{"database":"Pride","file_versions":[{"headers":{"Content-Type":["application/json"]},"body":{"files":{"Other":["ftp://ftp.pride.ebi.ac.uk/pride/data/archive/2026/06/PXD047294/Albana_Mammals_Search.zip","ftp://ftp.pride.ebi.ac.uk/pride/data/archive/2026/06/PXD047294/Gymet_Mammals_Search.zip","ftp://ftp.pride.ebi.ac.uk/pride/data/archive/2026/06/PXD047294/HPM_20180212_Mammal_RawData.zip","ftp://ftp.pride.ebi.ac.uk/pride/data/archive/2026/06/PXD047294/CONVERSE_Mammals_Search.zip","ftp://ftp.pride.ebi.ac.uk/pride/data/archive/2026/06/PXD047294/Marshal_Mammals_Search.zip","ftp://ftp.pride.ebi.ac.uk/pride/data/archive/2026/06/PXD047294/Tallouse_Mammal_RawData.zip","ftp://ftp.pride.ebi.ac.uk/pride/data/archive/2026/06/PXD047294/Marshal_Mammal_RawData.zip","ftp://ftp.pride.ebi.ac.uk/pride/data/archive/2026/06/PXD047294/SMURF_Mammals_Search.zip","ftp://ftp.pride.ebi.ac.uk/pride/data/archive/2026/06/PXD047294/HPM_20180212_Mammals_Search.zip","ftp://ftp.pride.ebi.ac.uk/pride/data/archive/2026/06/PXD047294/Solstice_Mammals_Search.zip","ftp://ftp.pride.ebi.ac.uk/pride/data/archive/2026/06/PXD047294/Avatar_Mammal_RawData.zip","ftp://ftp.pride.ebi.ac.uk/pride/data/archive/2026/06/PXD047294/Gymet_Mammal_RawData.zip","ftp://ftp.pride.ebi.ac.uk/pride/data/archive/2026/06/PXD047294/Albana_Mammal_RawData.zip","ftp://ftp.pride.ebi.ac.uk/pride/data/archive/2026/06/PXD047294/Applicable_Mammal_RawData.zip","ftp://ftp.pride.ebi.ac.uk/pride/data/archive/2026/06/PXD047294/HPM_20180116_Mammal_Search.zip","ftp://ftp.pride.ebi.ac.uk/pride/data/archive/2026/06/PXD047294/Best_Mammal_RawData.zip","ftp://ftp.pride.ebi.ac.uk/pride/data/archive/2026/06/PXD047294/SIXPACK_Mammals_Search.zip","ftp://ftp.pride.ebi.ac.uk/pride/data/archive/2026/06/PXD047294/Octane_Mammals_Search.zip","ftp://ftp.pride.ebi.ac.uk/pride/data/archive/2026/06/PXD047294/HPM_20180116_Mammal_RawData.zip","ftp://ftp.pride.ebi.ac.uk/pride/data/archive/2026/06/PXD047294/CONVERSE_Mammal_RawData.zip","ftp://ftp.pride.ebi.ac.uk/pride/data/archive/2026/06/PXD047294/Tallouse_Mammals_Search.zip","ftp://ftp.pride.ebi.ac.uk/pride/data/archive/2026/06/PXD047294/Apprentic_Mammal_RawData.zip","ftp://ftp.pride.ebi.ac.uk/pride/data/archive/2026/06/PXD047294/Best_Mammals_Search.zip","ftp://ftp.pride.ebi.ac.uk/pride/data/archive/2026/06/PXD047294/Applicable_Mammals_Search.zip","ftp://ftp.pride.ebi.ac.uk/pride/data/archive/2026/06/PXD047294/Utah_Mammal_RawData.zip","ftp://ftp.pride.ebi.ac.uk/pride/data/archive/2026/06/PXD047294/SOLSTICE_Mammal_RawData.zip","ftp://ftp.pride.ebi.ac.uk/pride/data/archive/2026/06/PXD047294/SIXPACK_Mammal_RawData.zip","ftp://ftp.pride.ebi.ac.uk/pride/data/archive/2026/06/PXD047294/Octane_Mammal_RawData.zip","ftp://ftp.pride.ebi.ac.uk/pride/data/archive/2026/06/PXD047294/Utah_Mammals_Search.zip","ftp://ftp.pride.ebi.ac.uk/pride/data/archive/2026/06/PXD047294/Avatar_Mammals_Search.zip","ftp://ftp.pride.ebi.ac.uk/pride/data/archive/2026/06/PXD047294/Apprentic_Mammals_Search.zip","ftp://ftp.pride.ebi.ac.uk/pride/data/archive/2026/06/PXD047294/SMURF_Mammal_RawData.zip"]},"type":"primary"},"statusCode":"OK","statusCodeValue":200}],"scores":null,"additional":{"labhead_mail":["george.katselis@usask.ca"],"submitter":["Paulos Chumala"],"technology_type":["Data-dependent acquisition","Mass Spectrometry"],"software":["Not available"],"submitter_keywords":["Bull fertility","Proteomics analysis","Sperm head plasma membrane","Fertility biomarkers"],"full_dataset_link":["https://www.ebi.ac.uk/pride/archive/projects/PXD047294"],"tissue":["Sperm","Endosperm"],"sample_protocol":["Fresh double ejaculates were obtained from 16 Holstein Friesian bulls with known field fertility, classified as average-to-high fertility (ATH, n=8) and average-to-low fertility (ATL, n=8) based on the international multi-factor BFI. The HPM was obtained according to an established procedure [18-20], performing all steps at room temperature unless specified otherwise. The final pellet was subjected to nitrogen cavitation in a cell disruption Parr cavitation unit (Parr instrument company, IL, USA; 650 psi, 10 min) introducing nitrogen gas over 90 sec to a final pressure of 650 psi, holding for 10 min and then releasing pressure over 90 sec. Finally, the cavitate was centrifuged at 800 x g for 10 min. Phase partition tubes were prepared by mixing 3.94 g 20% dextran (1:5 g: g; dextran: water), 1.97 g 40 % PEG (1:2.5 g: g; PEG: water), 0.19 g PBS (in 1000 mL water), 2.42 g 1.5 M sucrose (in 100 mL water), covering and keeping in fridge overnight. The samples were homogenized, aliquoted into eppendorf tubes, covered with nitrogen gas, snap frozen in liquid nitrogen, and stored at -80 ºC.   HPM protein concentration, was determined by bicinchoninic acid analysis using the BioTek ELx808 (BioTek- Instruments Inc., VT, USA) multi detection plate reader (wavelength 562 nm) and BSA as the standard protein [21]. Triplicates from each HPM digested into peptides using in-solution trypsin digestion protocol [21, 22]. Peptide fractionation, using SCX Spin Tips was incorporated to enhance sample resolution prior to MS/MS analysis [24, 25]. Dried SCX 9 fractions containing tryptic peptides were, vortexed, centrifuged and  transferred to LC-MS/MS analysis (Brandt et al., 2019; Nair et al., 2021; Koziy et al., 2022). An Agilent 6550 iFunnel Q-TOF mass spectrometer (Agilent Technologies, Mississauga, ON, CA) coupled with liquid chromatography [28] were used (Agilent Technologies, Mississauga, ON, CA) [23].   Mass spectra were acquired in positive ion mode with defined parameters, including mass range and scan rate. Auto MS/MS was performed with a set threshold for precursor ion selection and exclusion [23]"],"repository":["Pride"],"quantification_method":[""],"modification":[""],"data_protocol":["Database Search and Analysis      MS/MS spectra were obtained and searched against a Mammals species protein database (Swiss Prot, UniProt release 2020_06) using the Agilent Spectrum Mill search engine. The search parameters included trypsin as the enzyme for protein digestion, specific cleavage rules, and fixed modifications of cysteine by carbamidomethylation. Variable modifications were applied in four stages, incorporating various post-translational modifications. Validated hits from each stage were used for subsequent searches. The Spectrum Mill validation was performed at both peptide and protein levels with 1% FDR and peptide sequence length [22]. Furthermore, each identified DAP underwent manual validation using Mass Hunter software with a mass difference cutoff ∆m < 10 PPM to ensure accuracy. A comparison was made between the identified proteins in ATH (n=8) and ATL (n=8) bulls. Proteins present in both groups were categorized as common proteins, while proteins exclusive to one group were considered unique to that fertility group [23]. Statistical Analysis and Protein Network Analysis    For this study, three bulls with the highest BFI were labeled as high fertility (HF; BFI of 105,105,107), and three with the lowest BFI were designated as low fertility (LF; BFI of 88,92,94). Statistical analysis involved t-tests and was conducted using MPP software. To ensure statistical significance, the Benjamini and Hochberg correction was applied with an FDR threshold of < 1%, and a p-value cutoff of < 0.05. Proteins were classified as up- or down-regulated based on a fold change (FC) of ≥ 2 in spectral intensities between the compared groups. The resulting 67 significantly expressed proteins (DAPs) were manually validated using Mass Hunter software.    To enhance data interpretation, accession numbers of significant DAPs from multiple species were converted into homologous proteins of Homo sapiens for further protein-protein interaction (PPI) and networking analysis. Subnetworks of up- and down-regulated DAPs with all identified non-differentially abundant proteins (non-DAPs) in high-fertility HPM were constructed using Cytoscape. PPI networks among DAPs were generated through STRING analysis, which provides known and predicted PPI. A Cytoscape analysis was performed to construct a complete interactome of HPM. The most significant clusters within the DAPs' STRING PPI networks were identified using the molecular complex detection (MCODE) plugin in Cytoscape.    Linear regression analysis was conducted with SAS statistical software (SAS; version 9.3; SAS Institute, Inc Cary, NC) on the intensities of the DAPs relative to BFI. The analysis was first performed for the HF and LF groups (n=3 per group) and then for the entire population of bulls (n=16). The deterministic model used in equation (4.1) was; y=a+bx…………………………………………………………………………… (4.1) where a= intercept, b is = slope of the regression line."],"omics_type":["Proteomics"],"labhead":["Dr. George Katselis"],"instrument_platform":[""],"submission_type":["PARTIAL"],"labhead_affiliation":["Assistant Professor, Mass spectrometry Facility, Canadian Centre for Health and Safety in Agriculture, College of Medine, University of Saskatchewan."],"species":["Bos Taurus (bovine)"],"submitter_mail":["pab776@mail.usask.ca"],"publication":["41932951 Imran M, Buhr MM, Chumala P, Katselis GS. Comprehensive proteomics analysis of bovine sperm head plasma membrane associated with fertility. Sci Rep. 2026 16(1):15930 10.1038/s41598-025-34626-8"],"curator_keywords":["Human proteome project","Biology/disease-driven human proteome project (b/d-hpp)"],"submitter_affiliation":["Canadian Centre for Rural and Agricultural Heatlh , College of Medicine, University of Saskatchewan"],"submitter_country":["Canada"],"pubmed_title":["Comprehensive proteomics analysis of bovine sperm head plasma membrane associated with fertility."],"pubmed_authors":["Imran Muhammad M, Buhr Mary M MM, Chumala Paulos P, Katselis George S GS"],"additional_accession":[]},"is_claimable":false,"name":"Comprehensive Proteomics Analysis of Bovine Sperm Head Plasma Membrane Associated with Fertility","description":"Bull fertility impacts herd fertility, but accurately predicting male fertility from sperm characteristics is difficult once extremes are removed. The objectives of this study are identification, relative quantification, and comparison of sperm head plasma membrane (HPM) proteomics in bulls of differing bull fertility index (BFI). HPM from one fresh ejaculate from 16 Holstein bulls (8 each high and low fertility; HF, LF) was extracted and its proteins were assessed by tandem mass spectrometry. Mass spectra were aligned to UniProtKB mammal database and proteins were identified and characterized by SpectrumMill. Of 22,117 total proteins identified (unique plus homologous; 1% false discovery rate), Mass Profiler Professional statistical analysis identified 67 differentially abundant proteins (DAPs) in the HPM of 3 bulls each with highest and lowest BFI (105.66±0.54, 91.33±1.44, respectively; p<0.01]. DAPs differed in intensity by at least 2-fold (48 up-regulated, 19 down-regulated, HF vs LF) and meta-analysis confirmed their association to BFI. Gene ontology assigned up-regulated proteins to sperm fertilization mechanisms, and down-regulated proteins to catalytic and transporter activity. Linear regression positively correlated BFI of the 6 HF and LF bulls to all up-regulated DAPs (r2=0.65 to 0.97, p≤0.05), and negatively correlated BFI to all down-regulated DAPs (r2=0.76 to 0.96, p<0.05). In the 16-bull population, linear regression positively correlated BFI to 38 of the up-regulated DAPs (r2=0.29 to 0.66; p≤0.05), and negatively correlated 6 of the down-regulated DAPs (r2=0.26 to 0.44; p≤0.05). In summary, this study revealed proteins in HPM, highlighting their significant role in sperm fertilization and correlation with bull fertility.","dates":{"publication":"2026-06-08","submission":"2023-11-28"},"accession":"PXD047294","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:1073882","NEWT:6819","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: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: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:1310161","NEWT:77133","NEWT:145481","NEWT:1310165","NEWT:29058","NCBITaxon:79824","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: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: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: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: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: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: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:6493","NEWT:10832","NEWT:6494","NEWT:6491","NEWT:507601","NEWT:520","NEWT:64657","NEWT:186441","NEWT:643680","NEWT:214092","NCBITaxon:6157","NEWT:13095","NEWT:162425","NEWT:104105","NEWT:216257","NEWT:9986","NEWT:8654","NEWT:8658","NEW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