Biochemical characterization of CD52-like molecule in bull sperm and boar seminal plasma.\n\n

0474835 - BTÚ 2018 CZ eng A - Abstrakt
Michalková, K. - Simon, M. - Antalíková, J. - Jankovičová, J. - Šecová, P. - Horovská, L. - Postlerová, Pavla
Biochemical characterization of CD52-like molecule in bull sperm and boar seminal plasma.

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Book of abstracts XXIIIrd Symposium of Immunology and Biology of Reproduction. Vestec u Prahy: Biotechnologický ústav AVČR v. v. i., 2017 - (Kubátová, a.). s. 17-17
[XXIIIrd Symposium of Immunology and Biology of Reproduction. 18.05.2017-20.05.2017, Třešť]
Grant CEP: GA MŠMT(CZ) ED1.1.00/02.0109; GA AV ČR(CZ) SAV-AVČR 15-05
Institucionální podpora: RVO:86652036
Klíčová slova: CD52-like molecule * bull sperm * boar sperm * seminal plasma proteins * monoclonal antibody
Obor OECD: Biochemistry and molecular biology

The aim of this study was to analyse the biochemical properties of the CD52-like antigen in bull sperm and boar seminal plasma using monoclonal antibody (mAb) IVA-543 prepared in the laboratory at CBs SAS. Human CD52 is a GPI-anchored protein with molecular weight in the range from 25 to 29 kDa, which is expressed on all lymphocytes and in the male genital tract. Except human, CD52 has been described in mice, rats, chimps, dogs and pigs. In all species, CD52 is characterised by several common features, such as a post-testicular production in the male genital tract, similar expression in epididymal epithelium and sperm membrane distribution. CD52 molecule is also considered as a major sperm maturation antigen. Western blot analysis showed that the molecular weight of bull CD52-like antigen in sperm extract is 18-22 kDa whereas in seminal plasma 16-20 kDa. Two-dimensional electrophoresis of bull sperm extract and boar seminal plasma showed pI of CD52-like protein in the range from 3.1 to 3.6 and 4.5 to 5.5, respectively. Molecular mass of boar seminal plasma CD52-like appeared as 20-28 kDa. Neuraminidase treatment of bull sperm extract caused 2 kDa reduction of CD52-like molecular weight, which suggests sialic acid modification. However, presumably glycosylation of the antigen was not confirmed after glycopeptidase F and O-glycosidase treatments, since molecular weight of the protein was unchanged. Treatment of bull sperm with phosphatidyl-inositol specific phospholipase C (PI-PLC) suggested that antigen is GPI-anchored, because the ratio of mAb IVA-543 reactive sperm decreased by 70%. This findings was also supported by immunoprecipitation of bull antigen remained in supernatant after the PI-PLC treatment of sperm. Molecular weight of immunoprecipitated protein was 20 kDa. Nevertheless, the protein in bull sperm and boar seminal plasma recognized by our mAb IVA-543 is required for the CD52 molecule confirmation by mass spectrometry.
Trvalý link: http://hdl.handle.net/11104/0276839