.125522637	1.30309015	1.153676011	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0
Tyrosine (Tyr-3,5_1H)	13.92620382	11.55218586	12.00702786	11.85269689	11.46237036	11.03934717	10.41395212	10.29434352	9.813529896	7.385335037	6.330554296	6.647543506	6.039786228	6.127449185	6.683530502	7.586544993	8.597154297	8.798459633	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0
UDP-N-acetylgalactosamine (UDP-GalNAc_1H)	1.927637234	1.755994415	2.374781545	3.115599442	2.534137981	3.161644346	1.295864554	1.783942154	1.362351716	1.788076054	2.020382449	1.697266971	1.982258127	2.309736968	2.350689339	0.549372041	1.093802406	1.064073189	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0
UDP-glucose (UDP-Glc_1H)	4.564957729	3.014947932	4.079350548	3.631731387	2.391283332	3.236027758	3.64731311	3.228765973	4.060469557	3.574412006	3.924453372	3.955541098	2.843257927	2.63431528	2.360396914	2.080594805	2.244083851	3.801851206	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0
UXP (UXP-1'_1H)	19.57214216	21.83125552	20.06571114	14.10681278	16.96195135	14.92186407	9.843380047	11.76101528	11.77854381	19.85152451	19.65308545	16.76359922	13.49633849	13.42625346	13.08944564	11.96527884	14.07048289	17.02614972	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0
UXP (UXP-1' 13C-A_1H)	0	0	0	2.466018463	3.246292807	4.810104019	8.221718409	7.553450932	7.427993635	0	0	0	0.933611961	0	0	3.872547455	3.646378426	4.470535552	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0
UXP (UXP-5'_1H)	10.48595631	12.57403664	10.45288528	9.276734761	13.11644074	10.64717285	10.97023156	11.64115405	11.07260188	12.40677799	10.07994351	11.39740885	17.33174807	12.55299602	13.40298432	12.68710244	11.89837646	13.61468654	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0
Valine (Val-B_1H)	28.52711743	24.89198227	25.92825044	23.93716258	25.3171037	23.88339231	23.59130433	24.36649867	24.0386357	15.96870398	12.69594033	14.15424786	13.59462877	12.54765574	12.96457197	18.33597307	20.69496715	21.50544448	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0
Glucose (aGlc-1_1D 1H-13C-HSQC)	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0.0000	0.0000	0.0000	66.5285	104.3786	61.5717	50.0267	56.5482	49.8884	0.0000	0.0000	0.0000	49.7330	44.0631	49.4725	31.1821	31.6429	42.4131
Alanine (Ala-3_1D 1H-13C-HSQC)	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0.0000	0.0000	0.0000	3.4542	2.9479	4.3396	4.2468	4.3334	3.9809	0.0000	0.0000	0.0000	11.3091	8.9883	11.5781	16.7340	19.4470	15.2584
AMP (AMP-3'_1D 1H-13C-HSQC)	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0.0000	0.0000	0.0000	1.6772	3.9215	3.3595	5.8197	5.9523	4.9297	0.0000	0.0000	0.0000	4.2122	3.5956	6.3065	6.3552	8.4637	8.8279
ATP (ATP-4'_1D 1H-13C-HSQC)	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0.0000	0.0000	0.0000	27.7461	25.2230	28.0168	36.9417	37.5162	40.2957	0.0000	0.0000	0.0000	24.0661	18.5072	22.5337	24.3691	27.0829	33.6669
AXP (AXP-1'_1D 1H-13C-HSQC)	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0.0000	0.0000	0.0000	4.4243	7.1863	4.6774	16.1914	14.2798	15.4487	0.0000	0.0000	0.0000	6.4234	5.3575	10.1091	11.7223	16.6044	16.0078
Glutamic acid (Glu-3a_1D 1H-13C-HSQC)	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0.0000	0.0000	0.0000	3.7830	2.9650	2.7620	22.2602	17.3600	17.2332	0.0000	0.0000	0.0000	0.9630	1.3737	2.1902	5.3162	7.5301	8.2330
Glutamic acid (Glu-4_1D 1H-13C-HSQC)	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0.0000	0.0000	0.0000	19.4596	20.3483	21.1218	44.7103	42.3943	42.6828	0.0000	0.0000	0.0000	5.7659	3.7659	5.4759	6.9841	8.1725	7.3969
Glutathione (GSH-4_1D 1H-13C-HSQC)	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0.0000	0.0000	0.0000	0.0000	0.0000	2.1916	10.7708	10.4450	12.0994	0.0000	0.0000	0.0000	0.0000	0.0000	0.0000	0.0000	0.0000	0.0000
GTP (GTP_1D 1H-13C-HSQC)	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0.0000	0.0000	0.0000	0.0000	0.0000	2.0017	10.2148	13.8145	11.5821	0.0000	0.0000	0.0000	0.0000	0.0000	0.0000	4.8950	6.2845	7.9080
Lactic acid (Lac-2_1D 1H-13C-HSQC)	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0.0000	0.0000	0.0000	23.1747	20.4504	21.5558	12.6148	10.1225	11.0839	0.0000	0.0000	0.0000	27.5430	24.9376	24.2287	12.8827	17.0691	21.2698
Lactic acid/Threonine (Lac-3/Thr_1D 1H-13C-HSQC)	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0.0000	0.0000	0.0000	30.2627	30.0594	29.0600	28.4480	29.0005	27.1708	0.0000	0.0000	0.0000	39.4416	34.2604	36.3364	31.2877	37.0158	41.7471
NAD (NAD+-N1'_1D 1H-13C-HSQC)	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0.0000	0.0000	0.0000	1.3167	1.0801	1.2807	7.9011	5.7055	8.1748	0.0000	0.0000	0.0000	0.0000	0.0000	0.0000	6.9524	8.0044	8.5272
UXP (UXP-1'_1D 1H-13C-HSQC)	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0.0000	0.0000	0.0000	10.7525	12.4008	8.5492	24.6535	19.6603	22.4862	0.0000	0.0000	0.0000	7.3828	5.9740	7.1399	16.7036	17.2374	19.6746
NMR_METABOLITE_DATA_END
#METABOLITES
METABOLITES_START
metabolite_name	Standardized name	Super class	Main class	Sub class	Pubchem CID	CHEBI_ID	HMDB_ID	LM_ID	KEGG_ID	INCHI_KEY	REFMET_ID
Acetic acid (Ace-2_1H)	Acetic acid	Fatty Acyls	Fatty acids	Saturated FA	176	15366	HMDB0000042	LMFA01010002	C00033	QTBSBXVTEAMEQO-UHFFFAOYSA-N	RM0153572
Glucose (aGlc-1_1H)	Glucose	Carbohydrates	Monosaccharides	Hexoses	5793	4167	HMDB0304632		C00031	WQZGKKKJIJFFOK-GASJEMHNSA-N	RM0135901
Glucose (aGlc-1 13C-A_1H)	Glucose	Carbohydrates	Monosaccharides	Hexoses	5793	4167	HMDB0304632		C00031	WQZGKKKJIJFFOK-GASJEMHNSA-N	RM0135901
Alanine (Ala-3_1H)	Alanine	Organic acids	Amino acids and peptides	Amino acids	5950	16977	HMDB0000161		C00041	QNAYBMKLOCPYGJ-REOHCLBHSA-N	RM0135898
Alanine (Ala13C-B_1H)	Alanine	Organic acids	Amino acids and peptides	Amino acids	5950	16977	HMDB0000161		C00041	QNAYBMKLOCPYGJ-REOHCLBHSA-N	RM0135898
AMP (AMP-3'_1H)	AMP	Nucleic acids	Purines	Purine rNMP	6083	16027	HMDB0000045		C00020	UDMBCSSLTHHNCD-KQYNXXCUSA-N	RM0138921
Aspartic acid (Asp-3_1H)	Aspartic acid	Organic acids	Amino acids and peptides	Amino acids	5960	17053	HMDB0000191		C00049	CKLJMWTZIZZHCS-REOHCLBHSA-N	RM0135905
ATP (ATP-3'13C-A_1H)	ATP	Nucleic acids	Purines	Purine rNTP	5957	15422	HMDB0000538		C00002	ZKHQWZAMYRWXGA-KQYNXXCUSA-N	RM0138930
ATP (ATP-4'_1H)	ATP	Nucleic acids	Purines	Purine rNTP	5957	15422	HMDB0000538		C00002	ZKHQWZAMYRWXGA-KQYNXXCUSA-N	RM0138930
AXP (AXP-1'_1H)	AXP	Nucleic acids	Purines	Purine rNXP
AXP (AXP-1' 13C-B_1H)	AXP	Nucleic acids	Purines	Purine rNXP
AXP (AXP-2_1H)	AXP	Nucleic acids	Purines	Purine rNXP
AXP (AXP-8_1H)	AXP	Nucleic acids	Purines	Purine rNXP
Choline (Choline_1H)	Choline	Organic nitrogen compounds	Cholines	Cholines	305	15354	HMDB0000097		C00114	OEYIOHPDSNJKLS-UHFFFAOYSA-N	RM0028739
Creatine (Creatine-A_1H)	Creatine	Organic acids	Amino acids and peptides	Amino acids	586	16919	HMDB0000064		C00300	CVSVTCORWBXHQV-UHFFFAOYSA-N	RM0135876
Creatinine (Creatinine-P-A_1H)	Creatinine	Organoheterocyclic compounds	Azoles	Imidazolines	137319715	16737	HMDB0000562		C00791	DDRJAANPRJIHGJ-UHFFFAOYSA-N	RM0028430
Cysteine-Glutathione (Cys-GSH_1H)	Cysteine-Glutathione	Organic acids	Amino acids and peptides
Formic acid (Formate_1H)	Formic acid	Organic acids	Carboxylic acids	Carboxylic acids	284	30751	HMDB0000142		C00058	BDAGIHXWWSANSR-UHFFFAOYSA-N	RM0135892
Fumaric acid (Fumarate_1H)	Fumaric acid	Organic acids	TCA acids	TCA acids	444972	18012	HMDB0000134		C00122	VZCYOOQTPOCHFL-OWOJBTEDSA-N	RM0135891
Glutamic acid (Glu-4_1H)	Glutamic acid	Organic acids	Amino acids and peptides	Amino acids	33032	16015	HMDB0000148		C00025	WHUUTDBJXJRKMK-VKHMYHEASA-N	RM0135893
Glycine (Gly_1H)	Glycine	Organic acids	Amino acids and peptides	Amino acids	750	15428	HMDB0000123		C00037	DHMQDGOQFOQNFH-UHFFFAOYSA-N	RM0135888
GpC (GPC_1H)	GpC	Nucleic acids	Dinucleotides	Diribonucleotides	145459099	136671				DQTQQHQGSZWEMR-VMIOUTBZSA-N	RM0049797
Glutathione (GSH-8_1H)	Glutathione	Organic acids	Amino acids and peptides	Tripeptides	124886	16856	HMDB0000125		C00051	RWSXRVCMGQZWBV-WDSKDSINSA-N	RM0135889
GTP (GTP_1H)	GTP	Nucleic acids	Purines	Purine rNTP	135398633	15996	HMDB0001273		C00044	XKMLYUALXHKNFT-UUOKFMHZSA-N	RM0136084
GTP (GTP-1' 13C-A_1H)	GTP	Nucleic acids	Purines	Purine rNTP	135398633	15996	HMDB0001273		C00044	XKMLYUALXHKNFT-UUOKFMHZSA-N	RM0136084
Histidine (His-5_1H)	Histidine	Organic acids	Amino acids and peptides	Amino acids	6274	15971	HMDB0000177		C00135	HNDVDQJCIGZPNO-YFKPBYRVSA-N	RM0129894
Isoleucine (Ile-B_1H)	Isoleucine	Organic acids	Amino acids and peptides	Amino acids	6306	17191	HMDB0000172	LMFA01100047	C00407	AGPKZVBTJJNPAG-WHFBIAKZSA-N	RM0134975
Inosine (Inosine_1H)	Inosine	Nucleic acids	Purines	Purine ribonucleosides	135398641	17596	HMDB0000195		C00294	UGQMRVRMYYASKQ-KQYNXXCUSA-N	RM0135908
Lactic acid (Lac-2_1H)	Lactic acid	Organic acids	Short-chain acids	Short-chain acids	107689	422	HMDB0000190		C00186	JVTAAEKCZFNVCJ-REOHCLBHSA-N	RM0135904
Lactic acid/Threonine (Lac-3/Thr_1H)	Lactic acid/Threonine	Organic acids
Lactic acid (Lac13C-A_1H)	Lactic acid	Organic acids	Short-chain acids	Short-chain acids	107689	422	HMDB0000190		C00186	JVTAAEKCZFNVCJ-REOHCLBHSA-N	RM0135904
Leucine (Leu_1H)	Leucine	Organic acids	Amino acids and peptides	Amino acids	6106	15603	HMDB0000687		C00123	ROHFNLRQFUQHCH-YFKPBYRVSA-N	RM0136379
Myo-Inositol (m-Ins-2_1H)	Inositol	Organic oxygen compounds	Alcohols and polyols	Inositols							RM0004841
Methionine (Met_1H)	Methionine	Organic acids	Amino acids and peptides	Amino acids	6137	16643	HMDB0000696		C00073	FFEARJCKVFRZRR-BYPYZUCNSA-N	RM0161726
NAD (NAD+-A1'_1H)	NAD	Nucleic acids	Nicotinamides	Nicotinamide dinucleotides	5892	44215	HMDB0000902		C00003	BAWFJGJZGIEFAR-NNYOXOHSSA-N	RM0133217
NAD (NAD+-N1'_1H)	NAD	Nucleic acids	Nicotinamides	Nicotinamide dinucleotides	5892	44215	HMDB0000902		C00003	BAWFJGJZGIEFAR-NNYOXOHSSA-N	RM0133217
NAD (NAD+-N1' 13C-B_1H)	NAD	Nucleic acids	Nicotinamides	Nicotinamide dinucleotides	5892	44215	HMDB0000902		C00003	BAWFJGJZGIEFAR-NNYOXOHSSA-N	RM0133217
NAD (NAD+-N2_1H)	NAD	Nucleic acids	Nicotinamides	Nicotinamide dinucleotides	5892	44215	HMDB0000902		C00003	BAWFJGJZGIEFAR-NNYOXOHSSA-N	RM0133217
NAD (NAD+-N6_1H)	NAD	Nucleic acids	Nicotinamides	Nicotinamide dinucleotides	5892	44215	HMDB0000902		C00003	BAWFJGJZGIEFAR-NNYOXOHSSA-N	RM0133217
NADP (NADP+-N2_1H)	NADP	Nucleic acids	Nicotinamides	Nicotinamide dinucleotides	5885	44409	HMDB0304435		C00006	XJLXINKUBYWONI-NNYOXOHSSA-N	RM0133216
NADP (NADP+-N6_1H)	NADP	Nucleic acids	Nicotinamides	Nicotinamide dinucleotides	5885	44409	HMDB0304435		C00006	XJLXINKUBYWONI-NNYOXOHSSA-N	RM0133216
Choline phosphate (P-Choline_1H)	Choline phosphate	Organic nitrogen compounds	Cholines	Cholines	135437	18132	HMDB0001565			YHHSONZFOIEMCP-UHFFFAOYSA-N	RM0007522
Phenylalanine (Phe-3,5_1H)	Phenylalanine	Organic acids	Amino acids and peptides	Amino acids	6140	17295	HMDB0000159		C00079	COLNVLDHVKWLRT-QMMMGPOBSA-N	RM0135897
Serine (Ser-2_1H)	Serine	Organic acids	Amino acids and peptides	Amino acids	5951	17115	HMDB0000187		C00065	MTCFGRXMJLQNBG-REOHCLBHSA-N	RM0032433
Serine (Ser-3_1H)	Serine	Organic acids	Amino acids and peptides	Amino acids	5951	17115	HMDB0000187		C00065	MTCFGRXMJLQNBG-REOHCLBHSA-N	RM0032433
Taurine (Tau-2 (NCH2)_1H)	Taurine	Organic acids	Sulfonic acids	Sulfonic acids	1123	15891	HMDB0000251		C00245	XOAAWQZATWQOTB-UHFFFAOYSA-N	RM0118223
Threonine (Thr_1H)	Threonine	Organic acids	Amino acids and peptides	Amino acids	6288	16857	HMDB0000167		C00188	AYFVYJQAPQTCCC-GBXIJSLDSA-N	RM0135900
Tryptophan (Trp-4_1H)	Tryptophan	Organic acids	Amino acids and peptides	Amino acids	6305	16828	HMDB0000929		C00078	QIVBCDIJIAJPQS-VIFPVBQESA-N	RM0136024
Tyrosine (Tyr-3,5_1H)	Tyrosine	Organic acids	Amino acids and peptides	Amino acids	6057	17895	HMDB0000158		C00082	OUYCCCASQSFEME-QMMMGPOBSA-N	RM0135896
UDP-N-acetylgalactosamine (UDP-GalNAc_1H)	UDP-N-acetylgalactosamine	Nucleic acids	Pyrimidines	Pyrimidine rNDP	6419712	16650	HMDB0000304		C00203	LFTYTUAZOPRMMI-LDDHHVEYSA-N	RM0160954
UDP-glucose (UDP-Glc_1H)	UDP-glucose	Nucleic acids	Pyrimidines	Pyrimidine rNDP	53477679	167608	HMDB0000286		C00029	HSCJRCZFDFQWRP-LPTOLDDLSA-N	RM0135928
UXP (UXP-1'_1H)	UXP	Nucleic acids	Pyrimidines	Pyrimidine rNXP
UXP (UXP-1' 13C-A_1H)	UXP	Nucleic acids	Pyrimidines	Pyrimidine rNXP
UXP (UXP-5'_1H)	UXP	Nucleic acids	Pyrimidines	Pyrimidine rNXP
Valine (Val-B_1H)	Valine	Organic acids	Amino acids and peptides	Amino acids	6287	16414	HMDB0000883		C00183	KZSNJWFQEVHDMF-BYPYZUCNSA-N	RM0136018
Glucose (aGlc-1_1D 1H-13C-HSQC)	Glucose	Carbohydrates	Monosaccharides	Hexoses	5793	4167	HMDB0304632		C00031	WQZGKKKJIJFFOK-GASJEMHNSA-N	RM0135901
Alanine (Ala-3_1D 1H-13C-HSQC)	Alanine	Organic acids	Amino acids and peptides	Amino acids	5950	16977	HMDB0000161		C00041	QNAYBMKLOCPYGJ-REOHCLBHSA-N	RM0135898
AMP (AMP-3'_1D 1H-13C-HSQC)	AMP	Nucleic acids	Purines	Purine rNMP	6083	16027	HMDB0000045		C00020	UDMBCSSLTHHNCD-KQYNXXCUSA-N	RM0138921
ATP (ATP-4'_1D 1H-13C-HSQC)	ATP	Nucleic acids	Purines	Purine rNTP	5957	15422	HMDB0000538		C00002	ZKHQWZAMYRWXGA-KQYNXXCUSA-N	RM0138930
AXP (AXP-1'_1D 1H-13C-HSQC)	AXP	Nucleic acids	Purines	Purine rNXP
Glutamic acid (Glu-3a_1D 1H-13C-HSQC)	Glutamic acid	Organic acids	Amino acids and peptides	Amino acids	33032	16015	HMDB0000148		C00025	WHUUTDBJXJRKMK-VKHMYHEASA-N	RM0135893
Glutamic acid (Glu-4_1D 1H-13C-HSQC)	Glutamic acid	Organic acids	Amino acids and peptides	Amino acids	33032	16015	HMDB0000148		C00025	WHUUTDBJXJRKMK-VKHMYHEASA-N	RM0135893
Glutathione (GSH-4_1D 1H-13C-HSQC)	Glutathione	Organic acids	Amino acids and peptides	Tripeptides	124886	16856	HMDB0000125		C00051	RWSXRVCMGQZWBV-WDSKDSINSA-N	RM0135889
GTP (GTP_1D 1H-13C-HSQC)	GTP	Nucleic acids	Purines	Purine rNTP	135398633	15996	HMDB0001273		C00044	XKMLYUALXHKNFT-UUOKFMHZSA-N	RM0136084
Lactic acid (Lac-2_1D 1H-13C-HSQC)	Lactic acid	Organic acids	Short-chain acids	Short-chain acids	107689	422	HMDB0000190		C00186	JVTAAEKCZFNVCJ-REOHCLBHSA-N	RM0135904
Lactic acid/Threonine (Lac-3/Thr_1D 1H-13C-HSQC)	Lactic acid/Threonine	Organic acids
NAD (NAD+-N1'_1D 1H-13C-HSQC)	NAD	Nucleic acids	Nicotinamides	Nicotinamide dinucleotides	5892	44215	HMDB0000902		C00003	BAWFJGJZGIEFAR-NNYOXOHSSA-N	RM0133217
UXP (UXP-1'_1D 1H-13C-HSQC)	UXP	Nucleic acids	Pyrimidines	Pyrimidine rNXP
METABOLITES_END
#END