title: Bovine sperm plasma membrane proteomics through biotinylation and subcellular enrichment.
abstract: A significant proportion of mammalian fertilization is mediated through the proteomic composition of the sperm surface. These protein constituents can present as biomarkers to control and regulate breeding of agricultural animals. Previous studies have addressed the bovine sperm cell apical plasma membrane (PM) proteome with nitrogen cavitation enrichment. Alternative workflows would enable to expand the compositional data more globally around the entire sperm's surface. We used a cell surface biotin-labeling in combination with differential centrifugation to enrich sperm surface proteins. Using nano-LC MS/MS, 338 proteins were confidently identified in the PM-enriched proteome. Functional categories of sperm-egg interaction, protein turnover, metabolism as well as molecular transport, spermatogenesis, and signal transduction were represented by proteins with high quantitative signal in our study. A highly significant degree of enrichment was found for transmembrane and PM-targeted proteins. Among them, we also report proteins previously not described on bovine sperm (CPQ, CD58, CKLF, CPVL, GLB1L3, and LPCAT2B) of which CPQ and CPVL cell surface localization was further validated. A descriptive overview of the bovine sperm PM integral and peripheral proteins is provided to complement future studies on animal reproduction and its relation to sperm cell surface. All MS data have been deposited in the ProteomeXchange with identifier PXD001096 (http://proteomecentral.proteomexchange.org/dataset/PXD001096).
authors.affiliation: Proteomics Core Facility, Institute of Technology, University of Tartu, Tartu, Estonia.|Competence Centre on Reproductive Medicine and Biology, Tartu, Estonia.|Department of Obstetrics and Gynaecology, University of Tartu, Tartu, Estonia.
authors.forename: Sergo
authors.initials: S
authors.lastname: Kasvandik
authors.affiliation: Competence Centre on Reproductive Medicine and Biology, Tartu, Estonia.
authors.forename: Gerly
authors.initials: G
authors.lastname: Sillaste
authors.affiliation: Competence Centre on Reproductive Medicine and Biology, Tartu, Estonia.|Department of Obstetrics and Gynaecology, University of Tartu, Tartu, Estonia.|Center for Biology of Integrated Systems, Tallinn University of Technology, Tallinn, Estonia.
authors.forename: Agne
authors.initials: A
authors.lastname: Velthut-Meikas
authors.affiliation: Institute of Biomedicine and Translational Medicine, University of Tartu, Tartu, Estonia.
authors.forename: Aavo-Valdur
authors.initials: AV
authors.lastname: Mikelsaar
authors.affiliation: Department of Reproductive Biology, Institute of Veterinary Medicine and Animal Sciences, Estonian University of Life Sciences, Tartu, Estonia.
authors.forename: Triin
authors.initials: T
authors.lastname: Hallap
authors.affiliation: Animal Breeders Association of Estonia, Keava, Kehtna vald, Raplamaa, Estonia.
authors.forename: Peeter
authors.initials: P
authors.lastname: Padrik
authors.affiliation: Proteomics Core Facility, Institute of Technology, University of Tartu, Tartu, Estonia.
authors.forename: Tanel
authors.initials: T
authors.lastname: Tenson
authors.affiliation: Competence Centre on Reproductive Medicine and Biology, Tartu, Estonia.|Department of Reproductive Biology, Institute of Veterinary Medicine and Animal Sciences, Estonian University of Life Sciences, Tartu, Estonia.
authors.forename: Ülle
authors.initials: Ü
authors.lastname: Jaakma
authors.affiliation: Institute of Biomedicine and Translational Medicine, University of Tartu, Tartu, Estonia.
authors.forename: Sulev
authors.initials: S
authors.lastname: Kõks
authors.affiliation: Competence Centre on Reproductive Medicine and Biology, Tartu, Estonia.|Department of Obstetrics and Gynaecology, University of Tartu, Tartu, Estonia.|Institute of Biomedicine and Translational Medicine, University of Tartu, Tartu, Estonia.
authors.forename: Andres
authors.initials: A
authors.lastname: Salumets
mesh_terms: D020816:Amino Acid Motifs; D000818:Animals; D019921:Biotinylation; D002417:Cattle; D002462:Cell Membrane / Q000737:chemistry* / Q000378:metabolism; D002853:Chromatography, Liquid; D008297:Male; D008565:Membrane Proteins / Q000032:analysis* / Q000145:classification / Q000302:isolation & purification / Q000378:metabolism; D020543:Proteome / Q000302:isolation & purification; D040901:Proteomics / Q000379:methods*; D015203:Reproducibility of Results; D013094:Spermatozoa / Q000737:chemistry* / Q000166:cytology; D053719:Tandem Mass Spectrometry
chemical_list: D008565:Membrane Proteins; D020543:Proteome
keywords: Animal proteomics; Biotinylation; Bos taurus; Plasma membrane proteome; Reproductive biology; Sperm cell