#METABOLOMICS WORKBENCH hormel101_20161021_131456_mwtab.txt DATATRACK_ID:762
VERSION             	1
CREATED_ON             	November 16, 2016, 2:50 pm
#PROJECT
PR:PROJECT_TITLE                 	Metabolic Dissection of Aberrant Glutamine Metabolism in Prostate Cancer
PR:PROJECT_TYPE                  	MS targeted analysis
PR:PROJECT_SUMMARY               	In this project, we propose to define the specific role(s) of three crucial
PR:PROJECT_SUMMARY               	oncogenic signaling cascades (androgen receptor, Myc and AMPK) in regulating
PR:PROJECT_SUMMARY               	glutamine metabolism in prostate cancer using 13C-labeled glutamine with mass
PR:PROJECT_SUMMARY               	spectrometry-based metabolic flux analyses.
PR:INSTITUTE                     	Mayo Clinic
PR:LABORATORY                    	Mayo Clinic Metabolomics Resource Core
PR:LAST_NAME                     	Frigo
PR:FIRST_NAME                    	Daniel
PR:ADDRESS                       	Department of Biology and Biochemistry, University of Houston, Houston, Texas
PR:ADDRESS                       	77204-5001
PR:EMAIL                         	frigo@uh.edu
PR:PHONE                         	832-842-8824
#STUDY
ST:STUDY_TITLE                   	Timecourse of palmitate isotopomers in LNCaP Cells
ST:STUDY_SUMMARY                 	Timecourse of palmitate isotopomers in LNCaP Cell pellets
ST:INSTITUTE                     	Mayo Clinic
ST:LABORATORY                    	Mayo Clinic Metabolomics Resource Core
ST:LAST_NAME                     	Frigo
ST:FIRST_NAME                    	Daniel
ST:ADDRESS                       	Department of Biology and Biochemistry, University of Houston, Houston, Texas
ST:ADDRESS                       	77204-5001
ST:EMAIL                         	frigo@uh.edu
ST:PHONE                         	832-842-8824
#SUBJECT
SU:SUBJECT_TYPE                  	Human
SU:SUBJECT_SPECIES               	Homo sapiens
SU:TAXONOMY_ID                   	9606
#SUBJECT_SAMPLE_FACTORS:         	SUBJECT(optional)[tab]SAMPLE[tab]FACTORS(NAME:VALUE pairs separated by |)[tab]Additional sample data
SUBJECT_SAMPLE_FACTORS           	DF-1	ms5457-1	Treatment:treated-vehicle (ethanol) | 13C media incubation time:24 hours	Amount=3 million cells; 1x PBS=50; assay=palmitate isotopomers
SUBJECT_SAMPLE_FACTORS           	DF-2	ms5457-2	Treatment:treated-100 pM R1881 | 13C media incubation time:24 hours	Amount=3 million cells; 1x PBS=50; assay=palmitate isotopomers
SUBJECT_SAMPLE_FACTORS           	DF-3	ms5457-3	Treatment:treated-1 nM R1881 | 13C media incubation time:24 hours	Amount=3 million cells; 1x PBS=50; assay=palmitate isotopomers
SUBJECT_SAMPLE_FACTORS           	DF-4	ms5457-4	Treatment:treated-10 nM R1881 | 13C media incubation time:24 hours	Amount=3 million cells; 1x PBS=50; assay=palmitate isotopomers
SUBJECT_SAMPLE_FACTORS           	DF-8	ms5457-5	Treatment:treated-vehicle (ethanol) | 13C media incubation time:72 hours	Amount=3 million cells; 1x PBS=50; assay=palmitate isotopomers
SUBJECT_SAMPLE_FACTORS           	DF-9	ms5457-6	Treatment:treated-100 pM R1881 | 13C media incubation time:72 hours	Amount=3 million cells; 1x PBS=50; assay=palmitate isotopomers
SUBJECT_SAMPLE_FACTORS           	DF-10	ms5457-7	Treatment:treated-1 nM R1881 | 13C media incubation time:72 hours	Amount=3 million cells; 1x PBS=50; assay=palmitate isotopomers
SUBJECT_SAMPLE_FACTORS           	DF-11	ms5457-8	Treatment:treated-10 nM R1881 | 13C media incubation time:72 hours	Amount=3 million cells; 1x PBS=50; assay=palmitate isotopomers
SUBJECT_SAMPLE_FACTORS           	DF-1	ms5457-9	Treatment:treated-vehicle (ethanol) | 13C media incubation time:24 hours	Amount=10 mls spent media; 1x PBS=-; assay=palmitate isotopomers
SUBJECT_SAMPLE_FACTORS           	DF-2	ms5457-10	Treatment:treated-100 pM R1881 | 13C media incubation time:24 hours	Amount=10 mls spent media; 1x PBS=-; assay=palmitate isotopomers
SUBJECT_SAMPLE_FACTORS           	DF-3	ms5457-11	Treatment:treated-1 nM R1881 | 13C media incubation time:24 hours	Amount=10 mls spent media; 1x PBS=-; assay=palmitate isotopomers
SUBJECT_SAMPLE_FACTORS           	DF-4	ms5457-12	Treatment:treated-10 nM R1881 | 13C media incubation time:24 hours	Amount=10 mls spent media; 1x PBS=-; assay=palmitate isotopomers
SUBJECT_SAMPLE_FACTORS           	DF-8	ms5457-13	Treatment:treated-vehicle (ethanol) | 13C media incubation time:72 hours	Amount=10 mls spent media; 1x PBS=-; assay=palmitate isotopomers
SUBJECT_SAMPLE_FACTORS           	DF-9	ms5457-14	Treatment:treated-100 pM R1881 | 13C media incubation time:72 hours	Amount=10 mls spent media; 1x PBS=-; assay=palmitate isotopomers
SUBJECT_SAMPLE_FACTORS           	DF-10	ms5457-15	Treatment:treated-1 nM R1881 | 13C media incubation time:72 hours	Amount=10 mls spent media; 1x PBS=-; assay=palmitate isotopomers
SUBJECT_SAMPLE_FACTORS           	DF-11	ms5457-16	Treatment:treated-10 nM R1881 | 13C media incubation time:72 hours	Amount=10 mls spent media; 1x PBS=-; assay=palmitate isotopomers
#COLLECTION
CO:COLLECTION_SUMMARY            	LNCaP cell model
#TREATMENT
TR:TREATMENT_SUMMARY             	LNCaP cells that were serum starved and incubated with 2mM [U-13C]glutamine and
TR:TREATMENT_SUMMARY             	treated with dose response of a synthetic androgen (R1881) for 24 or 72 hours.
#SAMPLEPREP
SP:SAMPLEPREP_SUMMARY            	Add 50 uL 1x PBS solution to each vial containing cells. Cap, vortex, sonicate 2
SP:SAMPLEPREP_SUMMARY            	x 20 sec bursts with vortexing inbetween and after Transfer entire contents of
SP:SAMPLEPREP_SUMMARY            	each vial into a labeled 1 dram vial. use 50 uL media for samples 9-16 use 50 uL
SP:SAMPLEPREP_SUMMARY            	plasma for QC Add 1.5 mL CHM solution to vial Add 20 uL 1:20 phenol red
SP:SAMPLEPREP_SUMMARY            	indicator Cap and vortex on multivortexer for 10 minutes Centrifuge @ 4C @ 3000
SP:SAMPLEPREP_SUMMARY            	rpm for 10 minutes Aspirate and discard top phase hard to see the phase
SP:SAMPLEPREP_SUMMARY            	separation due to the light color (yellow). Dry bottom phase in speed vac
SP:SAMPLEPREP_SUMMARY            	Reconstitute samples and standards with 25 uL buffer A. (do triplicate
SP:SAMPLEPREP_SUMMARY            	injections)
#CHROMATOGRAPHY
CH:CHROMATOGRAPHY_TYPE           	Reversed phase
CH:INSTRUMENT_NAME               	Agilent 1290 Infinity
CH:COLUMN_NAME                   	Waters Acquity BEH C18 (150 x 2.1mm, 1.7um)
#ANALYSIS
AN:ANALYSIS_TYPE                 	MS
#MS
MS:MS_COMMENTS                   	-
MS:INSTRUMENT_NAME               	Agilent 6460 QQQ
MS:INSTRUMENT_TYPE               	Triple quadrupole
MS:MS_TYPE                       	ESI
MS:ION_MODE                      	UNSPECIFIED
#MS_METABOLITE_DATA
MS_METABOLITE_DATA:UNITS         	13C-palmitate % enrichment
MS_METABOLITE_DATA_START
Samples	ms5457-1	ms5457-2	ms5457-3	ms5457-4	ms5457-5	ms5457-6	ms5457-7	ms5457-8	ms5457-9	ms5457-10	ms5457-11	ms5457-12	ms5457-13	ms5457-14	ms5457-15	ms5457-16
Factors	Treatment:treated-vehicle (ethanol) | 13C media incubation time:24 hours	Treatment:treated-100 pM R1881 | 13C media incubation time:24 hours	Treatment:treated-1 nM R1881 | 13C media incubation time:24 hours	Treatment:treated-10 nM R1881 | 13C media incubation time:24 hours	Treatment:treated-vehicle (ethanol) | 13C media incubation time:72 hours	Treatment:treated-100 pM R1881 | 13C media incubation time:72 hours	Treatment:treated-1 nM R1881 | 13C media incubation time:72 hours	Treatment:treated-10 nM R1881 | 13C media incubation time:72 hours	Treatment:treated-vehicle (ethanol) | 13C media incubation time:24 hours	Treatment:treated-100 pM R1881 | 13C media incubation time:24 hours	Treatment:treated-1 nM R1881 | 13C media incubation time:24 hours	Treatment:treated-10 nM R1881 | 13C media incubation time:24 hours	Treatment:treated-vehicle (ethanol) | 13C media incubation time:72 hours	Treatment:treated-100 pM R1881 | 13C media incubation time:72 hours	Treatment:treated-1 nM R1881 | 13C media incubation time:72 hours	Treatment:treated-10 nM R1881 | 13C media incubation time:72 hours
m0 palmitate isotopomers	88.99	87.46	87.81	83.06	83.13	77.56	78.33	76.18	99.97	99.83	99.82	99.91	99.69	99.67	99.86	99.99
m1 palmitate isotopomers	0.28	0.65	0.67	1.11	1.67	2.29	2.41	2.31	0.00	0.17	0.16	0.00	0.31	0.33	0.13	0.00
m2 palmitate isotopomers	5.91	5.95	6.14	8.03	9.81	13.05	12.92	14.00	0.03	0.00	0.00	0.08	0.00	0.00	0.00	0.00
m3 palmitate isotopomers	0.31	0.41	0.37	0.56	0.71	0.92	0.68	0.82	0.00	0.00	0.01	0.00	0.00	0.01	0.01	0.01
m4 palmitate isotopomers	2.92	3.44	3.19	4.55	3.34	4.42	4.16	4.76	0.00	0.00	0.00	0.00	0.00	0.00	0.00	0.00
m5 palmitate isotopomers	0.08	0.13	0.09	0.15	0.12	0.17	0.10	0.18	0.00	0.00	0.00	0.00	0.00	0.00	0.00	0.00
m6 palmitate isotopomers	1.06	1.36	1.18	1.78	0.90	1.21	1.05	1.33	0.00	0.00	0.00	0.00	0.00	0.00	0.00	0.00
m7 palmitate isotopomers	0.00	0.00	0.00	0.00	0.00	0.01	0.02	0.01	0.00	0.00	0.00	0.00	0.00	0.00	0.00	0.00
m8 palmitate isotopomers	0.35	0.45	0.44	0.58	0.24	0.30	0.26	0.33	0.00	0.00	0.00	0.00	0.00	0.00	0.00	0.00
m9 palmitate isotopomers	0.00	0.00	0.00	0.00	0.00	0.00	0.00	0.00	0.00	0.00	0.00	0.00	0.00	0.00	0.00	0.00
m10 palmitate isotopomers	0.11	0.15	0.13	0.17	0.07	0.08	0.07	0.09	0.00	0.00	0.00	0.00	0.00	0.00	0.00	0.00
m11 palmitate isotopomers	0.00	0.00	0.00	0.00	0.00	0.00	0.00	0.00	0.00	0.00	0.00	0.00	0.00	0.00	0.00	0.00
m12 palmitate isotopomers	0.00	0.00	0.00	0.00	0.00	0.00	0.00	0.00	0.00	0.00	0.00	0.00	0.00	0.00	0.00	0.00
m13 palmitate isotopomers	0.00	0.00	0.00	0.00	0.00	0.00	0.00	0.00	0.00	0.00	0.00	0.00	0.00	0.00	0.00	0.00
m14 palmitate isotopomers	0.00	0.00	0.00	0.00	0.00	0.00	0.00	0.00	0.00	0.00	0.00	0.00	0.00	0.00	0.00	0.00
m15 palmitate isotopomers	0.00	0.00	0.00	0.00	0.00	0.00	0.00	0.00	0.00	0.00	0.00	0.00	0.00	0.00	0.00	0.00
m16 palmitate isotopomers	0.00	0.00	0.00	0.00	0.00	0.00	0.00	0.00	0.00	0.00	0.00	0.00	0.00	0.00	0.00	0.00
MS_METABOLITE_DATA_END
#METABOLITES
METABOLITES_START
metabolite_name
m0 palmitate isotopomers
m1 palmitate isotopomers
m2 palmitate isotopomers
m3 palmitate isotopomers
m4 palmitate isotopomers
m5 palmitate isotopomers
m6 palmitate isotopomers
m7 palmitate isotopomers
m8 palmitate isotopomers
m9 palmitate isotopomers
m10 palmitate isotopomers
m11 palmitate isotopomers
m12 palmitate isotopomers
m13 palmitate isotopomers
m14 palmitate isotopomers
m15 palmitate isotopomers
m16 palmitate isotopomers
METABOLITES_END
#END