{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,2,10]],"date-time":"2026-02-10T17:50:40Z","timestamp":1770745840171,"version":"3.49.0"},"reference-count":86,"publisher":"MDPI AG","issue":"4","license":[{"start":{"date-parts":[[2025,2,8]],"date-time":"2025-02-08T00:00:00Z","timestamp":1738972800000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"EPIC-XS","award":["0000186"],"award-info":[{"award-number":["0000186"]}]},{"name":"EPIC-XS","award":["EXPL\/CVT-CVT\/1112\/2021"],"award-info":[{"award-number":["EXPL\/CVT-CVT\/1112\/2021"]}]},{"name":"EPIC-XS","award":["SFRH\/BD\/146867\/2019"],"award-info":[{"award-number":["SFRH\/BD\/146867\/2019"]}]},{"name":"EPIC-XS","award":["UIDB\/04033\/2020"],"award-info":[{"award-number":["UIDB\/04033\/2020"]}]},{"name":"EPIC-XS","award":["UIDB\/CVT\/00772\/2020"],"award-info":[{"award-number":["UIDB\/CVT\/00772\/2020"]}]},{"name":"EPIC-XS","award":["UIDB\/04501\/2020"],"award-info":[{"award-number":["UIDB\/04501\/2020"]}]},{"name":"EPIC-XS","award":["UIDP\/04501\/2020"],"award-info":[{"award-number":["UIDP\/04501\/2020"]}]},{"DOI":"10.13039\/501100001871","name":"FCT\u2014Foundation for Science and Technology","doi-asserted-by":"publisher","award":["0000186"],"award-info":[{"award-number":["0000186"]}],"id":[{"id":"10.13039\/501100001871","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100001871","name":"FCT\u2014Foundation for Science and Technology","doi-asserted-by":"publisher","award":["EXPL\/CVT-CVT\/1112\/2021"],"award-info":[{"award-number":["EXPL\/CVT-CVT\/1112\/2021"]}],"id":[{"id":"10.13039\/501100001871","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100001871","name":"FCT\u2014Foundation for Science and Technology","doi-asserted-by":"publisher","award":["SFRH\/BD\/146867\/2019"],"award-info":[{"award-number":["SFRH\/BD\/146867\/2019"]}],"id":[{"id":"10.13039\/501100001871","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100001871","name":"FCT\u2014Foundation for Science and Technology","doi-asserted-by":"publisher","award":["UIDB\/04033\/2020"],"award-info":[{"award-number":["UIDB\/04033\/2020"]}],"id":[{"id":"10.13039\/501100001871","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100001871","name":"FCT\u2014Foundation for Science and Technology","doi-asserted-by":"publisher","award":["UIDB\/CVT\/00772\/2020"],"award-info":[{"award-number":["UIDB\/CVT\/00772\/2020"]}],"id":[{"id":"10.13039\/501100001871","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100001871","name":"FCT\u2014Foundation for Science and Technology","doi-asserted-by":"publisher","award":["UIDB\/04501\/2020"],"award-info":[{"award-number":["UIDB\/04501\/2020"]}],"id":[{"id":"10.13039\/501100001871","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100001871","name":"FCT\u2014Foundation for Science and Technology","doi-asserted-by":"publisher","award":["UIDP\/04501\/2020"],"award-info":[{"award-number":["UIDP\/04501\/2020"]}],"id":[{"id":"10.13039\/501100001871","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Animals"],"abstract":"<jats:p>Current sperm sexing methods are costly and largely restricted to cattle, while immunological techniques targeting sex-specific membrane proteins may offer more economical alternatives. To advance these methods, understanding the proteomic differences between the cell membranes of X- and Y-chromosome-bearing spermatozoa is essential. This study aimed to characterize the cell surface proteome of bovine sperm and identify potential targets for sperm sexing through LC-MS\/MS analysis. Cell surface protein lysates were extracted from unsexed, X-sperm (BX), and Y-sperm (BY) samples via biotinylation. Promising targets were identified through functional annotation (UniProt, eggNOG-mapper v.2.1.7) and topology prediction (DeepTMHMM v.1.0.13). Additionally, statistical overrepresentation (PANTHER 18.0) and orthology analyses were performed. Excluding contaminants, 130 proteins were detected, of which 64 proteins were detected in the BX samples and not in the BY samples. Of these, five transmembrane proteins stood out as potential X-sperm targets (ADAM2, ATP11C, DG1, MCT1, and PMCA4). They were identified as potential cell surface targets, based on GO terms and topology predictions, detected in at least two replicates of the BX samples, and shown to share orthology with other livestock species. These findings enhance our understanding of bovine sperm proteomics; however, further validation is required to confirm the utility of these five proteins in sperm sexing technologies.<\/jats:p>","DOI":"10.3390\/ani15040484","type":"journal-article","created":{"date-parts":[[2025,2,10]],"date-time":"2025-02-10T09:29:26Z","timestamp":1739179766000},"page":"484","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":2,"title":["The Surface Proteome of Bovine Unsexed and Sexed Spermatozoa"],"prefix":"10.3390","volume":"15","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-1510-4297","authenticated-orcid":false,"given":"Patr\u00edcia","family":"Pinto-Pinho","sequence":"first","affiliation":[{"name":"Centre for the Research and Technology of Agro-Environmental and Biological Sciences, University of Tr\u00e1s-os-Montes and Alto Douro, 5000-801 Vila Real, Portugal"},{"name":"Laboratory of Signal Transduction, Department of Medical Sciences, Institute of Biomedicine, University of Aveiro, 3810-193 Aveiro, Portugal"},{"name":"Laboratory of Genetics and Andrology, Hospital Center of Tr\u00e1s-os-Montes and Alto Douro, E.P.E., 5000-508 Vila Real, Portugal"},{"name":"Experimental Pathology and Therapeutics Group, IPO Porto Research Center, Portuguese Institute of Oncology of Porto Francisco Gentil, E.P.E., 4200-072 Porto, Portugal"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-2039-0360","authenticated-orcid":false,"given":"Joana","family":"Quelhas","sequence":"additional","affiliation":[{"name":"Laboratory of Signal Transduction, Department of Medical Sciences, Institute of Biomedicine, University of Aveiro, 3810-193 Aveiro, Portugal"},{"name":"Department of Veterinary Clinics, Institute of Biomedical Sciences Abel Salazar, University of Porto, 4050-313 Porto, Portugal"},{"name":"Department of Veterinary Sciences, University School Vasco da Gama, 3020-210 Coimbra, Portugal"}]},{"given":"Francis","family":"Impens","sequence":"additional","affiliation":[{"name":"VIB Proteomics Core, B-9052 Ghent, Belgium"},{"name":"VIB-UGent Center for Medical Biotechnology, Flemish Institute for Biotechnology, B-9052 Ghent, Belgium"},{"name":"Department of Biomolecular Medicine, Ghent University, B-9052 Ghent, Belgium"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-2036-7849","authenticated-orcid":false,"given":"Sara","family":"Dufour","sequence":"additional","affiliation":[{"name":"VIB Proteomics Core, B-9052 Ghent, Belgium"},{"name":"VIB-UGent Center for Medical Biotechnology, Flemish Institute for Biotechnology, B-9052 Ghent, Belgium"},{"name":"Department of Biomolecular Medicine, Ghent University, B-9052 Ghent, Belgium"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-4659-5631","authenticated-orcid":false,"given":"Delphi","family":"Van Haver","sequence":"additional","affiliation":[{"name":"VIB Proteomics Core, B-9052 Ghent, Belgium"},{"name":"VIB-UGent Center for Medical Biotechnology, Flemish Institute for Biotechnology, B-9052 Ghent, Belgium"},{"name":"Department of Biomolecular Medicine, Ghent University, B-9052 Ghent, Belgium"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-6734-974X","authenticated-orcid":false,"given":"Gra\u00e7a","family":"Lopes","sequence":"additional","affiliation":[{"name":"Department of Veterinary Clinics, Institute of Biomedical Sciences Abel Salazar, University of Porto, 4050-313 Porto, Portugal"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-9710-9835","authenticated-orcid":false,"given":"Ant\u00f3nio","family":"Rocha","sequence":"additional","affiliation":[{"name":"Department of Imuno-Physiology and Pharmacology, Institute of Biomedical Sciences Abel Salazar, University of Porto, 4050-313 Porto, Portugal"}]},{"given":"Ros\u00e1rio","family":"Pinto-Leite","sequence":"additional","affiliation":[{"name":"Laboratory of Genetics and Andrology, Hospital Center of Tr\u00e1s-os-Montes and Alto Douro, E.P.E., 5000-508 Vila Real, Portugal"},{"name":"Experimental Pathology and Therapeutics Group, IPO Porto Research Center, Portuguese Institute of Oncology of Porto Francisco Gentil, E.P.E., 4200-072 Porto, Portugal"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-7459-9173","authenticated-orcid":false,"given":"Margarida","family":"Fardilha","sequence":"additional","affiliation":[{"name":"Laboratory of Signal Transduction, Department of Medical Sciences, Institute of Biomedicine, University of Aveiro, 3810-193 Aveiro, Portugal"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-4879-8624","authenticated-orcid":false,"given":"Bruno","family":"Cola\u00e7o","sequence":"additional","affiliation":[{"name":"Animal and Veterinary Research Centre, University of Tr\u00e1s-os-Montes and Alto Douro, 5001-801 Vila Real, Portugal"}]}],"member":"1968","published-online":{"date-parts":[[2025,2,8]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"s97","DOI":"10.1017\/S1751731118000721","article-title":"Review: 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