#METABOLOMICS WORKBENCH araskind_20150409_9167301_mwtab.txt DATATRACK_ID:227 STUDY_ID:ST000153 ANALYSIS_ID:AN000244 PROJECT_ID:PR000129 VERSION 1 CREATED_ON 2016-09-17 #PROJECT PR:PROJECT_TITLE Metabolic effects of Noxa PR:PROJECT_SUMMARY labeled glutamine metabolomics on Noxa expressing cells PR:INSTITUTE University of Minnesota PR:DEPARTMENT Masonic Cancer Center PR:LABORATORY Kelekar Lab PR:LAST_NAME Kelekar PR:FIRST_NAME Ameeta PR:ADDRESS 420 Delaware Street SE, Minneapolis, MN 55455 PR:EMAIL ameeta@umn.edu PR:PHONE 612-626-2358 #STUDY ST:STUDY_TITLE Primary T Cell Noxa Knockdown (Donor 8)-II ST:STUDY_TYPE See attached document ST:STUDY_SUMMARY Labeling of cells was carried out in triplicate with each sample containing ST:STUDY_SUMMARY Cells were deprived of glucose or glutamine for 1 h then resuspended in 10mM ST:STUDY_SUMMARY D-glucose labeled or 4 mM [13C5]-U-glutamine labeled medium, respectively. At ST:STUDY_SUMMARY time cells were also activated with 5 ug/ml anti-CD3 and anti-CD28. Cells were ST:STUDY_SUMMARY (and activated) for 16 h when samples were spun down, and cells were ST:STUDY_SUMMARY in 200 ul ice cold methanol. All samples were immediately stored in -80°C. 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@umich.edu ST:PHONE - ST:NUM_GROUPS 3 ST:TOTAL_SUBJECTS 16 #SUBJECT SU:SUBJECT_TYPE - SU:SUBJECT_SPECIES Homo sapiens SU:TAXONOMY_ID 9606 SU:SUBJECT_COMMENTS 16 subjects #SUBJECT_SAMPLE_FACTORS: SUBJECT(optional)[tab]SAMPLE[tab]FACTORS(NAME:VALUE pairs separated by |)[tab]Additional sample data SUBJECT_SAMPLE_FACTORS SU0011714 S00017304 Glucose label:no | Glutamine label:no SUBJECT_SAMPLE_FACTORS SU0011714 S00017312 Glucose label:no | Glutamine label:no SUBJECT_SAMPLE_FACTORS SU0011708 S00017298 Glucose label:no | Glutamine label:yes SUBJECT_SAMPLE_FACTORS SU0011709 S00017299 Glucose label:no | Glutamine label:yes SUBJECT_SAMPLE_FACTORS SU0011710 S00017300 Glucose label:no | Glutamine label:yes SUBJECT_SAMPLE_FACTORS SU0011711 S00017301 Glucose label:no | Glutamine label:yes SUBJECT_SAMPLE_FACTORS SU0011712 S00017302 Glucose label:no | Glutamine label:yes SUBJECT_SAMPLE_FACTORS SU0011713 S00017303 Glucose label:no | Glutamine label:yes SUBJECT_SAMPLE_FACTORS SU0011715 S00017305 Glucose label:no | Glutamine label:yes SUBJECT_SAMPLE_FACTORS SU0011722 S00017313 Glucose label:no | Glutamine label:yes SUBJECT_SAMPLE_FACTORS SU0011716 S00017306 Glucose label:yes | Glutamine label:no SUBJECT_SAMPLE_FACTORS SU0011717 S00017307 Glucose label:yes | Glutamine label:no SUBJECT_SAMPLE_FACTORS SU0011718 S00017308 Glucose label:yes | Glutamine label:no SUBJECT_SAMPLE_FACTORS SU0011719 S00017309 Glucose label:yes | Glutamine label:no SUBJECT_SAMPLE_FACTORS SU0011720 S00017310 Glucose label:yes | Glutamine label:no SUBJECT_SAMPLE_FACTORS SU0011721 S00017311 Glucose label:yes | Glutamine label:no #COLLECTION CO:COLLECTION_SUMMARY - CO:SAMPLE_TYPE Cells #TREATMENT TR:TREATMENT_SUMMARY - #SAMPLEPREP SP:SAMPLEPREP_SUMMARY - SP:SAMPLEPREP_PROTOCOL_FILENAME Fluxomics_analysis_protocol-2015-03-11.docx #CHROMATOGRAPHY CH:CHROMATOGRAPHY_TYPE HILIC CH:INSTRUMENT_NAME Agilent 1260 CH:COLUMN_NAME Phenomenex Luna NH2 (150 x 1mm, 3um) CH:METHODS_ID AQM010 CH:METHODS_FILENAME QTOF-002-HILIC-35min-1mm-No_Insert.m #ANALYSIS AN:ANALYSIS_TYPE MS AN:LABORATORY_NAME MRC2 (University of Michigan) AN:ACQUISITION_PARAMETERS_FILE QTOF-002-HILIC-35min-1mm.m AN:PROCESSING_PARAMETERS_FILE EX00404-MassHunterQuant-LCMS-Flux-DataAnalysis-Method.m #MS MS:MS_COMMENTS - MS:INSTRUMENT_NAME Agilent 6520 QTOF MS:INSTRUMENT_TYPE QTOF MS:MS_TYPE ESI MS:ION_MODE NEGATIVE #MS_METABOLITE_DATA MS_METABOLITE_DATA:UNITS Area MS_METABOLITE_DATA_START Samples S00017304 S00017304 S00017312 S00017312 S00017298 S00017298 S00017299 S00017299 S00017300 S00017300 S00017301 S00017301 S00017302 S00017302 S00017303 S00017303 S00017305 S00017305 S00017313 S00017313 S00017306 S00017306 S00017307 S00017307 S00017308 S00017308 S00017309 S00017309 S00017310 S00017310 S00017311 S00017311 Factors Glucose label:no | Glutamine label:no Glucose label:no | Glutamine label:no Glucose label:no | Glutamine label:no Glucose label:no | Glutamine label:no Glucose label:no | Glutamine label:yes Glucose label:no | Glutamine label:yes Glucose label:no | Glutamine label:yes Glucose label:no | Glutamine label:yes Glucose label:no | Glutamine label:yes Glucose label:no | Glutamine label:yes Glucose label:no | Glutamine label:yes Glucose label:no | Glutamine label:yes Glucose label:no | Glutamine label:yes Glucose label:no | Glutamine label:yes Glucose label:no | Glutamine label:yes Glucose label:no | Glutamine label:yes Glucose label:no | Glutamine label:yes Glucose label:no | Glutamine label:yes Glucose label:no | Glutamine label:yes Glucose label:no | Glutamine label:yes Glucose label:yes | Glutamine label:no Glucose label:yes | Glutamine label:no Glucose label:yes | Glutamine label:no Glucose label:yes | Glutamine label:no Glucose label:yes | Glutamine label:no Glucose label:yes | Glutamine label:no Glucose label:yes | Glutamine label:no Glucose label:yes | Glutamine label:no Glucose label:yes | Glutamine label:no Glucose label:yes | Glutamine label:no Glucose label:yes | Glutamine label:no Glucose label:yes | Glutamine label:no Alanine_123 0.0000 0.0000 0.0000 942.8683 0.0000 0.0000 0.0000 0.0000 0.0000 0.0000 0.0000 0.0000 0.0000 0.0000 0.0000 0.0000 Alanine_123_13C_1 0.0000 0.0000 0.0000 0.0000 0.0000 0.0000 0.0000 0.0000 0.0000 0.0000 0.0000 0.0000 0.0000 0.0000 0.0000 0.0000 Alanine_123_13C_2 0.0000 0.0000 0.0000 0.0000 0.0000 0.0000 0.0000 0.0000 0.0000 0.0000 0.0000 0.0000 0.0000 0.0000 0.0000 0.0000 Alanine_123_13C_3 0.0000 0.0000 0.0000 0.0000 0.0000 0.0000 0.0000 0.0000 0.0000 0.0000 0.0000 0.0000 0.0000 0.0000 0.0000 0.0000 alpha-Ketoglutarate_12345 0.0000 0.0000 898.3651 0.0000 0.0000 0.0000 0.0000 0.0000 1433.0704 0.0000 0.0000 460.3290 671.4062 0.0000 582.6751 675.0641 alpha-Ketoglutarate_12345_13C_1 0.0000 0.0000 0.0000 0.0000 0.0000 0.0000 0.0000 0.0000 0.0000 0.0000 0.0000 0.0000 0.0000 0.0000 0.0000 0.0000 alpha-Ketoglutarate_12345_13C_2 0.0000 0.0000 0.0000 0.0000 0.0000 0.0000 0.0000 0.0000 0.0000 0.0000 0.0000 0.0000 0.0000 0.0000 0.0000 0.0000 alpha-Ketoglutarate_12345_13C_3 0.0000 0.0000 29.5832 0.0000 0.0000 0.0000 0.0000 0.0000 0.0000 0.0000 0.0000 0.0000 0.0000 0.0000 0.0000 0.0000 alpha-Ketoglutarate_12345_13C_4 0.0000 0.0000 0.3796 0.0000 0.0000 0.0000 0.0000 0.0000 0.0000 0.0000 0.0000 0.0000 0.0000 0.0000 0.0000 0.0000 alpha-Ketoglutarate_12345_13C_5 0.0000 0.0000 3953.5164 0.0000 434.5178 0.0000 0.0000 0.0000 0.0000 0.0000 0.0000 0.0000 0.0000 0.0000 0.0000 0.0000 Aspartate_1234 29592.8259 17474.2317 9398.3528 18541.1619 8756.3015 20359.9893 17422.4964 15816.6190 28656.9980 21186.7341 13502.3550 18571.2894 25251.5001 13814.0355 22351.0978 20576.3498 Aspartate_1234_13C_1 174.6987 0.0000 640.9797 2193.7171 766.0561 2413.7347 1285.0032 2108.5841 0.0000 0.0000 1950.2143 6008.0164 4190.1066 3498.6963 6248.4231 4248.9807 Aspartate_1234_13C_2 0.0000 0.0000 1517.6518 2047.4518 1608.7041 1381.5293 1315.5481 1351.2706 0.0000 35.9563 3901.9791 5659.5430 6244.4469 4395.0843 6578.2891 5610.3444 Aspartate_1234_13C_3 71.4307 50.2060 1560.6206 264.6789 812.7786 0.0000 235.5518 106.5318 100.5012 0.0000 1497.1132 2118.8550 2982.6069 1226.0320 2364.0365 2655.5530 Aspartate_1234_13C_4 0.0000 20.6182 13382.5429 2458.9571 6363.0152 1926.4878 1607.3336 1301.7518 0.0000 0.0000 260.6253 336.9104 419.7247 447.4419 307.7502 496.4255 Fumarate_1234 0.0000 0.0000 7381.4407 1305.7072 3818.8015 359.6258 0.0000 0.0000 0.0000 0.0000 524.4448 790.1094 679.3422 302.8856 472.0050 1047.6665 Fumarate_1234_13C_1 0.0000 0.0000 340.6684 0.0000 11.6266 0.0000 0.0000 0.0000 0.0000 0.0000 122.1028 0.0000 0.0000 0.0000 0.0000 31.6463 Fumarate_1234_13C_2 0.0000 0.0000 291.8893 0.0000 622.1592 0.0000 0.0000 0.0000 0.0000 0.0000 0.0000 0.0000 485.1552 0.0000 0.0000 123.1158 Fumarate_1234_13C_3 0.0000 0.0000 0.0000 0.0000 0.0000 0.0000 0.0000 0.0000 0.0000 0.0000 0.0000 0.0000 0.0000 0.0000 0.0000 0.0000 Fumarate_1234_13C_4 0.0000 0.0000 3560.1586 0.0000 1151.4196 0.0000 0.0000 0.0000 0.0000 0.0000 0.0000 0.0000 0.0000 0.0000 0.0000 0.0000 Glutamate_12345 25599.7954 14794.9491 65384.4337 99292.7936 75767.3704 98276.7514 83825.1140 77320.7604 23398.9203 17391.5553 87727.5358 136388.5560 152256.7278 91158.5901 130685.2609 130167.9423 Glutamate_12345_13C_1 0.0000 0.0000 2479.8346 10843.5905 5487.8771 11611.7649 12762.6570 12110.0237 0.0000 0.0000 18439.7144 24947.7330 29903.6796 19072.8433 28722.3490 29265.5152 Glutamate_12345_13C_2 0.0000 77.7480 3276.2272 8501.4109 4305.0174 6728.6898 5882.9484 4576.4960 0.0000 27.0650 36618.9972 55695.1077 64273.3484 37352.8604 54965.1075 57534.2071 Glutamate_12345_13C_3 214.3459 23.2387 16588.6554 11097.4868 18874.3853 9290.9237 9376.2749 8623.8545 112.5054 0.0000 10880.4631 16814.1605 21994.6025 11862.4461 18980.4115 16012.7977 Glutamate_12345_13C_4 0.0000 0.0000 18255.4369 1012.6364 15165.6728 1069.2774 624.2308 794.5861 0.0000 0.0000 6125.3210 9165.3105 10900.2901 7462.1347 10221.9409 11143.1268 Glutamate_12345_13C_5 0.0000 0.0000 284935.4653 34046.4493 245798.8450 15574.6939 15933.5209 13067.6849 41.4132 0.0000 1947.8931 2361.8262 1357.8632 2012.1045 2326.9385 1891.5348 Glutamine_12345 0.0000 101306.7006 0.0000 0.0000 0.0000 0.0000 0.0000 0.0000 0.0000 135045.7008 12041.1686 14932.5114 21293.3600 10386.0907 13733.4740 19115.3033 Glutamine_12345_13C_1 0.0000 1705.7257 0.0000 0.0000 0.0000 0.0000 0.0000 0.0000 320.8698 558.5260 0.0000 647.7343 0.0000 128.1029 0.0000 0.0000 Glutamine_12345_13C_2 0.0000 0.0000 0.0000 0.0000 0.0000 0.0000 0.0000 0.0000 188.0370 0.0000 0.0000 0.0000 303.6519 0.0000 29.1594 0.0000 Glutamine_12345_13C_3 0.0000 0.0000 0.0000 0.0000 0.0000 0.0000 0.0000 0.0000 308.8884 0.0000 0.0000 0.0000 0.0000 0.0000 0.0000 0.0000 Glutamine_12345_13C_4 0.0000 4189.7327 0.0000 0.0000 0.0000 0.0000 0.0000 0.0000 627.7733 5584.8692 310.0349 0.0000 626.4156 0.0000 696.4377 646.4842 Lactate_123 4462.0446 2736.0241 191981.1755 35055.9455 140062.8571 15519.0921 15921.5653 15319.3651 4508.2463 3546.1406 22780.0986 26301.1198 41118.4212 26210.1578 32130.7661 31057.0447 Lactate_123_13C_1 36.8328 0.0000 0.0000 0.0000 0.0000 0.2262 0.0000 240.7125 0.0000 44.6009 264.0385 1338.1624 1862.3258 497.3326 863.0899 1499.6713 Lactate_123_13C_2 780.1054 571.6041 7146.9050 3860.7921 5697.0279 3714.3170 3248.3381 3665.1669 813.7714 505.4093 15147.2776 21479.8147 24527.4721 17684.7891 23558.4474 24683.8265 Lactate_123_13C_3 52.8954 68.3466 4612.5259 0.0000 2383.6020 116.6088 430.7699 0.0000 129.9999 20.1264 526.9400 745.4781 684.9722 420.0098 215.1580 1136.6636 Oxaloacetate_1234 2307.7200 1785.8590 10896.9402 35015.6587 25823.7422 35945.6804 35150.5173 34018.1929 2498.3206 1901.1957 19945.1481 36853.5901 37491.2754 23115.6800 26621.4090 31581.2402 Oxaloacetate_1234_13C_1 31.1232 37.2352 223.6551 0.0000 0.0000 477.3396 351.0738 47.4765 0.0000 73.0315 0.0000 0.0000 0.0000 0.0000 0.0000 212.9942 Oxaloacetate_1234_13C_2 21.5485 0.0000 0.0000 0.0000 0.0000 0.0000 0.0000 0.0000 0.0000 0.0000 42.7728 0.0000 0.0000 252.4424 0.0000 0.0000 Oxaloacetate_1234_13C_3 0.0000 0.0000 0.0000 0.0000 0.0000 0.0000 0.0000 0.0000 0.0000 0.0000 0.0000 43.7545 81.8673 0.0000 47.8062 5.0632 Oxaloacetate_1234_13C_4 0.0000 0.0000 0.0000 0.0000 0.0000 0.0000 0.0000 0.0000 0.0000 0.0000 0.0000 0.0000 0.0000 0.0000 0.0000 0.0000 Pyruvate_123 206.9248 122.6500 27400.9351 381.0424 7075.6535 0.0000 0.0000 0.0000 231.8081 165.1394 1469.4936 1678.8646 3054.4453 862.4860 1705.5571 1839.9192 Pyruvate_123_13C_1 163.8702 0.0000 989.7867 0.0000 475.2554 0.0000 0.0000 0.0000 0.0000 0.0000 221.5190 339.0851 493.4682 98.8319 627.0531 403.4013 Pyruvate_123_13C_2 0.0000 0.0000 169.7792 0.0000 78.9985 0.0000 0.0000 0.0000 0.0000 0.0000 869.6686 1419.0073 1668.1936 976.8516 1523.3026 1563.6397 Pyruvate_123_13C_3 0.0000 0.0000 2595.9289 0.0000 392.0355 0.0000 0.0000 0.0000 0.0000 0.0000 0.0000 100.8682 0.0000 0.0000 0.0000 0.0000 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 Alanine_123 5950 ALA_123 UM_Target_ID Alanine_123_13C_1 5950 ALA_123_13C_1 UM_Target_ID Alanine_123_13C_2 5950 ALA_123_13C_2 UM_Target_ID Alanine_123_13C_3 5950 ALA_123_13C_3 UM_Target_ID alpha-Ketoglutarate_12345 51 AKG_12345 UM_Target_ID alpha-Ketoglutarate_12345_13C_1 51 AKG_12345_13C_1 UM_Target_ID alpha-Ketoglutarate_12345_13C_2 51 AKG_12345_13C_2 UM_Target_ID alpha-Ketoglutarate_12345_13C_3 51 AKG_12345_13C_3 UM_Target_ID alpha-Ketoglutarate_12345_13C_4 51 AKG_12345_13C_4 UM_Target_ID alpha-Ketoglutarate_12345_13C_5 51 AKG_12345_13C_5 UM_Target_ID Aspartate_1234 424 ASP_1234 UM_Target_ID Aspartate_1234_13C_1 424 ASP_1234_13C_1 UM_Target_ID Aspartate_1234_13C_2 424 ASP_1234_13C_2 UM_Target_ID Aspartate_1234_13C_3 424 ASP_1234_13C_3 UM_Target_ID Aspartate_1234_13C_4 424 ASP_1234_13C_4 UM_Target_ID Fumarate_1234 444972 FUM_1234 UM_Target_ID Fumarate_1234_13C_1 444972 FUM_1234_13C_1 UM_Target_ID Fumarate_1234_13C_2 444972 FUM_1234_13C_2 UM_Target_ID Fumarate_1234_13C_3 444972 FUM_1234_13C_3 UM_Target_ID Fumarate_1234_13C_4 444972 FUM_1234_13C_4 UM_Target_ID Glutamate_12345 611 GLU_12345 UM_Target_ID Glutamate_12345_13C_1 611 GLU_12345_13C_1 UM_Target_ID Glutamate_12345_13C_2 611 GLU_12345_13C_2 UM_Target_ID Glutamate_12345_13C_3 611 GLU_12345_13C_3 UM_Target_ID Glutamate_12345_13C_4 611 GLU_12345_13C_4 UM_Target_ID Glutamate_12345_13C_5 611 GLU_12345_13C_5 UM_Target_ID Glutamine_12345 5961 GLN_12345 UM_Target_ID Glutamine_12345_13C_1 5961 GLN_12345_13C_1 UM_Target_ID Glutamine_12345_13C_2 5961 GLN_12345_13C_2 UM_Target_ID Glutamine_12345_13C_3 5961 GLN_12345_13C_3 UM_Target_ID Glutamine_12345_13C_4 5961 GLN_12345_13C_4 UM_Target_ID Lactate_123 612 LAC_123 UM_Target_ID Lactate_123_13C_1 612 LAC_123_13C_1 UM_Target_ID Lactate_123_13C_2 612 LAC_123_13C_2 UM_Target_ID Lactate_123_13C_3 612 LAC_123_13C_3 UM_Target_ID Oxaloacetate_1234 970 OAA_1234 UM_Target_ID Oxaloacetate_1234_13C_1 970 OAA_1234_13C_1 UM_Target_ID Oxaloacetate_1234_13C_2 970 OAA_1234_13C_2 UM_Target_ID Oxaloacetate_1234_13C_3 970 OAA_1234_13C_3 UM_Target_ID Oxaloacetate_1234_13C_4 970 OAA_1234_13C_4 UM_Target_ID Pyruvate_123 1060 PYR_123 UM_Target_ID Pyruvate_123_13C_1 1060 PYR_123_13C_1 UM_Target_ID Pyruvate_123_13C_2 1060 PYR_123_13C_2 UM_Target_ID Pyruvate_123_13C_3 1060 PYR_123_13C_3 UM_Target_ID METABOLITES_END #END