{"database":"Pride","file_versions":[{"headers":{"Content-Type":["application/json"]},"body":{"files":{"Xlsx":["ftp://ftp.pride.ebi.ac.uk/pride/data/archive/2026/05/PXD045820/Results.xlsx"],"Txt":["ftp://ftp.pride.ebi.ac.uk/pride/data/archive/2026/05/PXD045820/TPP_SDRF.txt"],"Mzml":["ftp://ftp.pride.ebi.ac.uk/pride/data/archive/2026/05/PXD045820/Eclipse_02766.mzML","ftp://ftp.pride.ebi.ac.uk/pride/data/archive/2026/05/PXD045820/Eclipse_02765.mzML"],"Raw":["ftp://ftp.pride.ebi.ac.uk/pride/data/archive/2026/05/PXD045820/Eclipse_02766.raw","ftp://ftp.pride.ebi.ac.uk/pride/data/archive/2026/05/PXD045820/Eclipse_02765.raw"],"Pepxml":["ftp://ftp.pride.ebi.ac.uk/pride/data/archive/2026/05/PXD045820/Eclipse_02765.pepXML","ftp://ftp.pride.ebi.ac.uk/pride/data/archive/2026/05/PXD045820/Eclipse_02766.pepXML"]},"type":"primary"},"statusCode":"OK","statusCodeValue":200}],"scores":null,"additional":{"labhead_mail":["milos.filipovic@isas.de"],"submitter":["Thibaut Vignane"],"technology_type":["Mass Spectrometry","Shotgun proteomics"],"software":[""],"submitter_keywords":[""],"full_dataset_link":["https://www.ebi.ac.uk/pride/archive/projects/PXD045820"],"tissue":["Embryonic Fibroblast","Cell Culture"],"sample_protocol":["90 % confluency 10 mm dish of WT and CSE-/- MEFs were trypsinized and washed three times in cold serum-free DMEM. After the last centrifugation, cells were resuspended in 1.2 ml of serum-free mediaDMEM supplemented with 10 % FCS and treated for 60 min with 500 µMm H2O2 on ice. After 60 min, cells were aliquoted and centrifuged. Cell pellets were resuspended in 20 µl serum-free media and heated at 25, 47, or 57 °C for 3 min. Cells were brought back to room temperature for 3 min and lysed using freeze-thaw cycles in liquid nitrogen and thawing on ice. Once lyszed, a final concentration of 50 mM ABC was added to samples, and extracted proteins were subject to 100,000 x g ultracentrifugation for 20 min at 4 °C to obtain soluble fractions for downstream analysis. Protein concentration was then measured, where protein amounts from 25 °C-exposed cells were used for sample re-adjustment across all temperatures. Proteins were subsequently denatured using 3M Guanidine-HCl (Gu-HCl) for reduction and alkylation steps with 500 µM DTT and 5 mM IAM for 30 min at 37 °C, respectively. ABC (50 mM) was added to reach a concentration of 0.3 M Gu-HCl for trypsin digestion using a 1:20 trypsin to sample ratio. Peptides were desalted using Supel™-Select HLB SPE Tube (Sigma, 54181-U) following the manufacturer’s protocol. Eluates were evaporated to dryness and resuspended in 100 mM triethylammonium bicarbonate (TEAB) for TMT labeling. Samples were labeled using the TMTpro™ 16plex Label Reagent Set (Thermo Fisher Scientific, #A44521). Samples were pulled down together (1:1, volume : volume), desalted, and dried as previously described. Peptides were dissolved in 0.1 % TFA, and digestion quality control was performed using an Ultimate 3000 Nano Ultra High-Pressure Chromatography (UPLC) system with a PepSwift Monolithic® Trap 200 µm * 5 mm (Thermo Fisher Scientific). Peptides were analyzed by high-resolution LC-MS/MS using an Ultimate 3000 Nano Ultra High-Pressure Chromatography (UPLC) system (Thermo Fisher Scientific) coupled with an Orbitrap Eclipse™ Tribrid™ Mass Spectrometer via an EASY-spray (Thermo Fisher Scientific). Peptide separation was carried out with an Acclaim™ PepMap™ 100 C18 column (Thermo Fisher Scientific) using a 90min gradient from 3 to 35 % of B (84 % Acetonitrile, 0.1 % Formic Acid) at a flow rate of 250 nL/min. The Orbitrap Eclipse™ was operated in a DDA mode, and MS1 survey scans were acquired from m/z 300 to 1,500 at a resolution of 120,000 using the Orbitrap mode. MS2 scans were carried out for 3 seconds with high-energy collision-induced dissociation (HCD) at 32 % using the Orbitrap mode at a resolution of 50,000."],"repository":["Pride"],"modification":[""],"quantification_method":[""],"data_protocol":["Data were evaluated using FragPipe(10) software using 0 ppm for precursor mass tolerance, 0.02 Da for fragment mass tolerance, specific tryptic digest, and a maximum of 3 missed cleavages.Carbamidomethylation(+57.021464 Da), N-term acetylation (+42.010565 Da), and methionine oxidation (+15.994915 Da) were added as variable modifications. TMTpro (+304.207146 Da) was added as fixe modification on K and peptide N-term, peptide-spectrum match (PSM) and proteins were filtered at FDR 1 %. TMT quantification was performed at the MS2 level."],"omics_type":["Proteomics"],"labhead":["Milos Filipovic"],"instrument_platform":[""],"labhead_affiliation":["SULFGAGING group, Leibniz-Institut für Analytische Wissenschaften – ISAS – e.V., Germany"],"species":["Mus Musculus (mouse)"],"submission_type":["PARTIAL"],"publication":["Not available"],"submitter_mail":["thibaut.vignane@outlook.com"],"submitter_affiliation":["Leibniz-Institut für Analytische Wissenschaften - ISAS,e.V, Dortmund, Germany"],"submitter_country":["Germany"],"additional_accession":[]},"is_claimable":false,"name":"Thermal proteome profilling in WT and CSE-/- MEF cells upon H2O2 treatment","description":"Cellular homeostasis relies on precise regulation through chemical processes, such as protein posttranslational modifications and physical processes, such as biomolecular condensation. Aging disrupts this balance, increasing susceptibility to diseases and death. However, the mechanisms behind age-related pathogenesis remain unclear. Using chemoproteomic methods to dissect different cysteine posttranslational modifications, we found that age-related changes in cysteine redox homeostasis impact proteins ability to form biomolecular condensates. Age-related increase in sulfenylation and sulfonylation promotes phase separation and through conformational change increases proteins propensity to aggregate. Causing condensates dissolution, protein persulfidation, a protective thiol modification, prevents this. Reduced persulfidation and increased sulfonylation in aging drive abnormal phase separation, leading to shorter life and spontaneous protein aggregation in CSE-/- mice.  In this study, we assessed the stability of proteins in WT and CSE-/- Mouse embryonic fibroblast mice upon H2O2 treatment.","dates":{"publication":"2026-05-23","submission":"2023-09-29"},"accession":"PXD045820","cross_references":{"TAXONOMY":["NEWT:330879","NEWT:377960","NEWT:2042546","NEWT:259447","NEWT:295546","NCBITaxon:2719036","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:475174","NEWT:1117","NEWT:498257","NEWT:10036","NEWT:1590","NEWT:10034","NEWT:661410","NEWT:638632","NEWT:224326","NEWT:376619","NCBITaxon:79857","NEWT:1096976","NEWT:1589","NEWT:135622","NEWT:35786","NEWT:1580","NEWT:399784","NEWT:96731","NEWT:383379","NEWT:418106","NEWT:10029","NEWT:913645","NEWT:641809","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: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: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:1182590","NEWT:3712","NEWT:3711","NEWT:270643","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:244366","NEWT:1246791","NEWT:118694","NEWT:2850","NEWT:118691","NEWT:34871","NEWT:33548","NEWT:52299","NEWT:96794","NEWT:3708","NEWT:332648","NEWT:44670","NEWT:536231","NEWT:376219","NEWT:219813","NEWT:1510","NEWT:460519","NEWT:1515","NEWT:572307","NEWT:1432138","NEWT:1424507","NEWT:1194599","NEWT:272844","NEWT:1303443","NEWT:9483","NEWT:485","NEWT:56636","NEWT:2853422","NEWT:1679718","NEWT:480","NEWT:67767","NEWT:46835","NEWT:109757","NEWT:582580","NEWT:1185650","NEWT:216778","NEWT:1502","NEWT:128017","NEWT:376686","NEWT:95486","NEWT:1883446","NEWT:1233435","NEWT:109760","NEWT:29031","NEWT:317047","NEWT:460523","NEWT:235443","NEWT:1757312","NEWT:85694","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:472","NEWT:940825","NEWT:529507","NEWT:1071661","NEWT:470","NEWT:5911","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:906914","NEWT:9031","NEWT:27292","NEWT:1050722","NEWT:108931","NEWT:1293497","NEWT:1055524","NEWT:150475","NEWT:267872","NEWT:172269","NEWT:9534","NEWT:5180","NEWT:256737","NEWT:9541","NEWT:8694","NEWT:33936","NEWT:8692","NEWT:2903","NEWT:185579","NEWT:13076","NEWT:1006581","NEWT:8673","NEWT:33940","NEWT:550","NEWT:554","NEWT:451516","NEWT:552","NEWT:1325291","NEWT:36185","NEWT:1054211","NEWT:1225786","NEWT:51338","NEWT:575412","NEWT:28112","NEWT:6493","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:8655","NEWT:5147","NEWT:28104","NEWT:407821","NEWT:568703","NCBITaxon:2","NEWT:568708","NEWT:986","NEWT:52641","NEWT:28532","NEWT:353152","NEWT:298176","NEWT:40674","NEWT:1194669","NEWT:443906","NEWT:519","NEWT:2510939","NEWT:7394","NEWT:6063","NEWT:1328388","NEWT:1548728","NEWT:667127","NEWT:9557","NEWT:377586","NEWT:300641","NEWT:39655","NEWT:9554","NEWT:38323","NEWT:256318","NEWT:206411","NCBITaxon:6191","NEWT:229533","NEWT:214053","NEWT:80863","NEWT:90675","NEWT:52638","NEWT:57075","NEWT:1666905","NEWT:8697","NEWT:8695","NEWT:884204","NEWT:1123869","NEWT:9544","NEWT:9545","NEWT:979","NEWT:7370","NEWT:83906","NEWT:99488","NEWT:1134506","NEWT:255470","NEWT:38783","NEWT:6426","NEWT:33090","NEWT:9935","NEWT:287889","NEWT:305959","NEWT:13489","NEWT:29829","NEWT:92867","NEWT:92866","NCBITaxon:3055","NEWT:51750","NEWT:202950","NEWT:295027","NCBITaxon:11320","NEWT:632957","NEWT:9925","NCBITaxon:9606","NEWT:90690","NEWT:1436183","NEWT:300273","NEWT:4232","NEWT:416348","NEWT:11298","NEWT:4236","NEWT:7742","NEWT:196627","NEWT:200308","NEWT:200302","NEWT:870435","NEWT:9913","NEWT:9915","NEWT:105841","NEWT:2666255","NEWT:999810","NCBITaxon:5693","NEWT:28995","NEWT:1392998","NEWT:380394","NEWT:226900","NEWT:1266738","NEWT:231490","NEWT:244704","NEWT:7725","NEWT:9909","NEWT:9902","NEWT:430615","NEWT:72664","NEWT:563041","NEWT:36599","NEWT:419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