#METABOLOMICS WORKBENCH araskind_20170802_184250 DATATRACK_ID:1186 STUDY_ID:ST000835 ANALYSIS_ID:AN001336 PROJECT_ID:PR000595 VERSION 1 CREATED_ON August 9, 2017, 9:44 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 Deubiquitinase inhibitor compound C6 effect on cellular metabolism ST:STUDY_TYPE MS analysis ST:STUDY_SUMMARY 6.0 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 fresh medium with either 2.5 micromolar Compound C6 or a v/v match ST:STUDY_SUMMARY of DMSO (vehicle control) was added to the cells. Cells were incubated at 37 ST:STUDY_SUMMARY degrees in 5% CO2 for 30 minutes. Cells were washed 3X with cold DPBS++ and 10 ST:STUDY_SUMMARY mL of medium was added to the cells with either 1.0 micromolar Compound C6 or ST:STUDY_SUMMARY v/v match of DMSO. Cells were incubated at 37 degrees and 5% CO2 for 7.0 hours. ST:STUDY_SUMMARY Cells were washed with 10 mL of 150 mM Ammonium Acetate, swirled, washed again ST:STUDY_SUMMARY with 5 mL, and immediately quenched with liquid nitrogen. Cells were then frozen ST:STUDY_SUMMARY at -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 7 ST:TOTAL_SUBJECTS 1 ST:STUDY_COMMENTS 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 in particular. Also, I would request that ST:STUDY_COMMENTS both reduced and oxidized Glutathione as well as Fructose 1,6-bisphosphate be ST:STUDY_COMMENTS included in the 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 SU0016447 S00026310 Sample type:Cell Line | Treatment:Vehicle | Type:RAW SUBJECT_SAMPLE_FACTORS SU0016447 S00026311 Sample type:Cell Line | Treatment:Vehicle | Type:RAW SUBJECT_SAMPLE_FACTORS SU0016447 S00026312 Sample type:Cell Line | Treatment:Vehicle | Type:RAW SUBJECT_SAMPLE_FACTORS SU0016447 S00026313 Sample type:Cell Line | Treatment:Vehicle | Type:RAW SUBJECT_SAMPLE_FACTORS SU0016447 S00026314 Sample type:Cell Line | Treatment:Vehicle | Type:RAW SUBJECT_SAMPLE_FACTORS SU0016447 S00026315 Sample type:Cell Line | Treatment:DUB inhibitor | Type:RAW SUBJECT_SAMPLE_FACTORS SU0016447 S00026316 Sample type:Cell Line | Treatment:DUB inhibitor | Type:RAW SUBJECT_SAMPLE_FACTORS SU0016447 S00026317 Sample type:Cell Line | Treatment:DUB inhibitor | Type:RAW SUBJECT_SAMPLE_FACTORS SU0016447 S00026318 Sample type:Cell Line | Treatment:DUB inhibitor | Type:RAW SUBJECT_SAMPLE_FACTORS SU0016447 S00026319 Sample type:Cell Line | Treatment:DUB inhibitor | Type:RAW SUBJECT_SAMPLE_FACTORS SU0016447 S00026320 Sample type:Macrophages | Treatment:Control-Other_Cells | Type:Primary BL6 BMM SUBJECT_SAMPLE_FACTORS SU0016447 S00026321 Sample type:Macrophages | Treatment:Control-Other_Cells | Type:Primary BL6 BMM #COLLECTION CO:COLLECTION_SUMMARY - #TREATMENT TR:TREATMENT_SUMMARY - #SAMPLEPREP SP:SAMPLEPREP_SUMMARY - SP:SAMPLEPREP_PROTOCOL_FILENAME A043-TCA-Plus-20170713.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 #MS MS:MS_COMMENTS - MS:INSTRUMENT_NAME Agilent 6520B QTOF MS:INSTRUMENT_TYPE QTOF MS:MS_TYPE ESI MS:ION_MODE NEGATIVE #MS_METABOLITE_DATA MS_METABOLITE_DATA:UNITS ISTD response ratio MS_METABOLITE_DATA_START Samples S00026310 S00026311 S00026312 S00026313 S00026314 S00026315 S00026316 S00026317 S00026318 S00026319 S00026320 S00026321 Factors Sample type:Cell Line | Treatment:Vehicle | Type:RAW Sample type:Cell Line | Treatment:Vehicle | Type:RAW Sample type:Cell Line | Treatment:Vehicle | Type:RAW Sample type:Cell Line | Treatment:Vehicle | Type:RAW Sample type:Cell Line | Treatment:Vehicle | Type:RAW Sample type:Cell Line | Treatment:DUB inhibitor | Type:RAW Sample type:Cell Line | Treatment:DUB inhibitor | Type:RAW Sample type:Cell Line | Treatment:DUB inhibitor | Type:RAW Sample type:Cell Line | Treatment:DUB inhibitor | Type:RAW Sample type:Cell Line | Treatment:DUB inhibitor | Type:RAW Sample type:Macrophages | Treatment:Control-Other_Cells | Type:Primary BL6 BMM Sample type:Macrophages | Treatment:Control-Other_Cells | Type:Primary BL6 BMM TAURINE 49.56811707 48.54949698 47.90172398 57.42753038 51.11318861 37.65909321 48.07485695 37.76207842 52.97245757 51.6370897 32.85490365 31.94544117 GLUTATHIONE 24.41032857 23.91276192 20.11038678 36.29528681 34.1884367 29.5369422 41.61692386 33.3851043 40.45613771 51.13668789 5.116975783 3.579148851 L-PROLINE .648977508 .588147746 .525668834 .769359161 .731922677 .39737358 .58749379 .348065237 .638699408 .575330099 .09074017 .093175441 UDP-GLUCURONIC ACID .250717658 .235679473 .218093573 .316164807 .251943241 .131233638 .179530554 .123658727 .221955532 .216910784 .083083539 .068454739 L-GLUTAMIC ACID 12.59342375 12.84708597 11.14399219 16.29029545 13.91941551 6.211049971 7.61351969 5.02873064 8.843073933 8.781932453 2.186146978 2.089266616 OLEIC ACID 4.003163662 6.065301128 3.503810997 5.309568895 4.207848767 4.498074956 4.004760451 3.308933898 3.733666098 4.084069369 1.820054692 1.901186869 UDP-N-ACETYLGLUCOSAMINE 2.74207697 3.044655744 1.958948081 3.197225701 3.186864106 1.513814249 2.644571328 1.478100987 2.834655445 2.732038754 .67935663 .685343394 STEARIC ACID 13.90541875 7.559989292 10.72056432 19.6714425 12.56597582 8.824153647 9.751716993 8.615595773 12.93525669 10.115417 10.26984689 10.61044932 UDPGLUCOSE 2.613033394 2.464030186 2.457564229 3.318826942 3.07410661 1.523208578 2.453854103 1.751242896 3.018897601 2.783798019 .574203062 .559020954 L-ASPARTIC ACID 2.567211529 2.611561844 2.329361898 3.494352988 2.977435047 1.533078897 1.916731333 1.100245584 2.08 2.092204052 .128372063 .117845726 L-GLUTAMINE 21.793366 21.10406853 18.0088992 28.06698364 25.59309846 13.32309089 17.70765022 11.70799645 19.33497234 18.11394402 2.948344805 3.171742037 INOSINE .776228832 .932822329 .711855481 .872627133 .785547261 .404439125 .668662884 .551844916 .576838693 1.083417112 6.742144122 6.033233257 URIDYLIC ACID .312985262 .492723741 .351103406 .394248159 .343774011 .136383341 .17822471 .209548642 .209217039 .240601315 .388182444 .381628214 URIDINE 5'-DIPHOSPHATE 1.888592337 1.905884541 1.842549061 2.210726121 1.896706633 .796181895 1.195290075 .990276246 1.507654136 1.353470039 .346771314 .329816961 L-TYROSINE 1.150641826 1.185520006 .97823821 1.276913473 1.196389131 .74077314 1.258278046 .731465005 1.283164473 1.107432632 .402748576 .491118105 5'-CYTIDYLIC ACID .100917296 .16225549 .097593297 .111074216 .104665649 .06798727 .05527772 .081446361 .115886552 .110460445 .097047611 .093386884 L-PHENYLALANINE 1.173299587 1.28403304 1.012872086 1.478845133 1.285288439 .937611125 1.274133509 .80497155 1.333888628 1.105547031 .393708103 .482496857 L-VALINE 11.86104536 12.13944769 12.10906386 13.23140816 11.85919515 12.49822507 12.76583665 12.09563283 12.23601942 12.04652818 11.43385614 11.39776137 L-ISOLEUCINE 4.535578158 4.791291508 3.883167749 5.198377309 4.999907135 3.57170116 4.293244824 2.8833472 4.521104541 3.867759319 1.389598861 1.583492662 L-ARGININE .388121044 .401580534 .303201304 .48501583 .378286566 .458219716 .418949324 .234569586 .414558714 .344874323 .008606638 .004029971 5'-GUANYLIC ACID .167469049 .26369193 .184437459 .181749994 .167222404 .092038705 .131087374 .116580011 .155818309 .157877594 .402387495 .396364133 O-PHOSPHO-L-SERINE 1.115217549 1.150115921 .950627518 1.440922409 1.297995427 .556662267 .588875901 .289372071 .602423783 .78298197 .060093879 .048053512 INOSINIC ACID .02614785 .04559122 .011491668 .02210074 .009544663 .005460923 .006089353 .084122699 .01437604 .009783372 .370509918 .3644436 GUANOSINE DIPHOSPHATE .903856516 1.068165169 .808073655 1.204785312 1.142199893 .585364731 .996323216 .697966489 .909238003 .965528463 .51655979 .485970693 PHOSPHOCREATINE .024908294 .017810527 .017369678 .014744525 .015477335 .007249382 .026136643 .015538026 .02199351 .021161417 .041148994 .04431882 L-GLUTATHIONE (OXIDIZED) 8.444317079 7.203448848 5.975101954 7.924607464 6.534507383 4.227275069 10.81866439 6.217915258 14.86286016 9.768116847 3.800142024 4.257506746 PALMITIC ACID 13.44970078 8.359622357 10.6753035 18.78137757 13.2908155 8.341207615 10.74124859 9.592569478 13.75945308 10.05600124 9.471307633 9.426362114 PC(34:2) .104935439 .173810236 .197569878 .255148024 .167874637 .297700782 .213066428 .175690898 .032975026 .201958444 .333684738 .162426631 PI(38:4) .256480186 .764059834 .836517295 1.066864008 .758323646 2.303288787 1.243440141 .860660447 .045659056 1.211581556 1.942890931 .76979881 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 TAURINE 1123 GLUTATHIONE 124886 L-PROLINE 145742 UDP-GLUCURONIC ACID 17473 L-GLUTAMIC ACID 33032 OLEIC ACID 445639 UDP-N-ACETYLGLUCOSAMINE 445675 STEARIC ACID 5281 UDPGLUCOSE 53477679 L-ASPARTIC ACID 5960 L-GLUTAMINE 5961 INOSINE 6021 URIDYLIC ACID 6030 URIDINE 5'-DIPHOSPHATE 6031 L-TYROSINE 6057 5'-CYTIDYLIC ACID 6131 L-PHENYLALANINE 6140 L-VALINE 6287 L-ISOLEUCINE 6306 L-ARGININE 6322 5'-GUANYLIC ACID 6804 O-PHOSPHO-L-SERINE 68841 INOSINIC ACID 8582 GUANOSINE DIPHOSPHATE 8977 PHOSPHOCREATINE 9548602 L-GLUTATHIONE (OXIDIZED) 975 PALMITIC ACID 985 PC(34:2) PC(34:2) REFMET PI(38:4) PI(38:4) REFMET METABOLITES_END #END