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Beli"],"technology_type":["Mass Spectrometry","Bottom-up proteomics","Data-independent acquisition"],"software":[""],"submitter_keywords":["Sumo2/3-modified proteome","Replicative senescence","Imr90 model"],"full_dataset_link":["https://www.ebi.ac.uk/pride/archive/projects/PXD077854"],"tissue":["Fibroblast Of Lung"],"sample_protocol":["Pulldown of SUMO2/3 modified proteins was possible by conjugating mouse anti-SUMO2/3 [8A2] (1:1,000, Abcam ab81371) antibody to DynabeadsTM protein G beads (10004D, Thermo Fisher Scientific, USA). For each pulldown, a total of 5 µg of SUMO2/3 antibody was conjugated to 0.750 mg of DynabeadsTM protein G beads according to the manufacturer’s instructions. Subsequently, antibodies were crosslinked to the beads using 5 mM BS3 Crosslinker (21580, Thermo Fisher Scientific, USA), attending the manufacturer’s instructions, for a total of 30 minutes.  The cross-linking reaction was then stopped by adding 12.5 µL of 1 M TRIS-HCl buffer pH 7.5 and allowing the reaction to incubate for 15 minutes with rotation. Then, the cross-linked Dynabeads were washed 3× with PBS + 0.1% Tween-20, before being subsequently blocked with 0.5% BSA in PBS + 0.1% Tween-20, with rotation for 30 minutes at RT. After this, the beads were washed 3× with PBS + 0.1% Tween-20, before being resuspended in 100 µL of the same buffer.  The pulldown of SUMO2/3-modified proteins was carried out by incubating 100 µL of cross-linked Dynabeads, with 1 mg of protein lysate. The incubation reaction was allowed to occur overnight, at 4°C under gentle rotation. After this, the supernatant was discarded, and the beads were washed gently 3× with PBS + 0.02% Tween20. Elution of SUMO2/3-modified proteins was carried out by adding 2× NuPAGETM LDS buffer supplemented with 10 mM of DTT; and allowing the incubation to occur for 20 minutes at 70 °C with 500 r.p.m. In-gel based digestion was used, where proteins were mixed in a 1:1 volume ratio with 1×NuPAGE LDS Sample Buffer (Life Technologies) with 1 mM DTT and heated at 70°C for 10 minutes. After cooling down, proteins were alkylated using 5.5 mM chloroacetamide and allowed to incubate for 30 minutes in the dark at RT. Samples were then loaded onto Bis-Tris NuPAGE gels (Life Technologies) and allowed to run approximately 1 cm into the resolving gel, followed by fixing using a fixing solution (50% methanol, 10% acetic acid) and staining with a Colloidal Blue staining kit (Life Technologies) following the manufacturer’s instructions. Each lane was then cut into small cubes (~1 mm³) that were subsequently destained (25 mM ammonium bicarbonate, 50% ethanol) and dehydrated (100% ethanol, for 10 minutes). After dehydration, gel pieces were rehydrated with trypsin solution (T6567, Sigma, Germany; 12.5 μg/mL in 50 mM ammonium bicarbonate), and the digestion reaction was allowed to proceed overnight at 37°C. After protein trypsinization, peptide extraction buffer (30% ACN, 3% trifluoroacetic acid (TFA)) was added to stop the reaction, and peptides were extracted in three steps using successively increasing concentrations of ACN: peptide extraction buffer solution, buffer B (80% ACN, 0.5% acetic acid) and 100% ACN. Peptide-enriched supernatants were then combined and concentrated using vacuum centrifugation at 45°C to decrease ACN concentration to < 30% (v/v). Samples were then acidified using TFA solution to pH < 2 and loaded onto C18 StageTips for desalting before MS injection."],"repository":["Pride"],"quantification_method":["Not available"],"modification":[""],"data_protocol":["DIA raw files were analyzed using DIA-NN v2.02 Academia software85. We started by generating a spectral library, using the UniProtKB/Swiss-Prot file from the analysis of the DDA-data as a reference, using the DIA-NN pipeline for library generation. All libraries were generated considering the raw data needed to be analyzed. The DIA raw files were analyzed with the predicted library, allowing for N-terminal M excision, C carbamidomethylation on cysteine and 2 missed cleavages. The number of variable modifications was set to 5. Oxidation was enabled as a variable modification. Peptide range was set from 7 to 30; with precursor charges enabled from +1 to +4. A precursor range of 350 – 1,050 m/z was used, as well as a fragment ion range of 150 – 2,000 m/z. Mass tolerance was automatically inferred by DIA-NN. False discovery rates were set to 1% at both protein and peptide levels. Normalization was disabled."],"omics_type":["Proteomics"],"labhead":["Petra Beli"],"instrument_platform":[""],"labhead_affiliation":["Institute of Molecular Biology (IMB), 55128 Mainz, Germany; Institute of Developmental Biology and Neurobiology (IDN), Johannes Gutenberg University, 55128 Mainz, Germany"],"submission_type":["PARTIAL"],"species":["Homo Sapiens (human)"],"submitter_mail":["p.beli@imb-mainz.de"],"publication":["Not available"],"submitter_affiliation":["Institute of Molecular Biology (IMB), Mainz, Germany"],"submitter_country":["Germany"],"additional_accession":[]},"is_claimable":false,"name":"Alterations in proteome and proteostasis networks during replicative senescence","description":"The accumulation of senescent cells constitutes a pivotal hallmark of tissue ageing, contributing to disruption of tissue homeostasis and thus, to the development of several aging-related diseases. Despite this, our knowledge about the molecular, spatial and temporal changes that drive aging on a cellular basis remains poorly understood. To elucidate these mechanisms, we employed mass spectrometry-based proteomics to determine changes in the proteome and its post translational modifications, as well as protein solubility, during replicative senescence in human IMR90 fibroblasts. This accession contains data regarding alterations on the SUMO2/3-modified proteome during replicative senescence.","dates":{"publication":"2026-07-16","submission":"2026-04-30"},"accession":"PXD077854","cross_references":{"TAXONOMY":["NEWT:3555","NEWT:71647","NEWT:330879","NEWT:35554","NCBITaxon:12939","NEWT:32046","NEWT:544496","NEWT:59529","NEWT:2042546","NEWT:32049","NEWT:1392696","NEWT:259447","NEWT:295546","NCBITaxon:2719036","NEWT:45351","NEWT:445974","NEWT:43179","NEWT:180454","NEWT:5722","NCBITaxon:1280","NEWT:112503","NEWT:1129","NEWT:309807","NEWT:10245","NEWT:55153","NEWT:309800","NEWT:281395","NEWT:1211601","NEWT:876138","NEWT:44271","NEWT:193516","NEWT:351581","NEWT:43186","NEWT:428146","NEWT:111205","NEWT:498257","NEWT:10036","NEWT:1590","NEWT:498019","NEWT:1351","NEWT:1438992","NEWT:1352","NEWT:2649997","NEWT:1147161","NEWT:661410","NEWT:638632","NEWT:263737","NEWT:224326","NEWT:1333499","NEWT:376619","NCBITaxon:79857","NEWT:1096976","NEWT:5702","NEWT:95648","NEWT:1589","NEWT:135622","NEWT:1349","NEWT:35786","NEWT:1580","NEWT:399784","NEWT:96731","NEWT:383379","NEWT:1401257","NEWT:418106","NEWT:10029","NEWT:913645","NEWT:44491","NEWT:641809","NEWT:27933","NEWT:556484","NEWT:317447","NEWT:4688","NEWT:7955","NEWT:111225","NEWT:7719","NEWT:7959","NEWT:2261","NEWT:868565","NEWT:31156","NEWT:3112","NEWT:79329","NEWT:4442","NEWT:192875","NEWT:1214915","NEWT:12637","NEWT:59729","NEWT:2164133","NEWT:284218","NEWT:295105","NEWT:108061","NEWT:60711","NEWT:1316931","NEWT:224308","NEWT:3347","NEWT:511145","NEWT:160621","NEWT:212790","NEWT:931281","NEWT:4432","NEWT:5762","NEWT:5763","NEWT:658457","NEWT:1310161","NEWT:77133","NEWT:295358","NEWT:145481","NCBITaxon:79824","NEWT:2246","NEWT:2652724","NEWT:1912919","NEWT:5757","NEWT:68415","NCBITaxon:4563","NEWT:5755","NEWT:44688","NEWT:44689","NEWT:44447","NEWT:752555","NEWT:498211","NEWT:5759","NEWT:347256","NEWT:1736231","NEWT:5518","NEWT:398007","NEWT:1527468","NEWT:1392","NEWT:498217","NEWT:498216","NEWT:2242","NEWT:11320","NEWT:286","NEWT:391619","NEWT:246196","NEWT:287","NEWT:246197","NEWT:1367269","NEWT:145458","NEWT:633149","NEWT:1149786","NEWT:10239","NEWT:68895","NEWT:44685","NEWT:161934","NEWT:4897","NEWT:1148","NEWT:5744","NEWT:72901","NEWT:213863","NEWT:5508","NEWT:3329","NEWT:5507","NEWT:410661","NEWT:55571","NEWT:35500","NEWT:1235816","NEWT:1140","NCBITaxon:2157","NEWT:1143","NEWT:4896","NEWT:1287689","NEWT:1390","NEWT:11557","NEWT:1094343","NEWT:1462472","NEWT:1336795","NEWT:644042","NEWT:294","NEWT:1773","NEWT:1182590","NEWT:3712","NEWT:82380","NEWT:3711","NEWT:935293","NEWT:375146","NEWT:270643","NEWT:2065263","NEWT:177437","NEWT:1772","NEWT:10418","NEWT:118698","NEWT:1616117","NEWT:263","NEWT:118696","NEWT:34865","NEWT:52283","NEWT:11988","NEWT:284812","NEWT:8175","NEWT:400667","NEWT:43330","NEWT:1603293","NEWT:408169","NEWT:44664","NEWT:47946","NEWT:459758","NEWT:3702","NEWT:1245466","NEWT:243277","NEWT:1246791","NEWT:7067","NEWT:118694","NEWT:586722","NEWT:2850","NEWT:118691","NEWT:34871","NEWT:33548","NEWT:274","NEWT:7070","NCBITaxon:287","NEWT:408172","NEWT:408170","NEWT:96794","NEWT:3708","NEWT:332648","NEWT:44670","NEWT:536231","NEWT:376219","NEWT:219813","NEWT:1510","NEWT:1436733","NEWT:460519","NEWT:1247411","NEWT:1515","NEWT:572307","NEWT:1432138","NEWT:1424507","NEWT:1194599","NEWT:272844","NEWT:9483","NEWT:333760","NEWT:56636","NEWT:1513458","NEWT:1233681","NEWT:2709072","NCBITaxon:40559","NEWT:506599","NEWT:2853422","NEWT:81077","NEWT:84588","NEWT:1679718","NEWT:480","NEWT:65349","NEWT:67767","NEWT:46835","NEWT:109757","NEWT:582580","NEWT:300852","NEWT:294607","NEWT:1502","NEWT:128017","NEWT:376686","NEWT:95486","NEWT:508771","NEWT:1883446","NEWT:253","NCBITaxon:2759","NEWT:1233435","NEWT:93061","NEWT:93062","NEWT:34613","NCBITaxon:4932","NEWT:109760","NEWT:943274","NEWT:644223","NEWT:235443","NEWT:108458","NEWT:5936","NEWT:272623","NEWT:272624","NEWT:320637","NEWT:886559","NEWT:3750","NEWT:55521","NEWT:983964","NEWT:118499","NEWT:11706","NEWT:32644","NEWT:527796","NEWT:1130798","NEWT:55529","NEWT:499175","NEWT:109779","NEWT:3745","NEWT:1715989","NCBITaxon:4751","NEWT:476272","NEWT:3747","NEWT:195051","NEWT:3988","NEWT:1116234","NEWT:561007","NEWT:1255228","NEWT:649908","NEWT:410289","NEWT:373153","NEWT:3983","NEWT:203696","NEWT:352472","NEWT:1071661","NEWT:1202532","NEWT:360094","NEWT:470","NEWT:41364","NEWT:5911","NEWT:1313","NEWT:1218097","NEWT:1080348","NCBITaxon:50557","NEWT:39251","NEWT:39491","NEWT:212142","NCBITaxon:5811","NEWT:29491","NEWT:101841","NEWT:153481","NEWT:2014887","NEWT:33952","NEWT:153009","NEWT:261756","NEWT:571256","NEWT:40483","NEWT:63366","NEWT:63367","NEWT:688333","NEWT:215402","NCBITaxon:548681","NEWT:318586","NCBITaxon:5820","NEWT:1547","NEWT:272634","NEWT:198107","NEWT:9031","NEWT:27292","NEWT:1872122","NEWT:1308","NEWT:7091","NEWT:1034015","NEWT:760568","NEWT:141262","NEWT:108931","NEWT:1293497","NEWT:1055524","NEWT:4087","NEWT:9778","NEWT:306","NEWT:150475","NEWT:303","NEWT:267872","NEWT:97477","NEWT:7111","NEWT:347515","NEWT:172269","NEWT:9534","NEWT:6269","NEWT:5180","NEWT:256737","NEWT:4090","N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