title: Albumin is synthesized in epididymis and aggregates in a high molecular mass glycoprotein complex involved in sperm-egg fertilization.
abstract: The epididymis has an important role in the maturation of sperm for fertilization, but little is known about the epididymal molecules involved in sperm modifications during this process. We have previously described the expression pattern for an antigen in epididymal epithelial cells that reacts with the monoclonal antibody (mAb) TRA 54. Immunohistochemical and immunoblotting analyses suggest that the epitope of the epididymal antigen probably involves a sugar moiety that is released into the epididymal lumen in an androgen-dependent manner and subsequently binds to luminal sperm. Using column chromatography, SDS-PAGE with in situ digestion and mass spectrometry, we have identified the protein recognized by mAb TRA 54 in mouse epididymal epithelial cells. The ∼65 kDa protein is part of a high molecular mass complex (∼260 kDa) that is also present in the sperm acrosomal vesicle and is completely released after the acrosomal reaction. The amino acid sequence of the protein corresponded to that of albumin. Immunoprecipitates with anti-albumin antibody contained the antigen recognized by mAb TRA 54, indicating that the epididymal molecule recognized by mAb TRA 54 is albumin. RT-PCR detected albumin mRNA in the epididymis and fertilization assays in vitro showed that the glycoprotein complex containing albumin was involved in the ability of sperm to recognize and penetrate the egg zona pellucida. Together, these results indicate that epididymal-derived albumin participates in the formation of a high molecular mass glycoprotein complex that has an important role in egg fertilization.
authors.affiliation: Department of Histology and Embryology, Institute of Biology, State University of Campinas (UNICAMP), Campinas, São Paulo, Brazil.
authors.forename: Kélen Fabíola
authors.initials: KF
authors.lastname: Arroteia
authors.affiliation: Department of Histology and Embryology, Institute of Biology, State University of Campinas (UNICAMP), Campinas, São Paulo, Brazil.
authors.forename: Mainara Ferreira
authors.initials: MF
authors.lastname: Barbieri
authors.affiliation: Department of Pharmacology, Faculty of Medical Sciences, State University of Campinas (UNICAMP), Campinas, São Paulo, Brazil; ThoMSon Mass Spectrometry Laboratory, Department of Organic Chemistry, Institute of Chemistry, State University of Campinas (UNICAMP), Campinas, São Paulo, Brazil.
authors.forename: Gustavo Henrique Martins Ferreira
authors.initials: GH
authors.lastname: Souza
authors.affiliation: Faculty of Pharmaceutical Sciences, Nagasaki International University, Nagasaki, Japan.
authors.forename: Hiromitsu
authors.initials: H
authors.lastname: Tanaka
authors.affiliation: ThoMSon Mass Spectrometry Laboratory, Department of Organic Chemistry, Institute of Chemistry, State University of Campinas (UNICAMP), Campinas, São Paulo, Brazil.
authors.forename: Marcos Nogueira
authors.initials: MN
authors.lastname: Eberlin
authors.affiliation: Department of Pharmacology, Faculty of Medical Sciences, State University of Campinas (UNICAMP), Campinas, São Paulo, Brazil.
authors.forename: Stephen
authors.initials: S
authors.lastname: Hyslop
authors.affiliation: Department of Histology and Embryology, Institute of Biology, State University of Campinas (UNICAMP), Campinas, São Paulo, Brazil.
authors.forename: Lúcia Elvira
authors.initials: LE
authors.lastname: Alvares
authors.affiliation: Department of Histology and Embryology, Institute of Biology, State University of Campinas (UNICAMP), Campinas, São Paulo, Brazil.
authors.forename: Luís Antonio Violin Dias
authors.initials: LA
authors.lastname: Pereira
mesh_terms: D000418:Albumins / Q000378:metabolism*; D000818:Animals; D000911:Antibodies, Monoclonal; D004591:Electrophoresis, Polyacrylamide Gel; D004822:Epididymis / Q000378:metabolism*; D005260:Female; D005306:Fertilization / Q000502:physiology; D006023:Glycoproteins / Q000378:metabolism*; D015151:Immunoblotting; D047468:Immunoprecipitation; D008297:Male; D051379:Mice; D008810:Mice, Inbred C57BL; D010063:Ovum / Q000378:metabolism* / Q000502:physiology; D021241:Spectrometry, Mass, Electrospray Ionization; D013094:Spermatozoa / Q000378:metabolism* / Q000502:physiology
chemical_list: D000418:Albumins; D000911:Antibodies, Monoclonal; D006023:Glycoproteins
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