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Huesgen"],"technology_type":["Mass Spectrometry","Data-independent acquisition","Bottom-up proteomics"],"software":[""],"submitter_keywords":["A single-amino-acid cleavage controls global mitochondrial complex integrity"],"full_dataset_link":["https://www.ebi.ac.uk/pride/archive/projects/PXD070326"],"tissue":["Cell Culture"],"sample_protocol":["Proteins were extracted from isolated wt and ICP55⁻/⁻ mitochondria in denaturing extraction buffer (4% SDS, 100 mM HEPES, 5 mM EDTA, pH 7.5). N-terminal peptides were enriched essentially as described (PMID: 39455217). In brief, proteins were reduced, alkylated and labelled by reductive dimethylation with heavy formaldehyde (¹²CD₂O for digestion with trypsin and ¹3CD₂O for digestion with legumain). Each sample was independently digested with trypsin (1:100 w/w) and legumain (1:50 w/w) overnight to increase N-terminome coverage. 10 μg of digested peptides were retained to determine label efficiency (pre-HUNTER). Free -amines were subsequently modified with undecanal and separated from N-terminal peptides by retention on reversed-phase spin cartridges (HR-X spin cartridges, Macherey and Nagel). N-terminal peptides remained in the flow-through, were concentrated in a vacuum concentrator, desalted using homemade C18 stage tips with three layers and reconstituted in 0.1% formic acid. For trypsin-digested samples, 500 ng enriched peptides were loaded on Evotips (Evosep) and separated on an Evosep One system coupled online to an Orbitrap Exploris 480 mass spectrometer (Thermo) using a 44-min gradient (30 SPD workflow, Evosep, solvent A 0.1% formic acid and solvent B 100% acetonitrile, 0.1% formic acid) with a nano-LC column (EV1137, 15 cm × 150 µm performance column, Evosep) maintained at 40°C with a column oven (PRSVO-V2, Sonation). Peptides were ionized with a stainless-steel emitter with an integrated liquid junction (EV1072, Evosep) and mounted into an EasySpray adapter (EV1072, Evosep) in a NanoSpray Flex ion source (Thermo). A spray voltage of +2.0 kV was applied, and the ion transfer tube was set to 275 °C. For legumain-digested samples, 500 ng peptides were analyzed with an UltiMate 3000 RSLCnano system (Thermo) coupled online to an Exploris 480 mass spectrometer (Thermo). Peptides were loaded onto a PAC™ C18 trapping column (Thermo) with a flow rate of 10 l/min, then separated on a μPAC™ C18 pillar array column (50 cm bed length, Thermo) using a binary solvent system (solvent A , 0.1% formic acid; solvent B 86% acetonitrile, 0.1% formic acid. The flow rate was 500 nl/min and peptides were eluted with a gradient of 1-4% B in 2 min, 4-25% B in 24 min, 25-44% B in 12 min, 44-90% B in 2 min, followed by 4 min at 90% B. Peptides were ionized with a Nanospray Flex ion source equipped with a PAC Flex iON Connector (Thermo) and a fused silica emitter (20 m inner diameter, 10 m inner diameter of the tip, 360 m outer diameter; MicroOmics Technologies LLC), applying a +1.8 kV source voltage and an ion transfer tube temperature of 275°C. Data were acquired in data-independent acquisition (DIA) mode. Each cycle comprised one MS1 survey scan (m/z 350–1400, 120,000 resolution, RF lens 40%, AGC target 300%, maximum injection time 45 ms) followed by MS2 scans (profile mode, 30,000 resolution, RF lens 50%, AGC target 1000%, maximum injection time 54 ms, variable window size with 1 m/z overlap)."],"repository":["Pride"],"quantification_method":["Not available"],"modification":[""],"data_protocol":["Raw files were converted to mzML format using MSConvert (v3.0.21229-9668f52) with the options ‘Write index’, ‘TPP compatibility’, and ‘Use zlib compression’ enabled. The default ‘peakPicking’ filter was applied first, followed by the ‘zeroSamples’ filter. Converted spectra files were processed using the DIA-Umpire signal extraction module as implemented in FragPipe (v22.0) followed by MSFragger searches considering semi-specific cleavage (variable N-terminus, C-terminal Arg for trypsin and C-terminal Asn or Asp for legumain digested samples) with up to two missed cleavages, peptide length between 7 and 45 amino acids in a mass range of 200–5000 Da, with up to five variable modifications per peptide. Dimethylation at protein N-termini and Lys side chains (+32.0564 Da for trypsin, +34.0631 Da for legumain) and Cys carbamidomethylation (+57.0215 Da) were set as a fixed modifications, and Met oxidation (+15.9949 Da), acetylation of peptide N termini and lysines (+9.9542 Da for trypsin, +7.9474 Da for legumain), N-terminal loss of ammonia, N-terminal pyroglutamate formation from glutamine (-49.083 Da for trypsin, -51.090 Da for legumain) or glutamic acid (-50.067 Da for trypsin, -52.074 Da for legumain), and proline monomethylation (-16.028 Da for trypsin, -17.032 Da for legumain) as variable modifications. Peptide and protein identifications were filtered at 1% FDR at both the peptide-spectrum match and protein level using Philosopher. Quantification was performed with DIA-NN as implemented in FragPipe with unrelated runs enabled, N-terminal peptide intensities were used for downstream statistical analysis."],"omics_type":["Proteomics"],"labhead":["Prof. Dr. Pitter Huesgen"],"instrument_platform":[""],"labhead_affiliation":["University of Freiburg Faculty of Biology Biochemistry and Functional Proteomics  Schänzlestrasse 1 79104 Freiburg Germany"],"submission_type":["PARTIAL"],"species":["Homo Sapiens (human)"],"publication":["Not available"],"submitter_mail":["pitter.huesgen@biologie.uni-freiburg.de"],"submitter_affiliation":["University of Freiburg\nFaculty of Biology\nBiochemistry and Functional Proteomics"],"submitter_country":["Germany"],"additional_accession":[]},"is_claimable":false,"name":"N-terminome data for \"A single-amino-acid cleavage controls global mitochondrial complex integrity\"","description":"Mitochondrial proteostasis depends on precise N-terminal processing of imported precursor proteins, and defects in this maturation step are implicated in severe human diseases, yet the functional impact in humans remain unclear. This HUNTER N-terminome dataset identify over one hundred human ICP55 substrates in mitochondria isolated from wt and ICP55-deficient HEK293T cells","dates":{"publication":"2026-07-22","submission":"2025-11-05"},"accession":"PXD070326","cross_references":{"TAXONOMY":["NEWT:330879","NEWT:377960","NEWT:1131","NEWT:35795","NEWT:2042546","NEWT:259447","NEWT:295546","NCBITaxon:2719036","NEWT:1334565","NEWT:625351","NEWT:258594","NCBITaxon:1280","NEWT:112503","NEWT:1129","NEWT:309807","NEWT:59753","NEWT:89184","NEWT:309800","NEWT:281395","NEWT:247480","NEWT:1211601","NEWT:876138","NEWT:44271","NEWT:193516","NEWT:2315688","NEWT:592321","NEWT:1073882","NEWT:6819","NEWT:112509","NEWT:475174","NEWT:111205","NEWT:1599","NEWT:428386","NEWT:1117","NEWT:498257","NEWT:10036","NEWT:32264","NEWT:12215","NEWT:1590","NEWT:10034","NEWT:33114","NEWT:1596","NEWT:661410","NEWT:638632","NEWT:224326","NEWT:376619","NCBITaxon:79857","NEWT:1096976","NEWT:3765","NEWT:3767","NEWT:1589","NEWT:135622","NEWT:4615","NEWT:672127","NEWT:35786","NEWT:1580","NEWT:399784","NEWT:96731","NEWT:656064","NEWT:1582","NEWT:3760","NEWT:1584","NEWT:36630","NEWT:1585","NEWT:383379","NEWT:381198","NEWT:418106","NCBITaxon:12721","NEWT:10029","NEWT:913645","NEWT:2492962","NEWT:641809","NEWT:1392488","NEWT:38815","NEWT:317447","NEWT:4688","NEWT:111225","NEWT:7719","NEWT:5530","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:49928","NEWT:3349","NEWT:160621","NEWT:212790","NEWT:110368","NEWT:55350","NEWT:1916073","NEWT:1310161","NEWT:77133","NEWT:145481","NEWT:1310165","NEWT:29058","NCBITaxon:79824","NEWT:1912919","NEWT:410658","NEWT:5516","NEWT:44688","NEWT:44689","NEWT:370038","NEWT:498211","NEWT:345076","NEWT:347256","NEWT:5518","NEWT:398007","NEWT:1527468","NEWT:498217","NEWT:498216","NEWT:11320","NEWT:246196","NEWT:246197","NEWT:145458","NEWT:1310173","NEWT:200795","NEWT:37919","NEWT:2579791","NEWT:44685","NEWT:161934","NEWT:156587","NEWT:1148","NEWT:5508","NEWT:3329","NEWT:419481","NEWT:5507","NEWT:260499","NEWT:410661","NEWT:1140","NCBITaxon:2157","NEWT:2066563","NEWT:2649329","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:270643","NEWT:3715","NEWT:1538","NEWT:2065263","NEWT:177437","NEWT:717966","NEWT:10891","NEWT:10418","NEWT:118698","NEWT:1616117","NEWT:118696","NEWT:34865","NEWT:52283","NEWT:28174","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:9006","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:9483","NEWT:485","NEWT:56636","NEWT:2709072","NEWT:2499525","NEWT:2853422","NEWT:487","NEWT:1538553","NEWT:1679718","NEWT:480","NEWT:67767","NEWT:45972","NEWT:46835","NEWT:109757","NEWT:582580","NEWT:1185650","NEWT:216778","NEWT:294607","NEWT:1502","NEWT:128017","NEWT:2836","NEWT:376686","NEWT:237610","NEWT:95486","EFO:0001352","NEWT:119562","NEWT:1883446","NEWT:1233435","NEWT:118223","NEWT:11757","NEWT:987059","NEWT:109760","NEWT:29031","NEWT:317047","NEWT:1509","NEWT:491","NEWT:460523","NEWT:45954","NEWT:235443","NEWT:1579","NEWT:1757312","NEWT:85694","NEWT:109786","NEWT:108458","NEWT:5936","NEWT:85692","NEWT:321967","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:415978","NEWT:1255228","NEWT:410289","NEWT:191701","NEWT:373153","NEWT:1640278","NEWT:472","NEWT:940825","NEWT:29459","NEWT:529507","NEWT:1071661","NEWT:28128","NEWT:470","NEWT:41122","NEWT:5911","NEWT:748693","NCBITaxon:50557","NEWT:39251","NEWT:29491","NEWT:101841","NEWT:446","NEWT:153481","NEWT:2014887","NEWT:33952","NEWT:11723","NEWT:445","NEWT:153009","NEWT:261756","NEWT:166313","NEWT:63366","NEWT:63367","NEWT:29488","NEWT:215402","NEWT:1547","NEWT:1290454","NEWT:3726","NEWT:906914","NEWT:81231006","NEWT:28141","NEWT:9031","NEWT:27292","NEWT:1050722","NEWT:3728","NEWT:108931","NEWT:1293497","NEWT:1055524","NEWT:548","NEWT:150475","NEWT:267872","NEWT:381666","NEWT:172269","NEWT:9534","NEWT:2126348","NEWT:5180","NEWT:256737","NEWT:9541","NEWT:8694","NEWT:33936","NEWT:8692","NEWT:542","NEWT:2903","NEWT:185579","NEWT:872227","NEWT:13076","NEWT:2587410","NEWT:207340","NEWT:1006581","NEWT:8673","NEWT:33940","NEWT:2126337","NEWT:550","NEWT:10804","NEWT:33941","NEWT:554","NEWT:451516","NEWT:552","NEWT:1325291","NEWT:42410","NEWT:36185","NEWT:1054211","NEWT:1225786","NEWT:771870","NEWT:51338","NEWT:9995","NEWT:575412","NEWT:28112","NEWT:49012","NEWT:28110","NEWT:6493","NEWT:10832","NEWT:6494","NEWT:6491","NEWT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