{"database":"Pride","file_versions":[{"headers":{"Content-Type":["application/json"]},"body":{"files":{"Xlsx":["ftp://ftp.pride.ebi.ac.uk/pride/data/archive/2026/04/PXD059114/overview_data_files_hPOP_final.xlsx"],"Fasta":["ftp://ftp.pride.ebi.ac.uk/pride/data/archive/2026/04/PXD059114/HumanGutDietcRAP_DECOY.fasta"],"Other":["ftp://ftp.pride.ebi.ac.uk/pride/data/archive/2026/04/PXD059114/control-samples-Comet.zip","ftp://ftp.pride.ebi.ac.uk/pride/data/archive/2026/04/PXD059114/control-samples-mzML.zip","ftp://ftp.pride.ebi.ac.uk/pride/data/archive/2026/04/PXD059114/raw-data.zip","ftp://ftp.pride.ebi.ac.uk/pride/data/archive/2026/04/PXD059114/hPOP-Samples-Comet.zip","ftp://ftp.pride.ebi.ac.uk/pride/data/archive/2026/04/PXD059114/hPOP-Samples-Comet2.zip","ftp://ftp.pride.ebi.ac.uk/pride/data/archive/2026/04/PXD059114/hPOP-Samples-mzML.zip"]},"type":"primary"},"statusCode":"OK","statusCodeValue":200}],"scores":null,"additional":{"labhead_mail":["rmoritz@systemsbiology.org"],"submitter":["Ulrike Kusebauch"],"technology_type":["Data-dependent acquisition","Mass Spectrometry"],"software":["Not available"],"submitter_keywords":["Stool biospecimen","Human","Data-dependent acquisition","Dda-pasef","Timstof"],"full_dataset_link":["https://www.ebi.ac.uk/pride/archive/projects/PXD059114"],"tissue":["Feces"],"sample_protocol":["hPOP stool samples were provided in 2 mL tubes. A small portion of the stool material from each sample was transferred by spatula to a 0.2 mL vial containing four 1.4 mm ceramic beads for protein isolation. Stool samples were homogenized in 200 µL of 5% SDS, 50mM Tris-HCl (pH 8.0) using a Precellys Evolution Homogenizer (Bertin Technologies) for 6 x 30 seconds at 8500 rpm with 30 seconds resting between each cycle. Following homogenization, the samples were centrifuged twice for 15 minutes at 4100 x g, with the supernatant retained each time. Protein content was determined by bicinchoninic acid assay (BCA, Pierce). 300 µg protein from each hPOP stool sample was transferred to 96-Well S-Trap plates (ProtiFi). Proteins were denatured (90°C, 2 minutes), reduced with 5 mM DTT (30 min, 60°C), and alkylated with 10 mM iodoacetamide (30 min, room temperature). Samples were then precipitated with 350 µL 50mM Tris-HCl, 90% methanol, pH 8.0 and processed on an S-Trap plate per the manufacturer’s instructions. Samples were digested with 3 µg trypsin (Promega) in 125 µL Tris (pH 8) for 16 h at 37°C, then eluted with 80 µL 0.2% formic acid and again with 80 µL 50% acetonitrile 0.2% formic acid. The digested material was dried to completion and stored at -80°C.  All peptide samples were reconstituted in 0.1% formic acid in HPLC-grade water and analyzed using an Evosep One system (Evosep Biosytems, Denmark) coupled to a CaptiveSpray (CS) ion source (Bruker Daltonics, Germany) and a timsTOF Pro (Bruker Daltonics, Germany) mass spectrometer. The Evosep One system was operated with Buffer A, 99.9% water, 0.1% formic acid/Milli-Q water (v/v) and Buffer B, 99.9% ACN, 0.1% formic acid (v/v). 200 ng of each sample was loaded onto a disposable Evotip C18 trap (Evosep Biosytems, Denmark) as per the manufacturer’s instructions. Briefly, Evotips were rinsed with 20 µl buffer B, conditioned with 2-propanol, equilibrated with 20 µl buffer A and then loaded using centrifugal force at 700 g for 60 s. Evotips were subsequently washed with Buffer A, and then 200 μL of Buffer A was added to each tip to prevent drying. The Evosep One system was operated with a 30 Samples Per Day (30 SPD) premade method using a C18, 25 cm × 0.2 mm × 1.5 μm column (Bruker/PepSep) at room temperature. The CS source was equipped with a 20 μm emitter (Bruker Daltonics, Germany) and the following parameters were used: 1700 V capillary voltage, 3.0 L/min dry gas, and temperature set to 180°C. All samples were acquired in DDA-Parallel Accumulation Serial Fragmentation (PASEF) scan mode covering 100–1700 m/z with 10 PASEF ramps. The tims settings were 100 ms for ramp and accumulation time (100% duty cycle), resulting in 1.38 s of total cycle time. Active exclusion was enabled with a 0.2 min release. The default collision energy with a base of 0.6 1/K0 [V s/cm 2] is set at 20 eV and 1.6 1/K0 [V s/cm 2] at 63 eV was used. Isolation widths were set at 2 m/z at <700 m/z and 3 m/z at >800 m/z."],"repository":["Pride"],"quantification_method":["Not available"],"modification":[""],"data_protocol":["Raw spectral data were converted to the mzML open data format using TDF2MZML [https://github.com/mafreitas/tdf2mzml]. The spectra were analyzed using the Comet database search algorithm [PMID: 23148064] version 2021.01rev0. The FASTA sequence database consisted of proteomes obtained from UniProt (Homo sapiens, Gallus gallus, Sus scrofa, Bos taurus, Zea mays, Triticum aestivum, and Oryza sativa). These proteomes were chosen to match the sample providers and common animal and grain food proteins. Additionally, the sequence database included the Human Microbiome Project (HMP) gastrointestinal tract metaproteome. [PMID: 22699610, PMID: 28953883 PMID 32448430]  Decoy sequences were provided for all protein sequences by random shuffling of the amino acids. Database search parameters included fixed carbamidomethyl cysteine (+57.021464 Da), and variable detection of methionine oxidation (+15.9949 Da) and n-terminal protein acetylation (+42.010565 Da). Peptide-spectrum matches (PSMs) were validated using PeptideProphet [PMID: 12403597] and iProphet [PMID: 21876204] from the Trans-Proteomic Pipeline version 6.0.0 [PMID: 36648445]. Protein inference was performed using ProteinProphet [PMID: 14632076]. Protein quantification was performed using StPeter version 1.4.0 [PMID: 29400476], with a minimum protein and PSM probability of 0.9, allowance for degenerate peptides, and scaled using a sample load value of 1. Protein quantities included in the final data matrix were filtered using a global 1% FDR at the protein level."],"omics_type":["Proteomics"],"labhead":["Robert L."],"instrument_platform":[""],"labhead_affiliation":["Institute for Systems Biology, Seattle, WA, USA"],"submission_type":["PARTIAL"],"species":["Homo Sapiens (human)"],"publication":["Not available"],"submitter_mail":["ukusebauch@systemsbiology.org"],"submitter_affiliation":["Institute for Systems Biology, Seattle, WA, USA"],"submitter_country":["United States"],"additional_accession":[]},"is_claimable":false,"name":"Proteomic analysis of hPOP stool samples","description":"How humans differ from one another at the molecular level and the impact of ethnicity, age and environmental factors such as diet and geography are poorly understood. To address this gap, we analyzed 322 nominally healthy individuals from diverse ethnic backgrounds located in distinct geographical regions, using (1) deep multi-omic profiling - including genome sequencing, transcriptomics, glycomics, targeted and untargeted metabolomics, lipidomics, proteomics, 16S microbiota sequencing, metaproteomics, metallomics, and metabolic hormone and pathogen profiling, (2) clinical data collection, and (3) comprehensive lifestyle surveys. Our integrative analysis revealed several key findings: associations between molecular profiles (transcripts, proteins, metabolites, lipids, glycans, and microbiota) and age, ethnicity, and geographical residence (including diet) among healthy individuals; ethnicity-specific genetic polymorphisms in key genes governing whole-body homeostasis, particularly in lipid/carbohydrate metabolism and neuroinflammation pathways; and variation in the abundance of key proteins such as the TERF2 interacting protein (TERF2IP) which is critical for cellular aging, genome stability, and protection against chromosomal degradation. These findings hold significant translational value, particularly in understanding ethnicity as a risk factor for autoimmune diseases. Many ethnicity-associated molecules identified in our study interact with existing therapeutic drugs, highlighting the potential need for ethnicity-specific drug dosing adjustments. Furthermore, our results suggest that geographical factors influence ethnic patterns, which may, in turn, affect longevity. individuals with European ancestry exhibited a lower phenotypic age, while those of Asian ancestry showed a higher phenotypic age when living outside their ancestral regions. By providing both a comprehensive baseline dataset of multi-omic variation in healthy individuals and critical insights into the molecular basis of human diversity, our study enhances our understanding of how environmental and genetic factors influence phenotypic and molecular traits and potentially augments individualized health approaches.","dates":{"publication":"2026-04-10","submission":"2024-12-21"},"accession":"PXD059114","cross_references":{"TAXONOMY":["NEWT:330879","NEWT:108018","NEWT:377960","NEWT:1131","NEWT:35795","NEWT:2042546","NEWT:259447","NEWT:295546","NEWT:1334565","NEWT:625351","NEWT:258594","NCBITaxon:1280","NEWT:1126","NEWT:112503","NEWT:1129","NEWT:309807","NEWT:59753","NEWT:89184","NEWT:309800","NEWT:281395","NEWT:247480","NEWT:1211601","NEWT:1323429","NEWT:876138","NEWT:44271","NEWT:193516","NEWT:2315688","NEWT:1367493","NEWT:592321","NEWT:1073882","NEWT:6819","NEWT:46429","NEWT:475174","NEWT:1599","NEWT:428386","NEWT:1117","NEWT:498257","NEWT:10036","NEWT:32264","NEWT:12215","NEWT:1590","NEWT:10034","NEWT:33114","NEWT:1596","NEWT:2736291","NEWT:661410","NEWT:638632","NEWT:224326","NEWT:294671","NEWT:376619","NEWT:1748418","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:656061","NEWT:37965","NEWT:656064","NEWT:1582","NEWT:3760","NEWT:1584","NEWT:2086595","NEWT:36630","NEWT:1585","NEWT:383379","NEWT:381198","NEWT:418106","NCBITaxon:12721","NEWT:10029","NEWT:913645","NEWT:641809","NEWT:44250","NEWT:1392488","NEWT:38815","NEWT:317447","NEWT:4688","NEWT:3357","NEWT:7719","NEWT:29073","NEWT:5530","NEWT:868565","NEWT:135674","NEWT:1562698","NEWT:79329","NEWT:30069","NEWT:12637","NEWT:59729","NEWT:2164133","NEWT:295105","NEWT:2767846","NEWT:314344","NEWT:108061","NEWT:60711","NEWT:2020713","NEWT:224308","NEWT:3347","NEWT:49928","NEWT:1220585","NEWT:3349","NEWT:160621","NEWT:2502409","NEWT:212790","NEWT:110368","NEWT:55350","NEWT:1916073","NEWT:33103","NEWT:1310161","NEWT:1230383","NEWT:77133","NEWT:145481","NEWT:1310165","NEWT:29058","NEWT:114727","NCBITaxon:79824","NEWT:468822","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:342808","NEWT:2579791","NEWT:614666","NEWT:44685","NEWT:161934","NEWT:1148","NEWT:5508","NEWT:3329","NEWT:419481","NEWT:5507","NEWT:260499","NEWT:410661","NEWT:329854","NEWT:1140","NCBITaxon:2157","NEWT:2066563","NEWT:2649329","NEWT:1143","NEWT:99802","NEWT:1211620","NEWT:1287689","NEWT:1094343","NEWT:1462472","NEWT:1336795","NEWT:644042","NEWT:150808","NEWT:709964","NEWT:242605","NEWT:1182590","NEWT:3712","NEWT:3711","NEWT:156974","NEWT:3714","NCBITaxon:37162","NEWT:3715","NEWT:2065263","NEWT:199225","NEWT:1549675","NEWT:177437","NEWT:717966","NEWT:9014","NEWT:10891","NEWT:10418","NEWT:118698","NEWT:1616117","NEWT:118696","NEWT:34865","NEWT:52283","NEWT:284812","NEWT:8175","NEWT:43330","NEWT:1603293","NEWT:29022","NEWT:44664","NEWT:43346","NEWT:3701","NEWT:3702","NEWT:1245466","NEWT:3704","NEWT:244366","NEWT:96795","NEWT:1246791","NEWT:176597","NEWT:118694","NEWT:2850","NEWT:118691","NEWT:9005","NEWT:34871","NEWT:9006","NEWT:33548","NEWT:52299","NEWT:1883426","NEWT:8165","NEWT:96794","NEWT:10401","NEWT:10886","NEWT:3708","NEWT:332648","NEWT:44670","NEWT:926571","NEWT:536231","NEWT:376219","NEWT:219813","NEWT:1510","NEWT:1955249","NEWT:215451","NEWT:460519","NEWT:1515","NEWT:572307","NEWT:1432138","NEWT:2714896","NEWT:1424507","NEWT:1194599","NEWT:272844","NEWT:1348799","NEWT:1608994","NEWT:33995","NEWT:55784","NEWT:1303443","NEWT:485","NEWT:329032","NEWT:56636","NEWT:2499525","NEWT:55307","NEWT:1122981","NEWT:2853422","NEWT:1122984","NEWT:487","NEWT:1538553","NEWT:1679718","NEWT:480","NEWT:67767","NEWT:45972","NEWT:2830","NEWT:46835","NEWT:109757","NEWT:582580","NEWT:1185650","NEWT:216778","NEWT:1502","NEWT:128017","NEWT:2836","NEWT:376686","NEWT:237610","NEWT:95486","EFO:0001352","NEWT:219809","NEWT:219808","NEWT:119562","NEWT:1883446","NEWT:31345","NEWT:1233435","NEWT:118223","NEWT:11757","NEWT:141469","NEWT:987059","NEWT:109760","NEWT:29031","NEWT:317047","NEWT:1509","NEWT:491","NEWT:460523","NEWT:45954","NEWT:1578","NEWT:235443","NEWT:1579","NEWT:1757312","NEWT:85694","NEWT:109786","NEWT:1922344","NEWT:108458","NEWT:5936","NEWT:85692","NEWT:321967","NEWT:320637","NEWT:1078283","NEWT:3750","NEWT:983964","NEWT:11706","NEWT:32644","NEWT:527796","NEWT:1392498","NEWT:279010","NEWT:291146","NEWT:499175","NEWT:109779","NEWT:3745","NEWT:1715989","NCBITaxon:4751","NEWT:3747","NEWT:278161","NEWT:1116234","NEWT:415978","NEWT:235455","NEWT:1255228","NEWT:410289","NEWT:191701","NEWT:373153","NEWT:1640278","NEWT:9054","NEWT:1092586","NEWT:472","NEWT:940825","NEWT:29459","NEWT:529507","NEWT:1071661","NEWT:28128","NEWT:470","NEWT:41122","NEWT:5911","NEWT:1328300","NEWT:748693","NCBITaxon:50557","NEWT:39251","NEWT:29491","NEWT:911022","NEWT:101841","NEWT:446","NEWT:153481","NEWT:2014887","NEWT:717989","NEWT:33952","NEWT:1255232","NEWT:11723","NEWT:445","NEWT:153009","NEWT:1004879","NEWT:261756","NEWT:166314","NEWT:166313","NEWT:63366","NEWT:63367","NEWT:29488","NEWT:1871049",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