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Parker"],"technology_type":["Mass Spectrometry","Bottom-up proteomics"],"software":[""],"submitter_keywords":["Pdia6","Iodo-tmt"],"full_dataset_link":["https://www.ebi.ac.uk/pride/archive/projects/PXD075204"],"tissue":["Skeletal Muscle"],"sample_protocol":["Global quantification of cysteine oxidation was performed using an iodoTMT isobaric labelling kit (Thermo Scientific; #90101) according to their product protocol. Briefly, gastrocnemius muscles over-expressing GFP or FLAG-PDIA6 (n=3 each) were powdered under liquid nitrogen and lysed in 2% SDS in 100mM HEPES pH 6.9 containing 150mM NaCl, 100mM NEM and 5mM EDTA via tip probe sonication. NEM labelling was performed for 1 hour in the dark after which methanol-chloroform precipitation was performed to remove NEM. Protein was then normalised and reduced with 2% SDS in 100mM HEPES pH 6.9 containing 150mM NaCl and 50mM TCEP for 30 mins at 45 °C. Methanol-chloroform precipitation was performed to remove TCEP where protein was normalised again. IodoTMT reagents were added to each sample where labelling occurred for 1 hour at 37 °C in the dark. The reaction was quenched with 20mM DTT for 15 minutes at 37 °C in the dark after which samples were combined. A final chloroform methanol-precipitation was performed, and protein pellet was resuspended in 1.2M Urea in 100mM HEPES pH7.5. Proteins were digested with 1:50 ratio of trypsin and LysC overnight at 37 °C in the dark shaking at 2000RPM. Peptides were acidified to 0.1% TFA and loaded onto a HLB-SPE column for cleanup then lyophilized via a vacuum concentrator. The purified peptides were resuspended in 2% acetonitrile in 0.1% TFA and separated into 12 fractions by neutral phase C18BEH HPLC. peptides were analyzed on a Vanquish Neo nanoHPLC, coupled to an Orbitrap Astral mass spectrometer (ThermoFischer Scientific) via electrospray ionization in positive mode with 1.9 kV at 290 °C and RF set to 40%. Separation was achieved on a 5 cm × 75 µm uPAC column (Thermo Scientific) over 17min at a flow rate of 750 nL/min. The peptides were eluted over a linear gradient of 3–40% Buffer B (Buffer A: 0.1% v/v formic acid; Buffer B: 80% v/v acetonitrile, 0.1% v/v FA) and the column was maintained at 50 °C. The instrument was operated in data-dependent acquisition mode with an MS1 spectrum acquired over the mass range 350–1500 m/z (120,000 resolution, 300% automatic gain control (AGC) and 50 ms maximum injection time) followed by MS/MS via HCD fragmentation with a cycle time of 1 seconds where only peptides with charge state 2-7 were selected with 0.5 m/z isolation window (collision energy of 35%, 100% AGC, maximum injection time set to 20 ms)."],"repository":["Pride"],"quantification_method":["Not available"],"modification":[""],"data_protocol":["IodoTMT data was searched against a custom mouse skeletal muscle database with SequestHT within Proteome Discoverer v2.5.0.4 (Eng et al., 1994). The precursor MS tolerance was set to 20 ppm and the MS/MS tolerance was set to 0.02 Da with a maximum of 2 miss-cleavage. The peptides were searched with oxidation of methionine, iodoTMT and Nethylmaleimide of Cysteine set as variable modification. Peptide spectral matches were filtered to 1% FDR using a target/decoy approach with Percolator (Kall et al., 2007). Quantification was performed with the reporter ion quantification node for TMT quantification in Proteome Discoverer. TMT precision was set to 20 ppm and corrected for isotopic impurities. Only spectra with < 50% co-isolation interference were used for quantification with an average signal-to-noise filter of > 10."],"omics_type":["Proteomics"],"labhead":["Benjamin Parker"],"instrument_platform":[""],"labhead_affiliation":["The University of Melbourne"],"submission_type":["PARTIAL"],"species":["Mus Musculus (mouse)"],"submitter_mail":["ben.parker@unimelb.edu.au"],"publication":["Not available"],"submitter_affiliation":["The University of Melbourne"],"submitter_country":["Australia"],"additional_accession":[]},"is_claimable":false,"name":"Cysteine Redox proteomics of PDIA6 over-expressing skeletal muscle","description":"To investigate the role of PDIA6 on changes to specific cysteine residues with altered redox status and insulin senstivity, we overexpressed either GFP control or FLAG-PDIA6 throughout the musculature of 4-day old male C57BL/6J mice via intraperitoneal injection with MyoAAV (Fig. 5F). At 8 weeks of age, mice were placed on a HFD for 12 weeks to induce insulin resistance. Muscle lysates were were labelled with iodoTMT and analysed by LC-MS/MS","dates":{"publication":"2026-05-29","submission":"2026-03-04"},"accession":"PXD075204","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: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: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: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: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","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: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","NEWT:4092","NEWT:9541","NEWT:8694","NEWT:4093","NEWT:33936","NEWT:4096","NEWT:8692","NEWT:2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