#METABOLOMICS WORKBENCH amitch_20150922_9378651_mwtab.txt DATATRACK_ID:383 STUDY_ID:ST000249 ANALYSIS_ID:AN000393 PROJECT_ID:PR000201 VERSION 1 CREATED_ON 2016-09-17 #PROJECT PR:PROJECT_TITLE Myc-directed suppression of autophagy provides therapeutic vulnerabilities PR:PROJECT_TITLE amino acid homeostasis PR:PROJECT_TYPE Targeted LCMS Metabolomics PR:PROJECT_SUMMARY Hypothesis:MYC is suppressing the autophagy pathway which leads to impaired PR:PROJECT_SUMMARY of proteins and reduced free AA in the cells. PR:INSTITUTE Moffitt Cancer Center PR:DEPARTMENT Tumor Biology PR:LABORATORY John Cleveland PR:LAST_NAME Cleveland PR:FIRST_NAME John PR:ADDRESS 12902 Magnolia Dr PR:EMAIL Tampa, FL 33612 PR:PHONE 813 745 3888 PR:FUNDING_SOURCE NIH #STUDY ST:STUDY_TITLE Measurement of free amino acid (AA) in response to MYC ST:STUDY_TYPE treatment ST:STUDY_SUMMARY The proposed study will provide us information how the different AAs are ST:STUDY_SUMMARY by MYC. MYC is known to cause increased levels of glutamin due to import and ST:STUDY_SUMMARY but we hypothesize that the overall pool of free AA in the cells is reduced, ST:STUDY_SUMMARY when cells are starved. ST:INSTITUTE University of Florida ST:DEPARTMENT SECIM ST:LAST_NAME Schaub ST:FIRST_NAME Franz ST:ADDRESS - ST:EMAIL Franz.Schaub@moffitt.org ST:PHONE - ST:NUM_GROUPS 2 ST:TOTAL_SUBJECTS 6 #SUBJECT SU:SUBJECT_TYPE Human SU:SUBJECT_SPECIES Homo sapiens SU:TAXONOMY_ID 9606 SU:SPECIES_GROUP Human #SUBJECT_SAMPLE_FACTORS: SUBJECT(optional)[tab]SAMPLE[tab]FACTORS(NAME:VALUE pairs separated by |)[tab]Additional sample data SUBJECT_SAMPLE_FACTORS - 48hr_TET_1 Treatment:48 hour Tet treatment sample type=Cell pellet from P493-6 B Cells SUBJECT_SAMPLE_FACTORS - 48hr_TET_2 Treatment:48 hour Tet treatment sample type=Cell pellet from P493-6 B Cells SUBJECT_SAMPLE_FACTORS - 48hr_TET_3 Treatment:48 hour Tet treatment sample type=Cell pellet from P493-6 B Cells SUBJECT_SAMPLE_FACTORS - 48hr_NoTET_1 Treatment:No Tet treatment sample type=Cell pellet from P493-6 B Cells SUBJECT_SAMPLE_FACTORS - 48hr_NoTET_2 Treatment:No Tet treatment sample type=Cell pellet from P493-6 B Cells SUBJECT_SAMPLE_FACTORS - 48hr_NoTET_3 Treatment:No Tet treatment sample type=Cell pellet from P493-6 B Cells #COLLECTION CO:COLLECTION_SUMMARY Cells were counted, washed with cold PBS and then flash-frozen in liquid N2 CO:SAMPLE_TYPE Cells CO:VOLUMEORAMOUNT_COLLECTED 1.00E7 #TREATMENT TR:TREATMENT_SUMMARY Cells were treated with TET for 48 hrs (TET samples) and then washed and TR:TREATMENT_SUMMARY without TET for 48hrs (noTET samples) #SAMPLEPREP SP:SAMPLEPREP_SUMMARY Amino acid SP:SAMPLEPREP_PROTOCOL_FILENAME SB_AA_Assay.pdf #CHROMATOGRAPHY CH:CHROMATOGRAPHY_SUMMARY Amino Acid CH:CHROMATOGRAPHY_TYPE Reversed phase MS:INSTRUMENT_NAME Agilent 6490 QQQ CH:COLUMN_NAME Waters AccQ Tag (100 x 2mm, 1.7um) CH:METHODS_FILENAME SB_AA_Assay.pdf #ANALYSIS AN:ANALYSIS_TYPE MS AN:LABORATORY_NAME Sanford Burnham AN:DETECTOR_TYPE Triple Quadrupole #MS MS:INSTRUMENT_NAME Agilent 6490 QQQ MS:INSTRUMENT_TYPE Triple quadrupole MS:MS_TYPE ESI MS:ION_MODE POSITIVE MS:MS_COMMENTS Amino Acids #MS_METABOLITE_DATA MS_METABOLITE_DATA:UNITS nmol/million cells MS_METABOLITE_DATA_START Samples 48hr_TET_1 48hr_TET_2 48hr_TET_3 48hr_NoTET_1 48hr_NoTET_2 48hr_NoTET_3 Factors Treatment:48 hour Tet treatment Treatment:48 hour Tet treatment Treatment:48 hour Tet treatment Treatment:No Tet treatment Treatment:No Tet treatment Treatment:No Tet treatment Alanine 0.3090 0.2532 0.3071 0.3312 0.4396 0.4614 Arginine 0.1119 0.0880 0.0752 0.1316 0.1519 0.1505 Asparagine 0.3839 0.3703 0.3897 0.6176 0.7081 0.7002 Aspartic Acid 1.8494 1.6285 1.6821 1.3932 1.7418 1.6675 Citrulline Glutamic Acid 9.7901 8.9728 9.5972 9.8469 11.9768 10.2894 Glutamine 2.3819 2.0457 2.1411 2.5200 3.3155 3.4919 Glycine 1.4412 1.2484 1.1886 0.7270 1.0621 0.9695 Histidine 0.0695 0.0579 0.0615 0.0496 0.0784 0.0662 Isoleucine 0.1405 0.1207 0.1479 0.1329 0.1471 0.1665 Leucine 0.1531 0.1285 0.1525 0.1254 0.1441 0.1486 Lysine Methionine 0.0790 0.0780 0.0770 0.0695 0.0801 0.0772 Ornithine Phenylalanine 0.0557 0.0480 0.0552 0.0466 0.0598 0.0532 Proline 0.4964 0.4306 0.4714 1.0962 1.3621 1.4309 Serine 0.1952 0.1563 0.1813 0.1547 0.2017 0.2162 Threonine 0.1525 0.1243 0.1413 0.1462 0.2107 0.1861 Tryptophan 0.0120 0.0098 0.0129 0.0092 0.0127 0.0124 Tyrosine 0.0672 0.0652 0.0697 0.0670 0.0753 0.0756 Valine 0.0840 0.0796 0.0787 0.0536 0.0692 0.0650 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 260.17, 171 4.16 UFLORIDA_MS_RI 5950 C00041 UFLORIDA_FEATURE Arginine 345.21, 170.9 2.68 UFLORIDA_MS_RI 6322 C00062 UFLORIDA_FEATURE Asparagine 303.13, 170.9 2.25 UFLORIDA_MS_RI 6267 C00152 UFLORIDA_FEATURE Aspartic Acid 304.11, 171 2.99 UFLORIDA_MS_RI 5960 C00049 UFLORIDA_FEATURE Citrulline 346.21, 171 3.33 UFLORIDA_MS_RI 9750 C00327 UFLORIDA_FEATURE Glutamic Acid 318.11, 171.1 3.35 UFLORIDA_MS_RI 33032 UFLORIDA_FEATURE Glutamine 317.21, 171 2.65 UFLORIDA_MS_RI 5961 C00064 UFLORIDA_FEATURE Glycine 246.08, 170.9 2.81 UFLORIDA_MS_RI 750 C00037 UFLORIDA_FEATURE Histidine 326.21, 110 2.07 UFLORIDA_MS_RI 6274 C00135 UFLORIDA_FEATURE Isoleucine 302.21, 170.9 8.15 UFLORIDA_MS_RI 6306 C00407 UFLORIDA_FEATURE Leucine 302.21, 170.9 8.26 UFLORIDA_MS_RI 6106 C00123 UFLORIDA_FEATURE Lysine 487.21, 170.9 6.66 UFLORIDA_MS_RI 5962 C00047 UFLORIDA_FEATURE Methionine 320.21, 170.9 6.87 UFLORIDA_MS_RI 6137 C00073 UFLORIDA_FEATURE Ornithine 473.22, 170.9 5.93 UFLORIDA_MS_RI 6262 C00077 UFLORIDA_FEATURE Phenylalanine 336.21, 170.9 8.40 UFLORIDA_MS_RI 6140 C00079 UFLORIDA_FEATURE Proline 286.16, 171 4.84 UFLORIDA_MS_RI 145742 C00148 UFLORIDA_FEATURE Serine 276.11, 170.9 2.60 UFLORIDA_MS_RI 5951 C00065 UFLORIDA_FEATURE Threonine 290.11, 170.9 3.74 UFLORIDA_MS_RI 6288 C00188 UFLORIDA_FEATURE Tryptophan 375.21, 171 8.54 UFLORIDA_MS_RI 6305 C00078 UFLORIDA_FEATURE Tyrosine 352.21, 170.9 6.66 UFLORIDA_MS_RI 6057 C00082 UFLORIDA_FEATURE Valine 288.21, 171 7.09 UFLORIDA_MS_RI 6287 C00183 UFLORIDA_FEATURE METABOLITES_END #END