#METABOLOMICS WORKBENCH mososa4177_20190425_053345 DATATRACK_ID:1711 STUDY_ID:ST001177 ANALYSIS_ID:AN001953 PROJECT_ID:PR000789
VERSION             	1
CREATED_ON             	May 1, 2019, 2:15 pm
#PROJECT
PR:PROJECT_TITLE                 	Short Chain Lipid Project
PR:PROJECT_TYPE                  	Untargted analysis
PR:PROJECT_SUMMARY               	Detect phosphatidylcholine (PC) with short acyl chains on the Golgi membrane
PR:INSTITUTE                     	Brigham and Women’s Hospital
PR:DEPARTMENT                    	Division of Rheumatology, Immunology and Allergy
PR:LAST_NAME                     	Hsu
PR:FIRST_NAME                    	Victor
PR:ADDRESS                       	60 Fenwood Road, Boston, MA 02115 USA
PR:EMAIL                         	vhsu@bwh.harvard.edu
PR:PHONE                         	617-525-1103
#STUDY
ST:STUDY_TITLE                   	Analysis of short chain phosphatidylcholine (PC) on the Golgi membrane
ST:STUDY_TYPE                    	Golgi membrane lipids characterization
ST:STUDY_SUMMARY                 	Studies on vesicle formation by the Coat Protein I (COPI) complex have
ST:STUDY_SUMMARY                 	contributed to a basic understanding of how vesicular transport is initiated. We
ST:STUDY_SUMMARY                 	have identified that short chain lipids promote membrane properties that are
ST:STUDY_SUMMARY                 	conducive for fission. Here we investigated short chain PCs on Golgi membrane.
ST:STUDY_SUMMARY                 	These findings will advance the understanding of how lipid geometry contributes
ST:STUDY_SUMMARY                 	to membrane deformation needed for vesicle fission.
ST:INSTITUTE                     	University of Kentucky
ST:LAST_NAME                     	Morris
ST:FIRST_NAME                    	Andrew
ST:ADDRESS                       	900 S limestone
ST:EMAIL                         	a.j.morris@uky.edu
ST:PHONE                         	859-323-3749
#SUBJECT
SU:SUBJECT_TYPE                  	Other
SU:SUBJECT_SPECIES               	Cricetulus griseus
SU:TAXONOMY_ID                   	10029
#FACTORS
#SUBJECT_SAMPLE_FACTORS:         	SUBJECT(optional)[tab]SAMPLE[tab]FACTORS(NAME:VALUE pairs separated by |)[tab]Additional sample data
SUBJECT_SAMPLE_FACTORS           	-	20180922_sample_01	Sample Type:Golgi membrane sample	
#COLLECTION
CO:COLLECTION_SUMMARY            	Golgi membrane were prepared from Chinese hamster ovary cell line as described
CO:COLLECTION_SUMMARY            	previously. J Cell Biol 159, 69-78 (2002); EMBO J 24, 4133-4143 (2005).
CO:SAMPLE_TYPE                   	CHO cells
#TREATMENT
TR:TREATMENT_SUMMARY             	Golgi membrane were prepared as described previously. J Cell Biol 159, 69-78
TR:TREATMENT_SUMMARY             	(2002); EMBO J 24, 4133-4143 (2005).
#SAMPLEPREP
SP:SAMPLEPREP_SUMMARY            	5 mg of Golgi membrane fraction was extracted using acidified CHCl3/methanol
SP:SAMPLEPREP_SUMMARY            	(1:2)
#CHROMATOGRAPHY
CH:CHROMATOGRAPHY_SUMMARY        	Lipid extracts were separated on a Waters ACQUITY BEH C8 column (2.1 × 100 mm,
CH:CHROMATOGRAPHY_SUMMARY        	1.7 μm) with the temperature maintained at 40 °C. The flow rate was 250
CH:CHROMATOGRAPHY_SUMMARY        	μL/min, and the mobile phases were consisted of 60:40 water/acetonitrile (A),
CH:CHROMATOGRAPHY_SUMMARY        	and 90:10 isopropanol/acetonitrile (B), both containing 10 mM ammonium formate
CH:CHROMATOGRAPHY_SUMMARY        	and 0.1% formic acid. The samples were eluted with a linear gradient from 5 % B
CH:CHROMATOGRAPHY_SUMMARY        	to 97 % B over 20 min, maintained at 97 % B for 4 min and re-equilibration with
CH:CHROMATOGRAPHY_SUMMARY        	5 % B for 6 min. The sample injection volume was 5 μL
CH:CHROMATOGRAPHY_TYPE           	Reversed phase
MS:INSTRUMENT_NAME               	Thermo Q Exactive Orbitrap
CH:COLUMN_NAME                   	ACQUITY BEH C8 column (2.1 × 100 mm, 1.7 µm)
#ANALYSIS
AN:ANALYSIS_TYPE                 	MS
#MS
MS:INSTRUMENT_NAME               	Thermo Q Exactive Orbitrap
MS:INSTRUMENT_TYPE               	Orbitrap
MS:MS_TYPE                       	ESI
MS:ION_MODE                      	POSITIVE
MS:MS_COMMENTS                   	Data analysis and accurate mass calculation were performed using the software
MS:MS_COMMENTS                   	Xcalibur 4.0
MS:MS_RESULTS_FILE               	ST001177_AN001953_Results.txt	UNITS:Peak area	Has m/z:Yes	Has RT:Yes	RT units:Minutes
#END