{
"METABOLOMICS WORKBENCH":{"STUDY_ID":"ST002375","ANALYSIS_ID":"AN003870","VERSION":"1","CREATED_ON":"November 30, 2022, 2:23 am"},

"PROJECT":{"PROJECT_TITLE":"Metabolomics analysis of WT or GOT1 knockout CD8+ T cells cultured in serine-replete or serine-free media","PROJECT_SUMMARY":"WT or GOT1 knockout CD8+ T cells were activated with plate-bound anti-CD3 or soluble anti-CD28, in serine-replete or serine-free media for 24 hours. Intracellular metabolome were assessed by MS.","INSTITUTE":"Johns Hopkins University","LAST_NAME":"Xu","FIRST_NAME":"Wei","ADDRESS":"1650 Orleans Street, Baltimore, MD 21287, USA.","EMAIL":"wxu29@jhmi.edu","PHONE":"443-220-9936"},

"STUDY":{"STUDY_TITLE":"Metabolomics analysis of WT or GOT1 knockout CD8+ T cells cultured in serine-replete or serine-free media","STUDY_SUMMARY":"WT or GOT1 knockout CD8+ T cells were activated with plate-bound anti-CD3 or soluble anti-CD28, in serine-replete or serine-free media for 24 hours. Intracellular metabolome were assessed by MS.","INSTITUTE":"Johns Hopkins University","LAST_NAME":"Xu","FIRST_NAME":"Wei","ADDRESS":"1650 Orleans Street, Baltimore, MD 21287, USA.","EMAIL":"wxu29@jhmi.edu","PHONE":"443-220-9936"},

"SUBJECT":{"SUBJECT_TYPE":"Cultured cells","SUBJECT_SPECIES":"Mus musculus","TAXONOMY_ID":"10090"},
"SUBJECT_SAMPLE_FACTORS":[
{
"Subject ID":"-",
"Sample ID":"sample 1",
"Factors":{"Treatment":"WT_Serine-free"},
"Additional sample data":{"RAW_FILE_NAME":"sample 1.wiff"}
},
{
"Subject ID":"-",
"Sample ID":"sample 2",
"Factors":{"Treatment":"WT_Serine-free"},
"Additional sample data":{"RAW_FILE_NAME":"sample 2.wiff"}
},
{
"Subject ID":"-",
"Sample ID":"sample 3",
"Factors":{"Treatment":"WT_Serine-free"},
"Additional sample data":{"RAW_FILE_NAME":"sample 3.wiff"}
},
{
"Subject ID":"-",
"Sample ID":"sample 4",
"Factors":{"Treatment":"GOT1 KO_Serine-free"},
"Additional sample data":{"RAW_FILE_NAME":"sample 4.wiff"}
},
{
"Subject ID":"-",
"Sample ID":"sample 5",
"Factors":{"Treatment":"GOT1 KO_Serine-free"},
"Additional sample data":{"RAW_FILE_NAME":"sample 5.wiff"}
},
{
"Subject ID":"-",
"Sample ID":"sample 6",
"Factors":{"Treatment":"GOT1 KO_Serine-free"},
"Additional sample data":{"RAW_FILE_NAME":"sample 6.wiff"}
},
{
"Subject ID":"-",
"Sample ID":"sample 7",
"Factors":{"Treatment":"WT_Serine-replete"},
"Additional sample data":{"RAW_FILE_NAME":"sample 7.wiff"}
},
{
"Subject ID":"-",
"Sample ID":"sample 8",
"Factors":{"Treatment":"WT_Serine-replete"},
"Additional sample data":{"RAW_FILE_NAME":"sample 8.wiff"}
},
{
"Subject ID":"-",
"Sample ID":"sample 9",
"Factors":{"Treatment":"WT_Serine-replete"},
"Additional sample data":{"RAW_FILE_NAME":"sample 9.wiff"}
},
{
"Subject ID":"-",
"Sample ID":"sample 10",
"Factors":{"Treatment":"GOT1 KO_Serine-replete"},
"Additional sample data":{"RAW_FILE_NAME":"sample 10.wiff"}
},
{
"Subject ID":"-",
"Sample ID":"sample 11",
"Factors":{"Treatment":"GOT1 KO_Serine-replete"},
"Additional sample data":{"RAW_FILE_NAME":"sample 11.wiff"}
},
{
"Subject ID":"-",
"Sample ID":"sample 12",
"Factors":{"Treatment":"GOT1 KO_Serine-replete"},
"Additional sample data":{"RAW_FILE_NAME":"sample 12.wiff"}
}
],
"COLLECTION":{"COLLECTION_SUMMARY":"Cell pellets were spun down and washed twice with pre-warmed PBS. Metabolites were immediately extracted or stored at -80℃ until further extraction.","SAMPLE_TYPE":"Cultured cells"},

"TREATMENT":{"TREATMENT_SUMMARY":"CD8+ T cells were isolated from spleens and lymph nodes of WT or T cell conditional GOT1 knockout mice. Cells were then activated with plate-bound anti-CD3 and soluble anti-CD28 for 24 hours in normal media (serine-replete) or serine-free media."},

"SAMPLEPREP":{"SAMPLEPREP_SUMMARY":"Cell pellets were spun down and washed twice with pre-warmed PBS. Metabolites were immediately extracted by adding methanol:water (80:20, v/v) extraction solution, sonicated and stored at -80 °C for at least 2 hours to precipitate the proteins. Supernatant after centrifugation at 14,000xg for 10 minutes was dried under nitrogen gas. Metabolites were then reconstituted using ACN:water (50:50, v/v) overnight at 4 °C. Soluble metabolites after centrifugation at 14,000xg for 10 minutes were subjected to targeted metabolite analysis by liquid-chromatography tandem mass spectrometry (LC-MS/MS)."},

"CHROMATOGRAPHY":{"CHROMATOGRAPHY_TYPE":"HILIC","INSTRUMENT_NAME":"Shimadzu Prominence UFLC","COLUMN_NAME":"Waters XBridge BEH Amide, 2.1 x 150 mm, 2.5 μm"},

"ANALYSIS":{"ANALYSIS_TYPE":"MS"},

"MS":{"INSTRUMENT_NAME":"ABI Sciex 5500 QTrap","INSTRUMENT_TYPE":"QTRAP","MS_TYPE":"ESI","ION_MODE":"UNSPECIFIED","MS_COMMENTS":"The optimized MS parameters were: ESI voltage was +5,000 V in positive ion mode and -4,500 V in negative ion mode; dwell time was 3 ms per SRM transition and the total cycle time was 1.57 seconds. Peak integration for each targeted metabolite in SRM transition was processed with MultiQuant software (v2.1, AB Sciex). The preprocessed data with integrated peak areas were exported from MultiQuant and re-imported into Metaboanalyst software for further data analysis (statistical analysis, principal component analysis, generating heatmap, enrichment analysis, etc.)."},

"MS_METABOLITE_DATA":{
"Units":"AUC",

"Data":[{"Metabolite":"Urea","sample 1":"361186637.4","sample 2":"385710386.2","sample 3":"368004409","sample 4":"383221809.5","sample 5":"402052679.6","sample 6":"406081549.9","sample 7":"375261049.2","sample 8":"359648732","sample 9":"353854228.8","sample 10":"376644952.5","sample 11":"359287039","sample 12":"340816844.2"},{"Metabolite":"glycine","sample 1":"23951.01032","sample 2":"16946.89795","sample 3":"35226.00957","sample 4":"24022.21445","sample 5":"34757.80601","sample 6":"23670.63573","sample 7":"27686.06703","sample 8":"32237.85208","sample 9":"30869.91718","sample 10":"24772.62355","sample 11":"21046.84084","sample 12":"39292.56365"},{"Metabolite":"alanine","sample 1":"4440909.256","sample 2":"4191890.018","sample 3":"4495945.895","sample 4":"6814211.709","sample 5":"7233721.265","sample 6":"6692994.617","sample 7":"4299124.746","sample 8":"3799659.531","sample 9":"4181851.605","sample 10":"3030890.876","sample 11":"3352020.884","sample 12":"3342158.944"},{"Metabolite":"choline","sample 1":"144108032.2","sample 2":"137989276.7","sample 3":"153547813","sample 4":"109421369.5","sample 5":"111246734.9","sample 6":"113673541.8","sample 7":"183805513.7","sample 8":"164341987.5","sample 9":"180079740.7","sample 10":"188762772.7","sample 11":"197138431","sample 12":"197794924.9"},{"Metabolite":"4-aminobutyrate","sample 1":"102296.2752","sample 2":"129383.5201","sample 3":"157739.5611","sample 4":"106890.3899","sample 5":"115290.539","sample 6":"136217.6164","sample 7":"196100.7593","sample 8":"170918.497","sample 9":"177458.2906","sample 10":"178774.3692","sample 11":"136016.8836","sample 12":"154345.0055"},{"Metabolite":"dimethylglycine","sample 1":"13508146.41","sample 2":"13492082.24","sample 3":"15373078.63","sample 4":"10897087.09","sample 5":"10286429.96","sample 6":"11384700.88","sample 7":"16848636.59","sample 8":"16713975.22","sample 9":"16696846.41","sample 10":"19073330.72","sample 11":"19658922.72","sample 12":"17962132.27"},{"Metabolite":"serine","sample 1":"1669193.895","sample 2":"1572387.971","sample 3":"1855636.351","sample 4":"626355.1301","sample 5":"626664.6922","sample 6":"922746.0643","sample 7":"6864198.088","sample 8":"7506243.13","sample 9":"7737822.823","sample 10":"4444911.469","sample 11":"4735960.478","sample 12":"4437715.999"},{"Metabolite":"cytosine","sample 1":"248041.7126","sample 2":"210184.3611","sample 3":"352057.2812","sample 4":"296135.6839","sample 5":"281256.6652","sample 6":"251999.0363","sample 7":"134463.9142","sample 8":"306543.8975","sample 9":"346650.5311","sample 10":"204393.013","sample 11":"309828.7112","sample 12":"230109.6891"},{"Metabolite":"creatinine","sample 1":"1401277.933","sample 2":"1272665.661","sample 3":"1363786.662","sample 4":"1259045.594","sample 5":"1358154.918","sample 6":"1026950.01","sample 7":"1321481.888","sample 8":"1370322.089","sample 9":"1491832.52","sample 10":"1448319.224","sample 11":"1686012.624","sample 12":"1454247.906"},{"Metabolite":"proline","sample 1":"29895217.82","sample 2":"27280388.62","sample 3":"32842786.29","sample 4":"34140745.62","sample 5":"30585663.17","sample 6":"34397655.19","sample 7":"32571453.39","sample 8":"29163763.7","sample 9":"28304420.16","sample 10":"33419713.75","sample 11":"37369017.4","sample 12":"35787858.58"},{"Metabolite":"betaine","sample 1":"351476363.8","sample 2":"339241978.1","sample 3":"362427951.9","sample 4":"373769116.4","sample 5":"372412954.3","sample 6":"379884592.3","sample 7":"374073680.3","sample 8":"367647665.8","sample 9":"343766088.9","sample 10":"372918978.1","sample 11":"415050589.2","sample 12":"391203720.1"},{"Metabolite":"valine","sample 1":"156734.7549","sample 2":"187778.6328","sample 3":"144479.8447","sample 4":"181299.3555","sample 5":"175510.9911","sample 6":"138229.0726","sample 7":"187444.178","sample 8":"164084.7722","sample 9":"256780.8104","sample 10":"151620.1655","sample 11":"214709.6708","sample 12":"137219.9643"},{"Metabolite":"threonine","sample 1":"3719301.855","sample 2":"4044814.621","sample 3":"4113574.588","sample 4":"3917828.635","sample 5":"4000396.8","sample 6":"3646903.988","sample 7":"4253113.755","sample 8":"3992173.965","sample 9":"4019626.172","sample 10":"3955034.205","sample 11":"4016554.442","sample 12":"4109698.829"},{"Metabolite":"creatine","sample 1":"6029572.738","sample 2":"5502739.756","sample 3":"5333512.893","sample 4":"5898898.589","sample 5":"6780251.583","sample 6":"5755141.176","sample 7":"7710445.738","sample 8":"6942817.502","sample 9":"7372601.86","sample 10":"7347348.453","sample 11":"8122219.212","sample 12":"6673600.172"},{"Metabolite":"nicotinamide","sample 1":"1492881.543","sample 2":"1505519.587","sample 3":"1895394.968","sample 4":"1489505.184","sample 5":"1859710.144","sample 6":"1756841.614","sample 7":"1829889.827","sample 8":"2055419.149","sample 9":"2218857.578","sample 10":"2757514.259","sample 11":"3165926.467","sample 12":"2900129.93"},{"Metabolite":"thymine","sample 1":"151028.9023","sample 2":"158336.0165","sample 3":"158160.4818","sample 4":"212379.002","sample 5":"178373.8789","sample 6":"172411.901","sample 7":"208514.2655","sample 8":"231238.0027","sample 9":"196281.9997","sample 10":"196213.6284","sample 11":"160992.5558","sample 12":"170822.8251"},{"Metabolite":"DL-Pipecolic acid","sample 1":"97764138.03","sample 2":"95324707.99","sample 3":"97123194.06","sample 4":"65807644.37","sample 5":"70258068","sample 6":"63015947.79","sample 7":"107889846.2","sample 8":"107122928.5","sample 9":"106642027.8","sample 10":"91586098.71","sample 11":"98716448.1","sample 12":"90841610.32"},{"Metabolite":"hydroxyproline","sample 1":"8992064.416","sample 2":"9040593.384","sample 3":"10279073.09","sample 4":"8102229.536","sample 5":"7860503.629","sample 6":"7973069.129","sample 7":"11813176.9","sample 8":"10542865.89","sample 9":"10702085.24","sample 10":"10584035.4","sample 11":"10146983.5","sample 12":"10706838.62"},{"Metabolite":"leucine-isoleucine","sample 1":"32543689.72","sample 2":"32521983.79","sample 3":"33862502.52","sample 4":"29678570.36","sample 5":"33604718.01","sample 6":"24342666.88","sample 7":"35628041.73","sample 8":"33652548.7","sample 9":"43103466.7","sample 10":"36293610.93","sample 11":"48253072.14","sample 12":"33965744.32"},{"Metabolite":"ornithine","sample 1":"3199178.104","sample 2":"4757736.427","sample 3":"4410784.295","sample 4":"4855384.056","sample 5":"5010041.257","sample 6":"4107389.528","sample 7":"4222228.052","sample 8":"4703223.977","sample 9":"6805634.691","sample 10":"6179494.956","sample 11":"6813691.259","sample 12":"6313992.196"},{"Metabolite":"asparagine","sample 1":"4476354.02","sample 2":"3799316.645","sample 3":"4182097.127","sample 4":"3612185.51","sample 5":"4035112.539","sample 6":"3827064.581","sample 7":"5088326.286","sample 8":"3912793.989","sample 9":"4318058.72","sample 10":"4597214.607","sample 11":"4598288.487","sample 12":"4342392.955"},{"Metabolite":"aspartate","sample 1":"29308881.31","sample 2":"25362501.52","sample 3":"34067146.81","sample 4":"69725841.72","sample 5":"63906731.36","sample 6":"70785718.22","sample 7":"44650857.44","sample 8":"37702903.17","sample 9":"38983650.03","sample 10":"91179340.74","sample 11":"91083549.21","sample 12":"86478245.16"},{"Metabolite":"adenine","sample 1":"410081.0073","sample 2":"565819.5507","sample 3":"541311.6899","sample 4":"331188.6148","sample 5":"392211.4509","sample 6":"394172.7816","sample 7":"855198.7289","sample 8":"799347.6598","sample 9":"923304.0871","sample 10":"774722.2162","sample 11":"1362407.918","sample 12":"906041.5979"},{"Metabolite":"homocysteine","sample 1":"128736.6502","sample 2":"93944.35984","sample 3":"159444.8127","sample 4":"246874.0795","sample 5":"273535.7062","sample 6":"261792.8197","sample 7":"185131.0206","sample 8":"161012.8628","sample 9":"115161.2867","sample 10":"353394.3564","sample 11":"336222.0828","sample 12":"404641.7513"},{"Metabolite":"lysine","sample 1":"75812.61822","sample 2":"108614.5967","sample 3":"156616.8805","sample 4":"127724.0523","sample 5":"137141.9095","sample 6":"123532.2395","sample 7":"108737.6606","sample 8":"129996.304","sample 9":"170258.2928","sample 10":"105893.4948","sample 11":"164889.9646","sample 12":"108207.6996"},{"Metabolite":"glutamine","sample 1":"58387287.82","sample 2":"59785628.46","sample 3":"57730271.02","sample 4":"46240591.37","sample 5":"48599219.24","sample 6":"42047507.46","sample 7":"59485346.49","sample 8":"60754026.51","sample 9":"68473819.85","sample 10":"56215726.91","sample 11":"56489283.71","sample 12":"58572497.4"},{"Metabolite":"O-acetyl-L-serine","sample 1":"149245.1837","sample 2":"125703.8585","sample 3":"180896.5162","sample 4":"134777.0728","sample 5":"120016.0002","sample 6":"132389.7623","sample 7":"187819.1529","sample 8":"189849.0034","sample 9":"154869.2016","sample 10":"189523.7655","sample 11":"214002.1721","sample 12":"161866.3993"},{"Metabolite":"glutamate","sample 1":"97375159.7","sample 2":"94948946.45","sample 3":"97620422.68","sample 4":"57030497.1","sample 5":"63498622.71","sample 6":"60113013.7","sample 7":"123799050.7","sample 8":"110700734.9","sample 9":"108604894.9","sample 10":"93424228.46","sample 11":"88293344.67","sample 12":"91407209.69"},{"Metabolite":"methionine","sample 1":"739887.0599","sample 2":"848605.6944","sample 3":"826274.1339","sample 4":"721637.2726","sample 5":"880491.3703","sample 6":"623717.3817","sample 7":"773668.0869","sample 8":"820095.8104","sample 9":"950561.272","sample 10":"756565.5295","sample 11":"987216.3479","sample 12":"702682.2229"},{"Metabolite":"histidine","sample 1":"1673759.787","sample 2":"1573213.399","sample 3":"1711308.079","sample 4":"1583198.613","sample 5":"1703579.201","sample 6":"1360855.686","sample 7":"1764095.82","sample 8":"1762251.385","sample 9":"2114471.987","sample 10":"1755405.478","sample 11":"2328294.648","sample 12":"2122359.294"},{"Metabolite":"2-Aminooctanoic acid","sample 1":"541006.1699","sample 2":"488378.3646","sample 3":"512224.4658","sample 4":"806900.6984","sample 5":"864436.1959","sample 6":"826630.7973","sample 7":"543642.1833","sample 8":"463401.6777","sample 9":"551634.8368","sample 10":"896257.0597","sample 11":"864670.0129","sample 12":"861173.9257"},{"Metabolite":"carnitine","sample 1":"19000607.14","sample 2":"17647733.82","sample 3":"19888750.57","sample 4":"20784629.81","sample 5":"20510029.89","sample 6":"21194780.66","sample 7":"21029298.96","sample 8":"22836104.47","sample 9":"18729646.98","sample 10":"21310456.3","sample 11":"24311371.06","sample 12":"23506917.19"},{"Metabolite":"Methionine sulfoxide","sample 1":"161812.9912","sample 2":"181630.5852","sample 3":"210968.5665","sample 4":"194537.401","sample 5":"193861.3895","sample 6":"171127.0009","sample 7":"191155.2529","sample 8":"207665.5218","sample 9":"240484.017","sample 10":"237948.872","sample 11":"258636.1813","sample 12":"214986.2298"},{"Metabolite":"phenylalanine","sample 1":"3726514.305","sample 2":"3689419.218","sample 3":"3816146.591","sample 4":"3557598.54","sample 5":"3962319.383","sample 6":"2801005.232","sample 7":"3384399.256","sample 8":"3833347.974","sample 9":"4789354.708","sample 10":"3631022.251","sample 11":"4580573.883","sample 12":"3474898.61"},{"Metabolite":"1-Methyl-Histidine","sample 1":"968389.1822","sample 2":"1259761.788","sample 3":"1555104.257","sample 4":"1364875.653","sample 5":"1672002.77","sample 6":"1340697.389","sample 7":"1523912.336","sample 8":"1891956.159","sample 9":"2156518.704","sample 10":"2101840.189","sample 11":"2543399.815","sample 12":"2081332.064"},{"Metabolite":"arginine","sample 1":"1409788.158","sample 2":"2224015.646","sample 3":"2307370.2","sample 4":"2487334.896","sample 5":"2839654.122","sample 6":"2208720.295","sample 7":"2479294.759","sample 8":"2506622.08","sample 9":"2922038.69","sample 10":"2471933.337","sample 11":"2845954.76","sample 12":"1978371.826"},{"Metabolite":"citrulline","sample 1":"165177.7457","sample 2":"130617.0093","sample 3":"155021.0039","sample 4":"176889.1509","sample 5":"160705.1897","sample 6":"158771.6572","sample 7":"158789.7937","sample 8":"134962.1811","sample 9":"191596.1245","sample 10":"198512.5564","sample 11":"205990.1179","sample 12":"145971.6383"},{"Metabolite":"tyrosine","sample 1":"895206.3791","sample 2":"937362.5516","sample 3":"1041683.769","sample 4":"876272.946","sample 5":"1127262.21","sample 6":"818161.5777","sample 7":"903094.2384","sample 8":"904626.2639","sample 9":"1134369.322","sample 10":"983128.9883","sample 11":"1249202.018","sample 12":"855185.9974"},{"Metabolite":"Phosphorylcholine","sample 1":"54500457.52","sample 2":"51501760.68","sample 3":"54524401.08","sample 4":"18219706.03","sample 5":"15249313.79","sample 6":"19276170.56","sample 7":"62934067.46","sample 8":"59655561.97","sample 9":"57851741.06","sample 10":"42905407.04","sample 11":"41895819.77","sample 12":"39726027.56"},{"Metabolite":"3-phospho-serine","sample 1":"141725.8075","sample 2":"99237.86146","sample 3":"117247.3624","sample 4":"25100.02796","sample 5":"13861.09037","sample 6":"19045.9607","sample 7":"1171690.151","sample 8":"931638.1975","sample 9":"1152843.881","sample 10":"53067.00672","sample 11":"45325.5525","sample 12":"44671.60657"},{"Metabolite":"N6-Acetyl-L-lysine","sample 1":"393925.7586","sample 2":"586254.2539","sample 3":"521647.9013","sample 4":"524350.0038","sample 5":"553345.8266","sample 6":"451026.249","sample 7":"549118.9823","sample 8":"592561.4265","sample 9":"608870.0951","sample 10":"512393.203","sample 11":"658619.3636","sample 12":"641253.5614"},{"Metabolite":"Acetyllysine","sample 1":"280664.0559","sample 2":"480982.1604","sample 3":"403437.1925","sample 4":"458317.0537","sample 5":"431971.4777","sample 6":"315217.267","sample 7":"410436.9744","sample 8":"420296.3459","sample 9":"538830.3556","sample 10":"424021.3921","sample 11":"588595.5895","sample 12":"411599.0158"},{"Metabolite":"N-acetyl-glutamine","sample 1":"173507.2796","sample 2":"168295.2364","sample 3":"164993.1951","sample 4":"150717.4604","sample 5":"174407.7738","sample 6":"129541.5856","sample 7":"192182.9605","sample 8":"245258.5966","sample 9":"197411.3194","sample 10":"198657.9005","sample 11":"268767.0857","sample 12":"185842.0636"},{"Metabolite":"N-acetyl-glutamate","sample 1":"69838.81393","sample 2":"92130.26499","sample 3":"136466.2059","sample 4":"3120392.306","sample 5":"2577860.924","sample 6":"1883611.291","sample 7":"1771832.313","sample 8":"1858163.996","sample 9":"2080314.242","sample 10":"1819417.035","sample 11":"1942398.781","sample 12":"2335336.386"},{"Metabolite":"Ng,NG-dimethyl-L-arginine","sample 1":"209970.3129","sample 2":"207317.0244","sample 3":"250373.8418","sample 4":"272360.5998","sample 5":"244329.9308","sample 6":"201912.4842","sample 7":"264771.7773","sample 8":"265873.8852","sample 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