#METABOLOMICS WORKBENCH araskind_20170802_132817 DATATRACK_ID:1182 STUDY_ID:ST000833 ANALYSIS_ID:AN001843 PROJECT_ID:PR000594 VERSION 1 CREATED_ON January 16, 2019, 9:11 am #PROJECT PR:PROJECT_TITLE Influence of murine norovirus on cell metabolism PR:PROJECT_TYPE MS analysis PR:PROJECT_SUMMARY C6 effects in RAW PR:INSTITUTE University of Michigan PR:DEPARTMENT Microbiology & Immunology PR:LABORATORY Passalacqua Lab PR:LAST_NAME Passalacqua PR:FIRST_NAME Karla PR:ADDRESS Ann Arbor, MI PR:EMAIL kpassal@umich.edu PR:PHONE 734-763-5639 #STUDY ST:STUDY_TITLE Influence of murine norovirus on cell metabolism ST:STUDY_TYPE MS analysis ST:STUDY_SUMMARY 6.5 X 10^6 cells were plated in 10 cm Tissue Culture dishes and allowed to ST:STUDY_SUMMARY recover overnight at 37 degrees and 5% CO2. In the am, supernatant was removed, ST:STUDY_SUMMARY and 12 mL of medium containing either Murine Norovirus-1 (MNV) at an MOI=5 or ST:STUDY_SUMMARY medium containing a v/v match of mock lysate was added to the cells. Plates were ST:STUDY_SUMMARY rocked on ice for 1 hour, then cells were washed 3X with cold DPBS++, and plain ST:STUDY_SUMMARY medium as added to cells. Plates were then incubated for 7.5 hours at 37 degrees ST:STUDY_SUMMARY and 5% CO2. Cells were washed with 12 mL of 150 mM Ammonium Acetate, swirled 8 ST:STUDY_SUMMARY times, and immediately quenched with liquid nitrogen. Cells were then frozen at ST:STUDY_SUMMARY -80 degrees. ST:INSTITUTE University of Michigan ST:DEPARTMENT Biomedical Research Core Facilities ST:LABORATORY Metabolomics core ST:LAST_NAME Kachman ST:FIRST_NAME Maureen ST:ADDRESS 6300 Brehm Tower, 1000 Wall Street, Ann Arbor, MI 48105-5714 ST:EMAIL mkachman@med.umich.edu ST:PHONE (734) 232-8175 ST:NUM_GROUPS 2 ST:TOTAL_SUBJECTS 1 ST:STUDY_COMMENTS BIOHAZARD - MURINE NOROVIRUS - SHOULD BE ENTIRELY NON-INFECTIOUS DUE TO THE ST:STUDY_COMMENTS LIQUID NITROGEN, BUT PLEASE HANDLE AND DISPOSE OF IN BIOHAZARD WASTE. IMPORTANT ST:STUDY_COMMENTS NOTE: I request that the metabolites of the TCA+ assay that are to be assessed ST:STUDY_COMMENTS quantitatively please be sure to include ATP, ADP, AMP, NAD, NADH, NADP, NADPH, ST:STUDY_COMMENTS E4P, S7P, 6PG, G3P H6P and Lactate. Also, I would request that both reduced and ST:STUDY_COMMENTS oxidized Glutathione as well as Fructose 1,6-bisphosphate be included in the ST:STUDY_COMMENTS non-quantitative data. #SUBJECT SU:SUBJECT_TYPE MOUSE SU:SUBJECT_SPECIES Mus musculus SU:TAXONOMY_ID 10090 #SUBJECT_SAMPLE_FACTORS: SUBJECT(optional)[tab]SAMPLE[tab]FACTORS(NAME:VALUE pairs separated by |)[tab]Additional sample data SUBJECT_SAMPLE_FACTORS SU0015382 S00023719 Medium:mock lysate SUBJECT_SAMPLE_FACTORS SU0015382 S00023720 Medium:mock lysate SUBJECT_SAMPLE_FACTORS SU0015382 S00023721 Medium:mock lysate SUBJECT_SAMPLE_FACTORS SU0015382 S00023722 Medium:MNV infected SUBJECT_SAMPLE_FACTORS SU0015382 S00023723 Medium:MNV infected SUBJECT_SAMPLE_FACTORS SU0015382 S00023724 Medium:MNV infected #COLLECTION CO:COLLECTION_SUMMARY - CO:SAMPLE_TYPE Cultured cells #TREATMENT TR:TREATMENT_SUMMARY - #SAMPLEPREP SP:SAMPLEPREP_SUMMARY - SP:SAMPLEPREP_PROTOCOL_FILENAME A011_Glycolysis-TCA-nucleotides.pdf #CHROMATOGRAPHY CH:CHROMATOGRAPHY_TYPE HILIC CH:INSTRUMENT_NAME Agilent CH:COLUMN_NAME Phenomenex Luna NH2 (150 x 1mm, 3um) #ANALYSIS AN:ANALYSIS_TYPE MS AN:ANALYSIS_PROTOCOL_FILE A043-TCA-Plus-20170713.pdf AN:ACQUISITION_PARAMETERS_FILE QTOF-002-HILIC-35min-1mm.m #MS MS:INSTRUMENT_NAME Agilent 6520B QTOF MS:INSTRUMENT_TYPE QTOF MS:MS_TYPE ESI MS:ION_MODE POSITIVE MS:MS_COMMENTS none #MS_METABOLITE_DATA MS_METABOLITE_DATA:UNITS counts MS_METABOLITE_DATA_START Samples S00023719 S00023720 S00023721 S00023722 S00023723 S00023724 Factors Medium:mock lysate Medium:mock lysate Medium:mock lysate Medium:MNV infected Medium:MNV infected Medium:MNV infected GLUCONIC ACID 231272.8325 195366.8393 135925.3264 418134.4743 222479.7657 182665.4254 TAURINE 63950932.64 50676058.84 56377501.2 64958587.51 60022849.55 65321672.22 URIC ACID 128505.5775 176650.1692 251363.785 188160.977 199117.0079 213234.4661 XANTHINE 397154.9991 2930716.691 3668065.387 826107.3627 1530321.375 1783656.152 D-ERYTHROSE 4-PHOSPHATE 68253.34949 230475.7349 172808.9529 70034.44742 96794.80692 81274.44164 GLUTATHIONE 1333581.464 2056812.267 1561429 1064190.265 1584014.249 1362084.82 2'-DEOXYURIDINE 7253344.656 8946914.424 6844326.59 9380767.774 9798070.569 7643628.16 L-PROLINE 3767138.527 3132992.284 2325280.908 3887644.334 3299241.669 2485465.443 UDP-GLUCURONIC ACID 232592.6013 190078.3693 184857.6334 271535.8187 208423.3675 207507.5171 ACETYLPHOSPHATE 91244.66741 78916.12187 69523.97281 70168.05561 60598.3589 63776.0928 ADENINE 1164772.548 1246252.849 540740.9672 1393793.209 942504.5871 1139395.996 L-GLUTAMIC ACID 24347135 21428597.35 18020902.33 27126850.77 23862628.67 17076423.03 N-ACETYL-D-GLUCOSAMINE 1-PHOSPHATE 312522.7031 369610.8452 422483.7269 416423.9878 385810.3884 362591.6729 OLEIC ACID 5410348.135 11522826.79 8355434.393 3952359.649 6940619.419 7706069.348 UDP-N-ACETYLGLUCOSAMINE 4292583.078 3179002.061 2598179.263 4054677.535 3737952.988 3057997.671 FAD 72841.18803 42676.72681 27118.7051 57933.30811 41211.39655 47903.06924 STEARIC ACID 2965954.06 4651451.499 6114286.066 2036730.769 3623317.783 3218775.518 UDPGLUCOSE 6813266.267 4435100.312 3654133.488 7245489.001 5427123.895 5047160.341 D-GLUCOSE 16253130.07 19251637.41 17248330.5 19513338.62 19736630.62 17592831.86 CREATINE 2395975.714 2141465.872 1918906.88 2906106.758 2631940.971 1919562.431 CREATININE 297219.2419 374528.4053 311798.0143 506759.7217 373652.8791 320213.4064 L-ALANINE 3774448.324 3454991.492 3114569.297 4732466.514 4063465.068 3260791.313 ROBISON ESTER 1626588.401 503137.2617 851130.7059 1573490.932 1015591.661 1220485.462 L-ASPARTIC ACID 7663909.607 6275688.698 6397646.055 8866172.921 8024132.699 6295980.616 L-GLUTAMINE 55058705.62 52512792.09 42095211.96 61563477.72 58594616.31 46958160.98 L-LYSINE 1801621.082 2126100.128 2083204.955 1957343.727 2248878.242 1829795.268 INOSINE 164571.7201 1188712.099 1963074.757 431494.5699 589238.7882 520082.4848 URIDYLIC ACID 43170.14485 4165086.891 3825391.127 940703.6144 1067595.452 1132324.982 URIDINE 5'-DIPHOSPHATE 6403716.934 9231491.483 11083000.62 14104370.52 12092385.04 12926972.75 5'-CYTIDYLIC ACID 190875.1604 830869.53 839053.2156 297778.204 329598.6036 368011.7457 URIDINE 5'-TRIPHOSPHATE 2621230.184 967566.3307 10539996.95 5065761.362 4533698.236 7581358.954 L-METHIONINE 1547890.779 1742222.979 1665727.071 1655607.327 1895202.334 1324408.34 L-PHENYLALANINE 13824086.46 15048589.86 13210024.42 14734628.93 14641754.82 12952189.94 DL-SERINE 2012613.679 2051413.385 1819297.063 2519794.112 1969684.961 1988578.614 L-HISTIDINE 651691.6843 639077.7877 541138.0872 435896.7859 888090.5051 436770.5541 L-VALINE 55920306.18 47779215.79 50967567.02 58642020.92 52665181.05 58163471.29 L-THREONINE 4670444.952 4657878.961 3795190.004 5193380.832 4723426.337 3757100.062 L-TRYPTOPHAN 2808472.591 2860491.255 2510975.15 2714631.986 2696030.843 2518692.659 L-ARGININE 443056.5116 376419.2049 362579.5899 515893.1147 411769.8499 377442.5514 FLAVIN MONONUCLEOTIDE 24924.60098 31038.19395 20998.92372 17530.79977 20447.0685 18891.2144 OXIGLUTATIONE 44046113.39 33512019.1 22734260.76 36888934.8 33663105.86 25179734.61 DIHYDROXYACETONE PHOSPHATE 1537595.409 1421916.477 608768.7415 3408022.609 1041532.855 680608.7347 5'-GUANYLIC ACID 29213.1167 565806.3225 1466594.598 56482.55216 166099.5232 205492.716 O-PHOSPHO-L-SERINE 4229918.202 3968304.77 2911031.633 4268050.316 3738432.049 3887825.529 INOSINIC ACID 282333.2222 1206546.165 1069432.503 342528.8279 410595.285 384180.0123 GUANOSINE DIPHOSPHATE 1271817.678 2156483.596 1714626.48 2130085.923 2272047.605 2267554.747 PHOSPHOCREATINE 392626.5374 5491.504873 4478.328072 223191.3739 20505.27198 30866.82968 L-CITRULLINE 59952.41496 100631.5741 73477.27043 60058.67222 102943.2835 51894.27635 PALMITIC ACID 5319292.937 7321385.209 6875942.049 3689921.481 5613408.173 5851126.878 MS_METABOLITE_DATA_END #METABOLITES METABOLITES_START metabolite_name moverz_quant ri ri_type pubchem_id inchi_key kegg_id other_id other_id_type GLUCONIC ACID 10690 TAURINE 1123 URIC ACID 1175 XANTHINE 1188 D-ERYTHROSE 4-PHOSPHATE 122357 GLUTATHIONE 124886 2'-DEOXYURIDINE 13712 L-PROLINE 145742 UDP-GLUCURONIC ACID 17473 ACETYLPHOSPHATE 186 ADENINE 190 L-GLUTAMIC ACID 33032 N-ACETYL-D-GLUCOSAMINE 1-PHOSPHATE 440272 OLEIC ACID 445639 UDP-N-ACETYLGLUCOSAMINE 445675 FAD 46906035 STEARIC ACID 5281 UDPGLUCOSE 53477679 D-GLUCOSE 5793 CREATINE 586 CREATININE 588 L-ALANINE 5950 ROBISON ESTER 5958 L-ASPARTIC ACID 5960 L-GLUTAMINE 5961 L-LYSINE 5962 INOSINE 6021 URIDYLIC ACID 6030 URIDINE 5'-DIPHOSPHATE 6031 5'-CYTIDYLIC ACID 6131 URIDINE 5'-TRIPHOSPHATE 6133 L-METHIONINE 6137 L-PHENYLALANINE 6140 DL-SERINE 617 L-HISTIDINE 6274 L-VALINE 6287 L-THREONINE 6288 L-TRYPTOPHAN 6305 L-ARGININE 6322 FLAVIN MONONUCLEOTIDE 643976 OXIGLUTATIONE 65359 DIHYDROXYACETONE PHOSPHATE 668 5'-GUANYLIC ACID 6804 O-PHOSPHO-L-SERINE 68841 INOSINIC ACID 8582 GUANOSINE DIPHOSPHATE 8977 PHOSPHOCREATINE 9548602 L-CITRULLINE 9750 PALMITIC ACID 985 METABOLITES_END #END