{"database":"Pride","file_versions":[{"headers":{"Content-Type":["application/json"]},"body":{"files":{"Txt":["ftp://ftp.pride.ebi.ac.uk/pride/data/archive/2026/04/PXD065290/Sperm-MOV-proteinGroups.txt","ftp://ftp.pride.ebi.ac.uk/pride/data/archive/2026/04/PXD065290/quant_evidence.txt","ftp://ftp.pride.ebi.ac.uk/pride/data/archive/2026/04/PXD065290/Sperm-MOV-evidence.txt","ftp://ftp.pride.ebi.ac.uk/pride/data/archive/2026/04/PXD065290/Sperm-ARI-evidence.txt","ftp://ftp.pride.ebi.ac.uk/pride/data/archive/2026/04/PXD065290/Sperm-ARI-proteinGroups.txt","ftp://ftp.pride.ebi.ac.uk/pride/data/archive/2026/04/PXD065290/Sperm-ARI-parameters.txt","ftp://ftp.pride.ebi.ac.uk/pride/data/archive/2026/04/PXD065290/Sperm-MOV-parameters.txt","ftp://ftp.pride.ebi.ac.uk/pride/data/archive/2026/04/PXD065290/quant_parameters.txt","ftp://ftp.pride.ebi.ac.uk/pride/data/archive/2026/04/PXD065290/quant_proteinGroups.txt"],"Raw":["ftp://ftp.pride.ebi.ac.uk/pride/data/archive/2026/04/PXD065290/SILAC-SFP-CCE20200905-rep6-hfx2.raw","ftp://ftp.pride.ebi.ac.uk/pride/data/archive/2026/04/PXD065290/SILAC-SFP-CCE20200904-rep2-hfx2.raw","ftp://ftp.pride.ebi.ac.uk/pride/data/archive/2026/04/PXD065290/SILAC-SFP-CCE20200905-rep5-hfx1.raw","ftp://ftp.pride.ebi.ac.uk/pride/data/archive/2026/04/PXD065290/SILAC-SFP-CCE20200904-rep1-hfx1.raw","ftp://ftp.pride.ebi.ac.uk/pride/data/archive/2026/04/PXD065290/SILAC-SFP-CCE20200905-rep5-hfx2.raw","ftp://ftp.pride.ebi.ac.uk/pride/data/archive/2026/04/PXD065290/SILAC-SFP-CCE20200904-rep1-hfx2.raw","ftp://ftp.pride.ebi.ac.uk/pride/data/archive/2026/04/PXD065290/SILAC-SFP-CCE20200905-rep4-hfx1.raw","ftp://ftp.pride.ebi.ac.uk/pride/data/archive/2026/04/PXD065290/MOV-sperm.raw","ftp://ftp.pride.ebi.ac.uk/pride/data/archive/2026/04/PXD065290/SILAC-SFP-CCE20200904-rep3-hfx1.raw","ftp://ftp.pride.ebi.ac.uk/pride/data/archive/2026/04/PXD065290/SILAC-SFP-CCE20200904-rep3-hfx2.raw","ftp://ftp.pride.ebi.ac.uk/pride/data/archive/2026/04/PXD065290/SILAC-SFP-CCE20200905-rep4-hfx2.raw","ftp://ftp.pride.ebi.ac.uk/pride/data/archive/2026/04/PXD065290/SILAC-SFP-CCE20200904-rep2-hfx1.raw","ftp://ftp.pride.ebi.ac.uk/pride/data/archive/2026/04/PXD065290/ARI-sperm.raw","ftp://ftp.pride.ebi.ac.uk/pride/data/archive/2026/04/PXD065290/SILAC-SFP-CCE20200905-rep6-hfx1.raw"],"Fasta":["ftp://ftp.pride.ebi.ac.uk/pride/data/archive/2026/04/PXD065290/ARIcat10genename.fasta","ftp://ftp.pride.ebi.ac.uk/pride/data/archive/2026/04/PXD065290/MOVcat10genename.fasta"],"Mztab":["ftp://ftp.pride.ebi.ac.uk/pride/data/archive/2026/04/PXD065290/Sperm-MOV-mzTab.mzTab","ftp://ftp.pride.ebi.ac.uk/pride/data/archive/2026/04/PXD065290/Sperm-ARI-mzTab.mzTab","ftp://ftp.pride.ebi.ac.uk/pride/data/archive/2026/04/PXD065290/SILAC-SFP-mzTab.mzTab"]},"type":"primary"},"statusCode":"OK","statusCodeValue":200}],"scores":null,"additional":{"labhead_mail":["lmatzkin@arizona.edu"],"submitter":["Luciano Matzkin"],"technology_type":["Data-dependent acquisition","Mass Spectrometry"],"software":[""],"submitter_keywords":[""],"full_dataset_link":["https://www.ebi.ac.uk/pride/archive/projects/PXD065290"],"tissue":["Female Reproductive System","Sperm","Diploid Cell"],"sample_protocol":["Mated lower female reproductive tracts of D. mojavensis and D. arizonae were denatured, reduced and alkylated with 5% (w/v) sodium dodecyl sulfate (SDS), 10mM tris (2-carboxyethyl) phosphine hydrochloride (TCEP-HCl), 40mM 2-chloroacetamide, 50mM Tris pH 8.5 and boiled at 95°C for 10 minutes. The SP3 method[39] was utilized to prepare samples for mass spectrometry analyses. Protein lysates were treated with carboxylate-functionalized speedbeads (Cytiva) and acetonitrile was added to 80% (v/v) to promote binding to the beads. Beads were then washed twice with 80% (v/v) ethanol and twice with 100% acetonitrile. Digestion was performed with LysC/Trypsin mix (Promega) at a 1:50 protease to protein ratio in 50mM Tris pH 8.5. Samples were incubated and rotated at 37°C overnight. Digests were purified with an Oasis HLB 1cc (10mg) cartridge (Waters) following the manufacturers protocol and then dried using a vacuum rotatory evaporator. Peptides were directly injected onto a Waters M-class column (1.7 m, 120A, rpC18, 75 m x 250mm) and eluted from 2% to 20% acetonitrile with 0.1%(v/v) formic acid for 100 minutes then 20% to 32% acetonitrile for 20 minutes at 0.3 l/minute using a Thermo Ultimate 3000 UPLC (Thermo Scientific). Mass spectrometry was performed with a Thermo Q-Exactive HF-X mass spectrometer (Thermo Scientific). MS1 scans were performed at 120,000 resolution from 380 to 1580 m/z with a 45ms fill time and 3E6 AGC target. For MS2 scans, the 12 most intense peaks were isolated with 1.4 m/z window with 100ms fill time and 1E5 AGC target and 27% HCD collision energy at 15,000 resolution. Dynamic exclusion was enabled for 25 seconds."],"repository":["Pride"],"quantification_method":[""],"modification":[""],"data_protocol":["Proteins were identified and quantified using MaxQuant, assuming a triple SILAC labeling experiment (heavy [Lys8] =D. mojavensis SFPs, medium [Lys4] =D. arizonae SFPs, light [Lys] =D. mojavensis and D. arizonae female proteins). The protein database comprised combined D. mojavensis (NDSSC: 15081-1353.01) and D. arizonae (NDSSC: 15081-1271.41) proteins using the longest transcript for each species (cactusflybase R2024_09). The analysis included the possibility of common variable modifications (Oxidation (M); Acetyl (Protein N-term) and PSM and protein FDR were set to 0.01. We required at least two peptides for a protein to be identified. Because variation in ionization efficiencies of different peptides can reduce the reliability of abundance estimates, we filtered our SFP dataset for quantitative comparisons to include only peptides shared between the species."],"omics_type":["Proteomics"],"labhead":["Luciano Matzkin"],"instrument_platform":[""],"submission_type":["COMPLETE"],"labhead_affiliation":["Department of Entomology BIO5 Institute Department of Ecology and Evolutionary Biology University of Arizona"],"species":["Drosophila Mojavensis Mojavensis","Drosophila Arizonae"],"submitter_mail":["lmatzkin@arizona.edu"],"publication":["Not available"],"submitter_affiliation":["University of Arizona"],"submitter_country":["United States"],"doi":["10.6019/PXD065290"],"additional_accession":[]},"is_claimable":false,"name":"Evolution of the reproductive interactome and the origins of postmating–prezygotic reproductive divergence","description":"Reproductive proteins evolve rapidly, which is predicted to cause postmating-prezygotic (PMPZ) reproductive isolation between divergent populations. While most previous studies have focused on protein sequence divergence as a likely driver of PMPZ incompatibilities, reproductive proteomes may also diverge compositionally and/or quantitatively. Here, we characterize extensive protein sequence, compositional, and quantitative divergence of male and female reproductive proteomes in two species exhibiting strong PMPZ isolation (D. mojavensis and D. arizonae). We further demonstrate that three divergent male seminal fluid proteins affect the size of the insemination reaction mass and/or fertilization success in D. arizonae. Using quantitative data and protein-protein interaction modeling of proteases and protease inhibitors, we predict the relative importance of protein sequence divergence, compositional changes, and quantitative divergence to the evolution of PMPZ incompatibilities between the species. We find that predicted interspecies protein incompatibilities arise from proteome compositional changes rather than protein sequence divergence, at least for the subset of the interactome that we tested. Furthermore, extensive quantitative divergence, particularly for proteases and inhibitors, suggests pervasive stoichiometric mismatches in heterospecific matings. Altogether, our findings provide novel insights into how the reproductive interactome diverges, and the consequences of this divergence for the evolution of PMPZ reproductive isolation.","dates":{"publication":"2026-04-02","submission":"2025-06-21"},"accession":"PXD065290","cross_references":{"TAXONOMY":["NEWT:3555","NEWT:71647","NEWT:330879","NEWT:35554","NEWT:32046","NEWT:544496","NEWT:2042546","NEWT:32049","NEWT:1392696","NEWT:259447","NEWT:295546","NEWT:45351","NEWT:445974","NEWT:43179","NEWT:180454","NEWT:5722","NCBITaxon:1280","NEWT:1129","NEWT:309807","NEWT:55153","NEWT:309800","NEWT:281395","NEWT:1211601","NEWT:876138","NEWT:44271","NEWT:193516","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","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:7959","NEWT:2261","NEWT:31156","NEWT:3112","NEWT:79329","NEWT:4442","NEWT:192875","NEWT:12637","NEWT:59729","NEWT:2164133","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","NCBITaxon:4563","NEWT:5755","NEWT:44689","NEWT:44447","NEWT:752555","NEWT:498211","NEWT:5759","NEWT:1736231","NEWT:5518","NEWT:398007","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: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:2065263","NEWT:1772","NEWT:118698","NEWT:1616117","NEWT:263","NEWT:118696","NEWT:52283","NEWT:11988","NEWT:284812","NEWT:8175","NEWT:400667","NEWT:43330","NEWT:1603293","NEWT:408169","NEWT:44664","NEWT:47946","NEWT:3702","NEWT:243277","NEWT:1246791","NEWT:7067","NEWT:118694","NEWT:2850","NEWT:118691","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:376686","NEWT:95486","NEWT:508771","NEWT:1883446","NEWT:253","NCBITaxon:2759","NEWT:1233435","NEWT:93061","NEWT:93062","NCBITaxon:4932","NEWT:109760","NEWT:943274","NEWT:644223","NEWT:235443","NEWT:108458","NEWT:5936","NEWT:272623","NEWT:272624","NEWT:320637","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:1255228","NEWT:649908","NEWT:410289","NEWT:373153","NEWT:3983","NEWT:352472","NEWT:1071661","NEWT:1202532","NEWT:360094","NEWT:470","NEWT:41364","NEWT:1313","NEWT:1218097","NCBITaxon:50557","NEWT:39251","NEWT:39491","NCBITaxon:5811","NEWT:29491","NEWT:101841","NEWT:2014887","NEWT:33952","NEWT:153009","NEWT:261756","NEWT:571256","NEWT:40483","NEWT:63366","NEWT:63367","NEWT:688333","NEWT:215402","NCBITaxon:548681","NEWT:318586","NEWT:272634","NEWT:198107","NEWT:9031","NEWT:1872122","NEWT:1308","NEWT:7091","NEWT:1034015","NEWT:760568","NEWT:141262","NEWT:108931","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:5180","NEWT:256737","NEWT:4090","NEWT:4092","NEWT:9541","NEWT:8694","NEWT:4093","NEWT:4096","NEWT:8692","NEWT:2903","NEWT:185579","NEWT:941442","NEWT:536088","NEWT:13076","NEWT:1226408","NEWT:7108","NEWT:40479","NEWT:43989","NEWT:4076","NEWT:142615","NEWT:317","NEWT:1006581","NEWT:885318","NEWT:39488","NEWT:550","NEWT:4081","NEWT:273068","NEWT:554","NEWT:98334","NEWT:451516","NEWT:4084","NEWT:63577","NEWT:36185","NEWT:1225786","NEWT:7574","NEWT:1715256","NEWT:30640","NEWT:575412","NEWT:929793","NEWT:29204","NEWT:28112","NEWT:114155","NEWT:6494","NEWT:6491","NEWT:507601","NEWT:520","NEWT:186441","NEWT:643680","NEWT:214092","NCBITaxon:6157","NEWT:6239","NEWT:162425","NEWT:216257","NEWT: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