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Lysates were incubated on ice for 30min, sonicated for 10 seconds at 40% amplitude, and centrifuged at 14000 rcf for 10 minutes at 4°C. The supernatant was collected, and protein concentration was determined using Pierce BCA Protein Assay Kit (ThermoFisher Scientific) and a Molecular Devices SpectraMax iD3. A total of 30µg of protein was reduced with TCEP (ThermoFisher Scientific) to a concentration of 10mM and alkylated with IAA (Sigma) to a concentration of 30mM. Peptides underwent chloroform/methanol extraction (Fisher Chemical) and digested overnight at 47°C with sequencing grade modified porcine trypsin (Promega) in Triethylammonium bicarbonate (TEAB, Honeywell Fluka) at a protein:trypsin ratio of 50:1. The following day, samples were acidified to 0.5% formic acid/0.1% trifluoroacetic acid (TFA, Fisher Chemical). After plate equilibration with Buffer B, tryptic peptides were loaded onto a Sep-Pak C18 96-well plate (Waters) to be desalted using a Resolvex A200 (Tecan) automated method where samples underwent four washes with Buffer A. Peptides were eluted from the plate using 40% Buffer B. After equilibrating the plate with 70% Buffer B, samples were filtered using an AcroPrep Advance Filter Plate (Pall). Samples were lyophilized overnight (SpeedVac SPD210, ThermoFisher Scientific) and stored at -80°C. Buffer A = 0.1% formic acid, 0.5% acetonitrile in water, Buffer B = 80% acetonitrile, 20% water, 0.1% formic acid."],"repository":["Pride"],"quantification_method":[""],"modification":[""],"data_protocol":["Raw files acquired from abundance LC-MS analysis were analyzed with Spectronaut software (version 18.2) using the directDIA method with MS2 scans used for protein and peptide quantification. The UniProt murine proteome reference (Mouse_UP000000589_10090) containing 21,756 protein entries was used with protein and peptide-level false discovery rates (FDR) set to 0.01. Selected variable modifications included N-terminal acetylation and methionine oxidation while cysteine carbamidomethylation was set as a fixed modification. Trypsin was selected as the digestion enzyme with a maximum of two allowed missed cleavages and a minimum peptide length of seven amino acids. A total of 90,392 peptides and 7,520 protein groups were identified. Protein MS2 exclusive intensity values were quality-assessed using ProteiNorm99. Data normalization was performed with Variance Stabilizing Normalization (VSN)101, followed by statistical analysis using proteoDA102. Linear Models for Microarray Data (limma) combined with empirical Bayes (eBayes) smoothing were applied to the standard errors103. Proteins with a false discovery rate (FDR)-adjusted p-value of less than 0.05 were considered statistically significant.  Raw files acquired from SILAC LC-MS analysis were analyzed with Spectronaut software (version 18.2). Using the Pulsar search engine, a DIA-based spectral library was created using the “Generate Library from Pulsar/Search Archives” function. The UniProt murine proteome reference (Mouse_UP000000589_10090) containing 21,756 protein entries was used with protein and peptide-level false discovery rates (FDR) set to 0.01. Selected variable modifications included N-terminal acetylation and methionine oxidation while cysteine carbamidomethylation was set as a fixed modification. Trypsin was selected as the digestion enzyme with a maximum of two allowed missed cleavages and a minimum peptide length of seven amino acids. A total of 98,658 peptides and 7,834 protein groups were identified. A labeled spectral library was then generated, utilizing two label channels with Arg10 and Lys8 modifications added to the second channel. Raw files were searched using the labeled spectral library with MS2 scans used for protein and peptide quantification and half-life determination."],"omics_type":["Proteomics"],"labhead":["Ananda Goldrath"],"instrument_platform":[""],"submission_type":["COMPLETE"],"labhead_affiliation":["University of California San Diego"],"species":["Homo Sapiens (human)"],"submitter_mail":["bskoss@uams.edu"],"publication":["10.1016/J.CELL.2026.02.019"],"submitter_affiliation":["University of Arkansas for Medical Sciences"],"submitter_country":["United States"],"doi":["10.6019/PXD067417"],"additional_accession":[]},"is_claimable":false,"name":"Proteostasis sustains T cell differentiation potential and tumor-infiltrating lymphocyte function","description":"Tumors that contain tumor-infiltrating lymphocytes (TIL) with tissue-resident memory T cells (TRM) features are associated with better patient prognoses. While CD8+ T cells that target tumors often fail to restrain tumor growth due to progressive T cell exhaustion, pathogen-specific TRM survey tissues and sustain protective capacity for years. We identified protein homeostasis (proteostasis)-related E3 ubiquitin ligase genes shared by TRM and progenitor-exhausted TIL (TPEX), that are lost as TIL become terminally-exhausted (TEX). Sustained ligase expression led to increased TCF1+ T cell accumulation and improved anti-tumor function and less T cell exhaustion in chronic infection. Loss-of-function alleles in T cells challenged with cancer models impaired TIL accumulation and function and altered T cell differentiation in response to acute infection. Mass spectrometry revealed abundant expression of protein degradation pathways by TEX, yet they accumulated unfolded proteins. Ligase overexpression rescued unfolded protein accumulation in TEX, demonstrating their functional contribution to proteostasis. Ligase overexpression in combination with immune checkpoint blockade led to improved survival from murine melanoma, demonstrating that the relationship between proteostasis and T cell differentiation opens up new avenues for cancer immunotherapy.","dates":{"publication":"2026-04-30","submission":"2025-08-16"},"accession":"PXD067417","cross_references":{"TAXONOMY":["NEWT:330879","NEWT:377960","NEWT:1131","NEWT:2042546","NEWT:259447","NEWT:295546","NEWT:1334565","NEWT:625351","NEWT:258594","NCBITaxon:1280","NEWT:112503","NEWT:1129","NEWT:309807","NEWT:59753","NEWT:89184","NEWT:309800","NEWT:281395","NEWT:1211601","NEWT:876138","NEWT:44271","NEWT:193516","NEWT:1073882","NEWT:6819","NEWT:475174","NEWT:1117","NEWT:498257","NEWT:10036","NEWT:1590","NEWT:10034","NEWT:33114","NEWT:661410","NEWT:638632","NEWT:224326","NEWT:376619","NCBITaxon:79857","NEWT:1096976","NEWT:3765","NEWT:3767","NEWT:1589","NEWT:135622","NEWT:35786","NEWT:1580","NEWT:399784","NEWT:96731","NEWT:656064","NEWT:1582","NEWT:1584","NEWT:36630","NEWT:383379","NEWT:418106","NCBITaxon:12721","NEWT:10029","NEWT:913645","NEWT:641809","NEWT:1392488","NEWT:38815","NEWT:317447","NEWT:4688","NEWT:7719","NEWT:868565","NEWT:135674","NEWT:79329","NEWT:30069","NEWT:12637","NEWT:59729","NEWT:2164133","NEWT:295105","NEWT:108061","NEWT:60711","NEWT:224308","NEWT:3347","NEWT:160621","NEWT:212790","NEWT:110368","NEWT:1310161","NEWT:77133","NEWT:145481","NEWT:1310165","NEWT:29058","NCBITaxon:79824","NEWT:410658","NEWT:5516","NEWT:44688","NEWT:44689","NEWT:498211","NEWT:347256","NEWT:5518","NEWT:398007","NEWT:1527468","NEWT:498217","NEWT:498216","NEWT:11320","NEWT:246196","NEWT:246197","NEWT:145458","NEWT:1310173","NEWT:44685","NEWT:161934","NEWT:1148","NEWT:5508","NEWT:3329","NEWT:419481","NEWT:5507","NEWT:410661","NEWT:1140","NCBITaxon:2157","NEWT:1143","NEWT:1287689","NEWT:1094343","NEWT:1462472","NEWT:1336795","NEWT:644042","NEWT:150808","NEWT:1182590","NEWT:3712","NEWT:3711","NEWT:3714","NCBITaxon:37162","NEWT:2065263","NEWT:177437","NEWT:10418","NEWT:118698","NEWT:1616117","NEWT:118696","NEWT:34865","NEWT:52283","NEWT:284812","NEWT:8175","NEWT:43330","NEWT:1603293","NEWT:44664","NEWT:43346","NEWT:3702","NEWT:1245466","NEWT:3704","NEWT:244366","NEWT:1246791","NEWT:176597","NEWT:118694","NEWT:2850","NEWT:118691","NEWT:34871","NEWT:33548","NEWT:52299","NEWT:8165","NEWT:96794","NEWT:3708","NEWT:332648","NEWT:44670","NEWT:926571","NEWT:536231","NEWT:376219","NEWT:219813","NEWT:1510","NEWT:460519","NEWT:1515","NEWT:572307","NEWT:1432138","NEWT:1424507","NEWT:1194599","NEWT:272844","NEWT:1348799","NEWT:33995","NEWT:55784","NEWT:1303443","NEWT:485","NEWT:56636","NEWT:2499525","NEWT:2853422","NEWT:487","NEWT:1538553","NEWT:1679718","NEWT:480","NEWT:67767","NEWT:46835","NEWT:109757","NEWT:582580","NEWT:1185650","NEWT:216778","NEWT:1502","NEWT:128017","NEWT:376686","NEWT:237610","NEWT:95486","NEWT:1883446","NEWT:1233435","NEWT:118223","NEWT:11757","NEWT:987059","NEWT:109760","NEWT:29031","NEWT:317047","NEWT:491","NEWT:460523","NEWT:45954","NEWT:235443","NEWT:1757312","NEWT:85694","NEWT:109786","NEWT:108458","NEWT:5936","NEWT:320637","NEWT:1078283","NEWT:3750","NEWT:983964","NEWT:11706","NEWT:32644","NEWT:527796","NEWT:499175","NEWT:109779","NEWT:3745","NEWT:1715989","NCBITaxon:4751","NEWT:3747","NEWT:1116234","NEWT:1255228","NEWT:410289","NEWT:373153","NEWT:1640278","NEWT:472","NEWT:940825","NEWT:529507","NEWT:1071661","NEWT:28128","NEWT:470","NEWT:5911","NEWT:748693","NCBITaxon:50557","NEWT:39251","NEWT:29491","NEWT:101841","NEWT:446","NEWT:153481","NEWT:2014887","NEWT:33952","NEWT:445","NEWT:153009","NEWT:261756","NEWT:166313","NEWT:63366","NEWT:63367","NEWT:215402","NEWT:1547","NEWT:1290454","NEWT:906914","NEWT:28141","NEWT:9031","NEWT:27292","NEWT:1050722","NEWT:3728","NEWT:108931","NEWT:1293497","NEWT:1055524","NEWT:150475","NEWT:267872","NEWT:381666","NEWT:172269","NEWT:9534","NEWT:5180","NEWT:256737","NEWT:9541","NEWT:8694","NEWT:33936","NEWT:8692","NEWT:2903","NEWT:185579","NEWT:13076","NEWT:207340","NEWT:1006581","NEWT:8673","NEWT:33940","NEWT:550","NEWT:554","NEWT:451516","NEWT:552","NEWT:1325291","NEWT:42410","NEWT:36185","NEWT:1054211","NEWT:1225786","NEWT:771870","NEWT:51338","NEWT:575412","NEWT:28112","NEWT:49012","NEWT:6493","NEWT:10832","NEWT:6494","NEWT:6491","NEWT:507601","NEWT:520","NEWT:186441","NEWT:643680","NEWT:214092","NCBITaxon:6157","NEWT:13095","NEWT:162425","NEWT:104105","NEWT:216257","NEWT:9986","NEWT:8654","NEWT:8658","NEWT:1268063","NEWT:8657","NEWT:9983","NEWT:8655","NEWT:5147","NEWT:28108","NEWT:28104","NEWT:407821","NEWT:568703","NCBITaxon:2","NEWT:568708","NEWT:1613","NEWT:986","NEWT:52641","NEWT:28532","NEWT:353152","NEWT:298176","NEWT:353153","NEWT:227377","NEWT:1774284","NEWT:40674","NEWT:1194669","NEWT:443906","NEWT:13037","NEWT:519","NEWT:2510939","NEWT:7394","NEWT:6063","NEWT:1328388","NEWT:105487","NEWT:1548728","NEWT:667127","NEWT:9557","NEWT:377586","NEWT:300641","NEWT:39655","NEWT:9554","NEWT:475932","NEWT:256318","NEWT:206411","NCBITaxon:6191","NEWT:229533","NEWT:214053","NEWT:28550","NEWT:80863","NEWT:90675","NEWT:52639","NEWT:52638","NEWT:57075","NEWT:1666905","NEWT:8697","NEWT:861109","NEWT:8695","NEWT:884204","NEWT:1045775","NEWT:1123869","NEWT:9544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