#METABOLOMICS WORKBENCH mjosephkeller_20230926_123700 DATATRACK_ID:4336 STUDY_ID:ST002890 ANALYSIS_ID:AN004748 PROJECT_ID:PR001803
VERSION             	1
CREATED_ON             	September 29, 2023, 6:40 pm
#PROJECT
PR:PROJECT_TITLE                 	Characterization of the in vivo deuteration of native phospholipids by mass
PR:PROJECT_TITLE                 	spectrometry yields guidelines for their regiospecific customization
PR:PROJECT_SUMMARY               	Customization of deuterated biomolecules is vital for many advanced biological
PR:PROJECT_SUMMARY               	experiments, including neutron scattering. However, because it is challenging to
PR:PROJECT_SUMMARY               	control the proportion and regiospecificity of deuterium incorporation in live
PR:PROJECT_SUMMARY               	systems, often only two or three synthetic lipids are mixed together to form
PR:PROJECT_SUMMARY               	simplistic model membranes. This limits the applicability and biological
PR:PROJECT_SUMMARY               	accuracy of the results generated with these synthetic membranes. Despite some
PR:PROJECT_SUMMARY               	limited prior examination of deuterating E. coli lipids in vivo, this approach
PR:PROJECT_SUMMARY               	has not been widely implemented. Here, an extensive mass spectrometry-based
PR:PROJECT_SUMMARY               	profiling of E. coli phospholipid deuteration states with several different
PR:PROJECT_SUMMARY               	growth media was performed and a computational method to describe deuterium
PR:PROJECT_SUMMARY               	distributions with a one-number summary is introduced. The deuteration states of
PR:PROJECT_SUMMARY               	thirty-six lipid species were quantitatively profiled in fifteen different
PR:PROJECT_SUMMARY               	growth conditions and tandem mass spectrometry was used to reveal deuterium
PR:PROJECT_SUMMARY               	localization. Regressions were employed to enable the prediction of lipid
PR:PROJECT_SUMMARY               	deuteration for untested conditions. Small-angle neutron scattering was
PR:PROJECT_SUMMARY               	performed on select deuterated lipid samples, which validated the deuteration
PR:PROJECT_SUMMARY               	states calculated from the mass spectral data. Based on these experiments,
PR:PROJECT_SUMMARY               	guidelines for the design of specifically deuterated phospholipids are
PR:PROJECT_SUMMARY               	described. This unlocks even greater capabilities from neutron-based techniques,
PR:PROJECT_SUMMARY               	enabling experiments that were formerly impossible.
PR:INSTITUTE                     	University of Tennessee
PR:DEPARTMENT                    	Genome Science and Technology (Bredesen Center)
PR:LAST_NAME                     	Matthew
PR:FIRST_NAME                    	Keller
PR:ADDRESS                       	The Bredesen Center for Interdisciplinary Research and Graduate Education 444
PR:ADDRESS                       	Greve Hall, 821 Volunteer Blvd
PR:EMAIL                         	qrh579@vols.utk.edu
PR:PHONE                         	18659747999
PR:PUBLICATIONS                  	In review at Analytical Chemistry
PR:CONTRIBUTORS                  	Qiu Zhang, Shuo Qian, Brian Sanders, Hugh O'Neill, Robert Hettich
#STUDY
ST:STUDY_TITLE                   	Characterization of the in vivo deuteration of native phospholipids by mass
ST:STUDY_TITLE                   	spectrometry yields guidelines for their regiospecific customization
ST:STUDY_SUMMARY                 	Customization of deuterated biomolecules is vital for many advanced biological
ST:STUDY_SUMMARY                 	experiments, including neutron scattering. However, because it is challenging to
ST:STUDY_SUMMARY                 	control the proportion and regiospecificity of deuterium incorporation in live
ST:STUDY_SUMMARY                 	systems, often only two or three synthetic lipids are mixed together to form
ST:STUDY_SUMMARY                 	simplistic model membranes. This limits the applicability and biological
ST:STUDY_SUMMARY                 	accuracy of the results generated with these synthetic membranes. Despite some
ST:STUDY_SUMMARY                 	limited prior examination of deuterating E. coli lipids in vivo, this approach
ST:STUDY_SUMMARY                 	has not been widely implemented. Here, an extensive mass spectrometry-based
ST:STUDY_SUMMARY                 	profiling of E. coli phospholipid deuteration states with several different
ST:STUDY_SUMMARY                 	growth media was performed and a computational method to describe deuterium
ST:STUDY_SUMMARY                 	distributions with a one-number summary is introduced. The deuteration states of
ST:STUDY_SUMMARY                 	thirty-six lipid species were quantitatively profiled in fifteen different
ST:STUDY_SUMMARY                 	growth conditions and tandem mass spectrometry was used to reveal deuterium
ST:STUDY_SUMMARY                 	localization. Regressions were employed to enable the prediction of lipid
ST:STUDY_SUMMARY                 	deuteration for untested conditions. Small-angle neutron scattering was
ST:STUDY_SUMMARY                 	performed on select deuterated lipid samples, which validated the deuteration
ST:STUDY_SUMMARY                 	states calculated from the mass spectral data. Based on these experiments,
ST:STUDY_SUMMARY                 	guidelines for the design of specifically deuterated phospholipids are
ST:STUDY_SUMMARY                 	described. This unlocks even greater capabilities from neutron-based techniques,
ST:STUDY_SUMMARY                 	enabling experiments that were formerly impossible.
ST:INSTITUTE                     	University of Tennessee
ST:DEPARTMENT                    	Genome Science and Technology (Bredesen Center)
ST:LAST_NAME                     	Matthew
ST:FIRST_NAME                    	Keller
ST:ADDRESS                       	The Bredesen Center for Interdisciplinary Research and Graduate Education 444
ST:ADDRESS                       	Greve Hall, 821 Volunteer Blvd. Knoxville, TN 37996-3394
ST:EMAIL                         	qrh579@vols.utk.edu
ST:PHONE                         	18659747999
#SUBJECT
SU:SUBJECT_TYPE                  	Bacteria
SU:SUBJECT_SPECIES               	Escherichia coli
SU:GENOTYPE_STRAIN               	BL21(DE3)
#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           	-	50D-Gly/0_1-neg	Carbon source:H-Glucose | Percent D2O:100 | MS ionization mode:negative	Technical replicate=1; RAW_FILE_NAME=50D-Gly_0_1-neg.mzML
SUBJECT_SAMPLE_FACTORS           	-	50D-Gly/0_2-neg	Carbon source:H-Glucose | Percent D2O:100 | MS ionization mode:negative	Technical replicate=1; RAW_FILE_NAME=50D-Gly_0_2-neg.mzML
SUBJECT_SAMPLE_FACTORS           	-	50D-Gly/0_3-neg	Carbon source:H-Glucose | Percent D2O:100 | MS ionization mode:negative	Technical replicate=1; RAW_FILE_NAME=50D-Gly_0_3-neg.mzML
SUBJECT_SAMPLE_FACTORS           	-	50D-Gly/100_1-neg	Carbon source:D-Glucose | Percent D2O:100 | MS ionization mode:negative	Technical replicate=1; RAW_FILE_NAME=50D-Gly_100_1-neg.mzML
SUBJECT_SAMPLE_FACTORS           	-	50D-Gly/100_2-neg	Carbon source:D-Glucose | Percent D2O:100 | MS ionization mode:negative	Technical replicate=1; RAW_FILE_NAME=50D-Gly_100_2-neg.mzML
SUBJECT_SAMPLE_FACTORS           	-	50D-Gly/100_3-neg	Carbon source:D-Glucose | Percent D2O:100 | MS ionization mode:negative	Technical replicate=1; RAW_FILE_NAME=50D-Gly_100_3-neg.mzML
SUBJECT_SAMPLE_FACTORS           	-	D-Glu/0_1-neg	Carbon source:H-Glucose | Percent D2O:25 | MS ionization mode:negative	Technical replicate=1; RAW_FILE_NAME=D-Glu_0_1_neg.mzML
SUBJECT_SAMPLE_FACTORS           	-	D-Glu/0_2-neg	Carbon source:H-Glucose | Percent D2O:25 | MS ionization mode:negative	Technical replicate=1; RAW_FILE_NAME=D-Glu_0_2_neg.mzML
SUBJECT_SAMPLE_FACTORS           	-	D-Glu/0_3-neg	Carbon source:H-Glucose | Percent D2O:25 | MS ionization mode:negative	Technical replicate=1; RAW_FILE_NAME=D-Glu_0_3_neg.mzML
SUBJECT_SAMPLE_FACTORS           	-	D-Glu/100-neg_tech-rep1	Carbon source:D-Glucose | Percent D2O:0 | MS ionization mode:negative	Technical replicate=1; RAW_FILE_NAME=D-Glu_100_tech1-neg.mzML
SUBJECT_SAMPLE_FACTORS           	-	D-Glu/100-neg_tech-rep2	Carbon source:D-Glucose | Percent D2O:0 | MS ionization mode:negative	Technical replicate=2; RAW_FILE_NAME=D-Glu_100_tech2-neg.mzML
SUBJECT_SAMPLE_FACTORS           	-	D-Gly/0_1-neg	Carbon source:H-Glucose | Percent D2O:50 | MS ionization mode:negative	Technical replicate=1; RAW_FILE_NAME=D-Gly_0_1_neg.mzML
SUBJECT_SAMPLE_FACTORS           	-	D-Gly/0_2-neg	Carbon source:H-Glucose | Percent D2O:50 | MS ionization mode:negative	Technical replicate=1; RAW_FILE_NAME=D-Gly_0_2_neg.mzML
SUBJECT_SAMPLE_FACTORS           	-	D-Gly/0_3-neg	Carbon source:H-Glucose | Percent D2O:50 | MS ionization mode:negative	Technical replicate=1; RAW_FILE_NAME=D-Gly_0_3_neg.mzML
SUBJECT_SAMPLE_FACTORS           	-	D-Gly/100_1-neg	Carbon source:H-Glucose | Percent D2O:75 | MS ionization mode:negative	Technical replicate=1; RAW_FILE_NAME=D-Gly_100_1_neg.mzML
SUBJECT_SAMPLE_FACTORS           	-	D-Gly/100_2-neg	Carbon source:H-Glucose | Percent D2O:75 | MS ionization mode:negative	Technical replicate=1; RAW_FILE_NAME=D-Gly_100_2_neg.mzML
SUBJECT_SAMPLE_FACTORS           	-	D-Gly/100_3-neg	Carbon source:H-Glucose | Percent D2O:75 | MS ionization mode:negative	Technical replicate=1; RAW_FILE_NAME=D-Gly_100_3_neg.mzML
SUBJECT_SAMPLE_FACTORS           	-	H-Glu/100-neg_tech-rep1	Carbon source:D-Glycerol | Percent D2O:100 | MS ionization mode:negative	Technical replicate=1; RAW_FILE_NAME=H-Glu_100_tech1-neg.mzML
SUBJECT_SAMPLE_FACTORS           	-	H-Glu/100-neg_tech-rep2	Carbon source:D-Glycerol | Percent D2O:100 | MS ionization mode:negative	Technical replicate=2; RAW_FILE_NAME=H-Glu_100_tech2-neg.mzML
SUBJECT_SAMPLE_FACTORS           	-	H-Glu/25_1-neg	Carbon source:50/50 H/D-Glycerol | Percent D2O:0 | MS ionization mode:negative	Technical replicate=1; RAW_FILE_NAME=H-Glu_25_1-neg.mzML
SUBJECT_SAMPLE_FACTORS           	-	H-Glu/25_2-neg	Carbon source:50/50 H/D-Glycerol | Percent D2O:0 | MS ionization mode:negative	Technical replicate=1; RAW_FILE_NAME=H-Glu_25_2-neg.mzML
SUBJECT_SAMPLE_FACTORS           	-	H-Glu/25_3-neg	Carbon source:50/50 H/D-Glycerol | Percent D2O:0 | MS ionization mode:negative	Technical replicate=1; RAW_FILE_NAME=H-Glu_25_3-neg.mzML
SUBJECT_SAMPLE_FACTORS           	-	H-Glu/50_1-neg	Carbon source:D-Glycerol | Percent D2O:0 | MS ionization mode:negative	Technical replicate=1; RAW_FILE_NAME=H-Glu_50_1-neg.mzML
SUBJECT_SAMPLE_FACTORS           	-	H-Glu/50_2-neg	Carbon source:D-Glycerol | Percent D2O:0 | MS ionization mode:negative	Technical replicate=1; RAW_FILE_NAME=H-Glu_50_2-neg.mzML
SUBJECT_SAMPLE_FACTORS           	-	H-Glu/50_3-neg	Carbon source:D-Glycerol | Percent D2O:0 | MS ionization mode:negative	Technical replicate=1; RAW_FILE_NAME=H-Glu_50_3-neg.mzML
SUBJECT_SAMPLE_FACTORS           	-	H-Glu/75_1-neg	Carbon source:50/50 H/D-Glycerol | Percent D2O:100 | MS ionization mode:negative	Technical replicate=1; RAW_FILE_NAME=H-Glu_75_1-neg.mzML
SUBJECT_SAMPLE_FACTORS           	-	H-Glu/75_2-neg	Carbon source:50/50 H/D-Glycerol | Percent D2O:100 | MS ionization mode:negative	Technical replicate=1; RAW_FILE_NAME=H-Glu_75_2-neg.mzML
SUBJECT_SAMPLE_FACTORS           	-	H-Glu/75_3-neg	Carbon source:50/50 H/D-Glycerol | Percent D2O:100 | MS ionization mode:negative	Technical replicate=1; RAW_FILE_NAME=H-Glu_75_3-neg.mzML
SUBJECT_SAMPLE_FACTORS           	-	H-Glu/75_repeat_1-neg	Carbon source:50/50 H/D-Glycerol | Percent D2O:100 | MS ionization mode:negative	Technical replicate=1; RAW_FILE_NAME=H-Glu_75_1-neg_2ndbatch.mzML
SUBJECT_SAMPLE_FACTORS           	-	H-Glu/75_repeat_2-neg	Carbon source:50/50 H/D-Glycerol | Percent D2O:100 | MS ionization mode:negative	Technical replicate=1; RAW_FILE_NAME=H-Glu_75_2-neg_2ndbatch.mzML
SUBJECT_SAMPLE_FACTORS           	-	H-Glu/75_repeat_3-neg	Carbon source:50/50 H/D-Glycerol | Percent D2O:100 | MS ionization mode:negative	Technical replicate=1; RAW_FILE_NAME=H-Glu_75_3-neg_2ndbatch.mzML
SUBJECT_SAMPLE_FACTORS           	-	H-Gly/0-neg_tech-rep1	Carbon source:H-Glycerol | Percent D2O:0 | MS ionization mode:negative	Technical replicate=1; RAW_FILE_NAME=H-Gly_0_tech1-neg.mzML
SUBJECT_SAMPLE_FACTORS           	-	H-Gly/0-neg_tech-rep2	Carbon source:H-Glycerol | Percent D2O:0 | MS ionization mode:negative	Technical replicate=2; RAW_FILE_NAME=H-Gly_0_tech2-neg.mzML
SUBJECT_SAMPLE_FACTORS           	-	H-Gly/100_1-neg	Carbon source:H-Glycerol | Percent D2O:100 | MS ionization mode:negative	Technical replicate=1; RAW_FILE_NAME=H-Gly_100_1-neg.mzML
SUBJECT_SAMPLE_FACTORS           	-	H-Gly/100_2-neg	Carbon source:H-Glycerol | Percent D2O:100 | MS ionization mode:negative	Technical replicate=1; RAW_FILE_NAME=H-Gly_100_2-neg.mzML
SUBJECT_SAMPLE_FACTORS           	-	H-Gly/100_3-neg	Carbon source:H-Glycerol | Percent D2O:100 | MS ionization mode:negative	Technical replicate=1; RAW_FILE_NAME=H-Gly_100_3-neg.mzML
SUBJECT_SAMPLE_FACTORS           	-	H-Gly/25-neg_tech-rep1	Carbon source:H-Glycerol | Percent D2O:25 | MS ionization mode:negative	Technical replicate=1; RAW_FILE_NAME=H-Gly_25_tech1-neg.mzML
SUBJECT_SAMPLE_FACTORS           	-	H-Gly/25-neg_tech-rep2	Carbon source:H-Glycerol | Percent D2O:25 | MS ionization mode:negative	Technical replicate=2; RAW_FILE_NAME=H-Gly_25_tech2-neg.mzML
SUBJECT_SAMPLE_FACTORS           	-	H-Gly/50-neg_tech-rep1	Carbon source:H-Glycerol | Percent D2O:50 | MS ionization mode:negative	Technical replicate=1; RAW_FILE_NAME=H-Gly_50_tech1-neg.mzML
SUBJECT_SAMPLE_FACTORS           	-	H-Gly/50-neg_tech-rep2	Carbon source:H-Glycerol | Percent D2O:50 | MS ionization mode:negative	Technical replicate=2; RAW_FILE_NAME=H-Gly_50_tech2-neg.mzML
SUBJECT_SAMPLE_FACTORS           	-	H-Gly/75-neg_tech-rep1	Carbon source:H-Glycerol | Percent D2O:75 | MS ionization mode:negative	Technical replicate=1; RAW_FILE_NAME=H-Gly_75_tech1-neg.mzML
SUBJECT_SAMPLE_FACTORS           	-	H-Gly/75-neg_tech-rep2	Carbon source:H-Glycerol | Percent D2O:75 | MS ionization mode:negative	Technical replicate=2; RAW_FILE_NAME=H-Gly_75_tech2-neg.mzML
SUBJECT_SAMPLE_FACTORS           	-	50D-Gly/0_1-pos	Carbon source:H-Glucose | Percent D2O:100 | MS ionization mode:positive	Technical replicate=1; RAW_FILE_NAME=50D-Gly_0_1-pos.mzML
SUBJECT_SAMPLE_FACTORS           	-	50D-Gly/0_2-pos	Carbon source:H-Glucose | Percent D2O:100 | MS ionization mode:positive	Technical replicate=1; RAW_FILE_NAME=50D-Gly_0_2-pos.mzML
SUBJECT_SAMPLE_FACTORS           	-	50D-Gly/0_3-pos	Carbon source:H-Glucose | Percent D2O:100 | MS ionization mode:positive	Technical replicate=1; RAW_FILE_NAME=50D-Gly_0_3-pos.mzML
SUBJECT_SAMPLE_FACTORS           	-	50D-Gly/100_1-pos	Carbon source:D-Glucose | Percent D2O:100 | MS ionization mode:positive	Technical replicate=1; RAW_FILE_NAME=50D-Gly_100_1-pos.mzML
SUBJECT_SAMPLE_FACTORS           	-	50D-Gly/100_2-pos	Carbon source:D-Glucose | Percent D2O:100 | MS ionization mode:positive	Technical replicate=1; RAW_FILE_NAME=50D-Gly_100_2-pos.mzML
SUBJECT_SAMPLE_FACTORS           	-	50D-Gly/100_3-pos	Carbon source:D-Glucose | Percent D2O:100 | MS ionization mode:positive	Technical replicate=1; RAW_FILE_NAME=50D-Gly_100_3-pos.mzML
SUBJECT_SAMPLE_FACTORS           	-	D-Glu/0_1-pos	Carbon source:H-Glucose | Percent D2O:25 | MS ionization mode:positive	Technical replicate=1; RAW_FILE_NAME=D-Glu_0_1_pos.mzML
SUBJECT_SAMPLE_FACTORS           	-	D-Glu/0_2-pos	Carbon source:H-Glucose | Percent D2O:25 | MS ionization mode:positive	Technical replicate=1; RAW_FILE_NAME=D-Glu_0_2_pos.mzML
SUBJECT_SAMPLE_FACTORS           	-	D-Glu/0_3-pos	Carbon source:H-Glucose | Percent D2O:25 | MS ionization mode:positive	Technical replicate=1; RAW_FILE_NAME=D-Glu_0_3_pos.mzML
SUBJECT_SAMPLE_FACTORS           	-	D-Glu/100-pos	Carbon source:D-Glucose | Percent D2O:0 | MS ionization mode:positive	Technical replicate=1; RAW_FILE_NAME=D-Glu_100_pos.mzML
SUBJECT_SAMPLE_FACTORS           	-	D-Gly/0_1-pos	Carbon source:H-Glucose | Percent D2O:50 | MS ionization mode:positive	Technical replicate=1; RAW_FILE_NAME=D-Gly_0_1_pos.mzML
SUBJECT_SAMPLE_FACTORS           	-	D-Gly/0_2-pos	Carbon source:H-Glucose | Percent D2O:50 | MS ionization mode:positive	Technical replicate=1; RAW_FILE_NAME=D-Gly_0_2_pos.mzML
SUBJECT_SAMPLE_FACTORS           	-	D-Gly/0_3-pos	Carbon source:H-Glucose | Percent D2O:50 | MS ionization mode:positive	Technical replicate=1; RAW_FILE_NAME=D-Gly_0_3_pos.mzML
SUBJECT_SAMPLE_FACTORS           	-	D-Gly/100_1-pos	Carbon source:H-Glucose | Percent D2O:75 | MS ionization mode:positive	Technical replicate=1; RAW_FILE_NAME=D-Gly_100_1_pos.mzML
SUBJECT_SAMPLE_FACTORS           	-	D-Gly/100_2-pos	Carbon source:H-Glucose | Percent D2O:75 | MS ionization mode:positive	Technical replicate=1; RAW_FILE_NAME=D-Gly_100_2_pos.mzML
SUBJECT_SAMPLE_FACTORS           	-	D-Gly/100_3-pos	Carbon source:H-Glucose | Percent D2O:75 | MS ionization mode:positive	Technical replicate=1; RAW_FILE_NAME=D-Gly_100_3_pos.mzML
SUBJECT_SAMPLE_FACTORS           	-	H-Glu/100-pos	Carbon source:D-Glycerol | Percent D2O:100 | MS ionization mode:positive	Technical replicate=1; RAW_FILE_NAME=H-Glu_100_pos.mzML
SUBJECT_SAMPLE_FACTORS           	-	H-Glu/25_1-pos	Carbon source:50/50 H/D-Glycerol | Percent D2O:0 | MS ionization mode:positive	Technical replicate=1; RAW_FILE_NAME=H-Glu_25_1-pos.mzML
SUBJECT_SAMPLE_FACTORS           	-	H-Glu/25_2-pos	Carbon source:50/50 H/D-Glycerol | Percent D2O:0 | MS ionization mode:positive	Technical replicate=1; RAW_FILE_NAME=H-Glu_25_2-pos.mzML
SUBJECT_SAMPLE_FACTORS           	-	H-Glu/25_3-pos	Carbon source:50/50 H/D-Glycerol | Percent D2O:0 | MS ionization mode:positive	Technical replicate=1; RAW_FILE_NAME=H-Glu_25_3-pos.mzML
SUBJECT_SAMPLE_FACTORS           	-	H-Glu/50_1-pos	Carbon source:D-Glycerol | Percent D2O:0 | MS ionization mode:positive	Technical replicate=1; RAW_FILE_NAME=H-Glu_50_1-pos.mzML
SUBJECT_SAMPLE_FACTORS           	-	H-Glu/50_2-pos	Carbon source:D-Glycerol | Percent D2O:0 | MS ionization mode:positive	Technical replicate=1; RAW_FILE_NAME=H-Glu_50_2-pos.mzML
SUBJECT_SAMPLE_FACTORS           	-	H-Glu/50_3-pos	Carbon source:D-Glycerol | Percent D2O:0 | MS ionization mode:positive	Technical replicate=1; RAW_FILE_NAME=H-Glu_50_3-pos.mzML
SUBJECT_SAMPLE_FACTORS           	-	H-Glu/75_1-pos	Carbon source:50/50 H/D-Glycerol | Percent D2O:100 | MS ionization mode:positive	Technical replicate=1; RAW_FILE_NAME=H-Glu_75_1-pos.mzML
SUBJECT_SAMPLE_FACTORS           	-	H-Glu/75_2-pos	Carbon source:50/50 H/D-Glycerol | Percent D2O:100 | MS ionization mode:positive	Technical replicate=1; RAW_FILE_NAME=H-Glu_75_2-pos.mzML
SUBJECT_SAMPLE_FACTORS           	-	H-Glu/75_3-pos	Carbon source:50/50 H/D-Glycerol | Percent D2O:100 | MS ionization mode:positive	Technical replicate=1; RAW_FILE_NAME=H-Glu_75_3-pos.mzML
SUBJECT_SAMPLE_FACTORS           	-	H-Gly/0-pos	Carbon source:H-Glycerol | Percent D2O:0 | MS ionization mode:positive	Technical replicate=1; RAW_FILE_NAME=H-Gly_0_pos.mzML
SUBJECT_SAMPLE_FACTORS           	-	H-Gly/100_1-pos	Carbon source:H-Glycerol | Percent D2O:100 | MS ionization mode:positive	Technical replicate=1; RAW_FILE_NAME=H-Gly_100_1-pos.mzML
SUBJECT_SAMPLE_FACTORS           	-	H-Gly/100_2-pos	Carbon source:H-Glycerol | Percent D2O:100 | MS ionization mode:positive	Technical replicate=1; RAW_FILE_NAME=H-Gly_100_2-pos.mzML
SUBJECT_SAMPLE_FACTORS           	-	H-Gly/100_3-pos	Carbon source:H-Glycerol | Percent D2O:100 | MS ionization mode:positive	Technical replicate=1; RAW_FILE_NAME=H-Gly_100_3-pos.mzML
SUBJECT_SAMPLE_FACTORS           	-	H-Gly/25-pos	Carbon source:H-Glycerol | Percent D2O:25 | MS ionization mode:positive	Technical replicate=1; RAW_FILE_NAME=H-Gly_25_pos.mzML
SUBJECT_SAMPLE_FACTORS           	-	H-Gly/50-pos	Carbon source:H-Glycerol | Percent D2O:50 | MS ionization mode:positive	Technical replicate=1; RAW_FILE_NAME=H-Gly_50_pos.mzML
SUBJECT_SAMPLE_FACTORS           	-	H-Gly/75-pos	Carbon source:H-Glycerol | Percent D2O:75 | MS ionization mode:positive	Technical replicate=1; RAW_FILE_NAME=H-Gly_75_pos.mzML
#COLLECTION
CO:COLLECTION_SUMMARY            	Lipids were extracted from E coli cell pellets following the Matyash protocol
CO:COLLECTION_SUMMARY            	with slight variations described in the manuscript supplemental. Cell lysis was
CO:COLLECTION_SUMMARY            	performed with bead beating or ultrasonication. Extraction solvents were in the
CO:COLLECTION_SUMMARY            	ratio 10:3:25 MTBE:methanol:water. See publication for more detail.
CO:COLLECTION_PROTOCOL_FILENAME  	Deuterated_Lipids_Methods_Summary.pdf
CO:SAMPLE_TYPE                   	Bacterial cells
#TREATMENT
TR:TREATMENT_SUMMARY             	E. coli was grown in Enfors minimal media (deuterated or otherwise) with
TR:TREATMENT_SUMMARY             	different carbon sources including glycerol, glucose, D-glycerol, and D-glucose.
TR:CELL_MEDIA                    	Enfors minimal media
TR:CELL_HARVESTING               	30 mL culture collected by centrifugation at 4k x g
#SAMPLEPREP
SP:SAMPLEPREP_SUMMARY            	Lipids were extracted from E coli cell pellets following the Matyash protocol
SP:SAMPLEPREP_SUMMARY            	with slight variations described in the manuscript supplemental. Cell lysis was
SP:SAMPLEPREP_SUMMARY            	performed with bead beating or ultrasonication. Extraction solvents were in the
SP:SAMPLEPREP_SUMMARY            	ratio 10:3:25 MTBE:methanol:water. See publication for more detail.
SP:SAMPLEPREP_PROTOCOL_FILENAME  	Deuterated_Lipids_Methods_Summary.pdf
SP:EXTRACTION_METHOD             	Matyash (MTBE) varient
SP:SAMPLE_RESUSPENSION           	8:23:69 butanol:isopropanol:water
#CHROMATOGRAPHY
CH:CHROMATOGRAPHY_SUMMARY        	Because the columns were packed in-house, there is some variation in the total
CH:CHROMATOGRAPHY_SUMMARY        	length (see column description below).
CH:CHROMATOGRAPHY_TYPE           	Reversed phase
CH:INSTRUMENT_NAME               	Thermo Dionex Ultimate 3000 RS
CH:COLUMN_NAME                   	In-house packed nano column (ID 100 μm, Kinetex 1.7 μm C18 (100 Å)
CH:COLUMN_NAME                   	[Phenomenex], 12-15 cm.
CH:SOLVENT_A                     	60% acetonitrile/40% water; ~0.4g/L ammonium acetate
CH:SOLVENT_B                     	90% sopropanol/10% acetonitrile; ~0.4g/L ammonium acetate
CH:FLOW_GRADIENT                 	Solvent Gradient: Time (min) 0 1 2.5 7.5 13.5 48.5 58.5 75.5 77.5 87 Solvent A
CH:FLOW_GRADIENT                 	(%) 99 99 70 65 45 30 1 1 99 99 Solvent B (%) 1 1 30 45 55 70 99 99 1 1
CH:FLOW_RATE                     	Split flow nano-chromatography. The actual flow rate at the column is dependent
CH:FLOW_RATE                     	on a variety of factors and an accurate measurement of the flow rate is
CH:FLOW_RATE                     	impractical. The flow rate at the macro pumps were set at either 0.05 or 0.1
CH:FLOW_RATE                     	mL/min depending on the waste line's backpressure.
CH:COLUMN_TEMPERATURE            	room temp
CH:METHODS_FILENAME              	Deuterated_Lipids_Methods_Summary.pdf
#ANALYSIS
AN:ANALYSIS_TYPE                 	MS
AN:ANALYSIS_PROTOCOL_FILE        	Deuterated_Lipids_Methods_Summary.pdf
#MS
MS:INSTRUMENT_NAME               	Thermo Velos Pro Orbitrap
MS:INSTRUMENT_TYPE               	LTQ-FT
MS:MS_TYPE                       	ESI
MS:ION_MODE                      	NEGATIVE
MS:MS_COMMENTS                   	Top 10 DDA acquisition: Oribitrap Full-Scan at 30k resolving power from 150-2000
MS:MS_COMMENTS                   	m/z. Data-dependent collisional-induced dissociation (CID) fragmentation scans
MS:MS_COMMENTS                   	performed with the LTQ ion trap at 30 normalized collision energy with normal
MS:MS_COMMENTS                   	scan rate.
#MS_METABOLITE_DATA
MS_METABOLITE_DATA:UNITS	Presence/Absence
MS_METABOLITE_DATA_START
Samples	50D-Gly/0_1-neg	50D-Gly/0_2-neg	50D-Gly/0_3-neg	50D-Gly/100_1-neg	50D-Gly/100_2-neg	50D-Gly/100_3-neg	D-Glu/0_1-neg	D-Glu/0_2-neg	D-Glu/0_3-neg	D-Glu/100-neg_tech-rep1	D-Glu/100-neg_tech-rep2	D-Gly/0_1-neg	D-Gly/0_2-neg	D-Gly/0_3-neg	D-Gly/100_1-neg	D-Gly/100_2-neg	D-Gly/100_3-neg	H-Glu/100-neg_tech-rep1	H-Glu/100-neg_tech-rep2	H-Glu/25_1-neg	H-Glu/25_2-neg	H-Glu/25_3-neg	H-Glu/50_1-neg	H-Glu/50_2-neg	H-Glu/50_3-neg	H-Glu/75_1-neg	H-Glu/75_2-neg	H-Glu/75_3-neg	H-Glu/75_repeat_1-neg	H-Glu/75_repeat_2-neg	H-Glu/75_repeat_3-neg	H-Gly/0-neg_tech-rep1	H-Gly/0-neg_tech-rep2	H-Gly/100_1-neg	H-Gly/100_2-neg	H-Gly/100_3-neg	H-Gly/25-neg_tech-rep1	H-Gly/25-neg_tech-rep2	H-Gly/50-neg_tech-rep1	H-Gly/50-neg_tech-rep2	H-Gly/75-neg_tech-rep1	H-Gly/75-neg_tech-rep2
Factors	Carbon source:H-Glucose | Percent D2O:100 | MS ionization mode:negative	Carbon source:H-Glucose | Percent D2O:100 | MS ionization mode:negative	Carbon source:H-Glucose | Percent D2O:100 | MS ionization mode:negative	Carbon source:D-Glucose | Percent D2O:100 | MS ionization mode:negative	Carbon source:D-Glucose | Percent D2O:100 | MS ionization mode:negative	Carbon source:D-Glucose | Percent D2O:100 | MS ionization mode:negative	Carbon source:H-Glucose | Percent D2O:25 | MS ionization mode:negative	Carbon source:H-Glucose | Percent D2O:25 | MS ionization mode:negative	Carbon source:H-Glucose | Percent D2O:25 | MS ionization mode:negative	Carbon source:D-Glucose | Percent D2O:0 | MS ionization mode:negative	Carbon source:D-Glucose | Percent D2O:0 | MS ionization mode:negative	Carbon source:H-Glucose | Percent D2O:50 | MS ionization mode:negative	Carbon source:H-Glucose | Percent D2O:50 | MS ionization mode:negative	Carbon source:H-Glucose | Percent D2O:50 | MS ionization mode:negative	Carbon source:H-Glucose | Percent D2O:75 | MS ionization mode:negative	Carbon source:H-Glucose | Percent D2O:75 | MS ionization mode:negative	Carbon source:H-Glucose | Percent D2O:75 | MS ionization mode:negative	Carbon source:D-Glycerol | Percent D2O:100 | MS ionization mode:negative	Carbon source:D-Glycerol | Percent D2O:100 | MS ionization mode:negative	Carbon source:50/50 H/D-Glycerol | Percent D2O:0 | MS ionization mode:negative	Carbon source:50/50 H/D-Glycerol | Percent D2O:0 | MS ionization mode:negative	Carbon source:50/50 H/D-Glycerol | Percent D2O:0 | MS ionization mode:negative	Carbon source:D-Glycerol | Percent D2O:0 | MS ionization mode:negative	Carbon source:D-Glycerol | Percent D2O:0 | MS ionization mode:negative	Carbon source:D-Glycerol | Percent D2O:0 | MS ionization mode:negative	Carbon source:50/50 H/D-Glycerol | Percent D2O:100 | MS ionization mode:negative	Carbon source:50/50 H/D-Glycerol | Percent D2O:100 | MS ionization mode:negative	Carbon source:50/50 H/D-Glycerol | Percent D2O:100 | MS ionization mode:negative	Carbon source:50/50 H/D-Glycerol | Percent D2O:100 | MS ionization mode:negative	Carbon source:50/50 H/D-Glycerol | Percent D2O:100 | MS ionization mode:negative	Carbon source:50/50 H/D-Glycerol | Percent D2O:100 | MS ionization mode:negative	Carbon source:H-Glycerol | Percent D2O:0 | MS ionization mode:negative	Carbon source:H-Glycerol | Percent D2O:0 | MS ionization mode:negative	Carbon source:H-Glycerol | Percent D2O:100 | MS ionization mode:negative	Carbon source:H-Glycerol | Percent D2O:100 | MS ionization mode:negative	Carbon source:H-Glycerol | Percent D2O:100 | MS ionization mode:negative	Carbon source:H-Glycerol | Percent D2O:25 | MS ionization mode:negative	Carbon source:H-Glycerol | Percent D2O:25 | MS ionization mode:negative	Carbon source:H-Glycerol | Percent D2O:50 | MS ionization mode:negative	Carbon source:H-Glycerol | Percent D2O:50 | MS ionization mode:negative	Carbon source:H-Glycerol | Percent D2O:75 | MS ionization mode:negative	Carbon source:H-Glycerol | Percent D2O:75 | MS ionization mode:negative
PG 30:1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1
PG 30:0	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1
PG 31:1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1
PG 32:1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1
PG 32:0	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1
PG 33:1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1
PG 34:2	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1
PG 34:1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1
PG 35:2	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1
PG 35:1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1
PG 36:2	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1
PG 37:2	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1
MS_METABOLITE_DATA_END
#METABOLITES
METABOLITES_START
metabolite_name	Adduct	Formula
PG 30:1	[M-H]-	C36H69O10P
PG 30:0	[M-H]-	C36H71O10P
PG 31:1	[M-H]-	C37H71O10P
PG 32:1	[M-H]-	C38H73O10P
PG 32:0	[M-H]-	C38H75O10P
PG 33:1	[M-H]-	C39H75O10P
PG 34:2	[M-H]-	C40H75O10P
PG 34:1	[M-H]-	C40H77O10P
PG 35:2	[M-H]-	C41H77O10P
PG 35:1	[M-H]-	C41H79O10P
PG 36:2	[M-H]-	C42H79O10P
PG 37:2	[M-H]-	C43H81O10P
METABOLITES_END
#END