#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