Panel of monoclonal antibodies to sperm surface proteins as a tool for monitoring localization and identification of sperm-zona pellucida receptors

0439114 - BTÚ 2016 RIV DE eng J - Journal Article
Zigo, Michal - Dorosh, Andriy - Pohlová, Alžběta - Jonáková, Věra - Šulc, Miroslav - Maňásková-Postlerová, Pavla
Panel of monoclonal antibodies to sperm surface proteins as a tool for monitoring localization and identification of sperm-zona pellucida receptors.
Cell and Tissue Research. March, č. 359 (2015), s. 895-908. ISSN 0302-766X. E-ISSN 1432-0878
R&D Projects: GA ČR(CZ) GA14-05547S; GA MŠMT(CZ) ED1.1.00/02.0109; GA ČR(CZ) GAP503/12/1834
Institutional research plan: CEZ:AV0Z50520701
Institutional support: RVO:86652036 ; RVO:61388971
Keywords : zona pellucida-binding receptors * monoclonal antibodies against sperm surface proteins * sperm surface proteins * RAB-2A * lactahedrin P47
Subject RIV: CE - Biochemistry
Impact factor: 2.948, year: 2015

Primary binding of the sperm to the zona pellucida (ZP) is one of the many steps necessary for successful fertilization. Sperm bind ZP by means of membrane receptors which recognize carbohydrate moieties on ZP glycoproteins according to a well-defined sequential process. Primary binding receptors, many of which have been disclosed in various mammals, are localized throughout the acrosomal region of the sperm surface. A panel of monoclonal antibodies against proteins from the sperm surface was prepared. Antibodies were screened by immunofluorescence for protein localization and Western blotting. Proteins localized on the sperm head and simultaneously detected by Western blotting were further studied in terms of immunolocalization in reproductive tissues and fluids, binding to ZP, immunoprecipitation and sequencing. Of 17 prepared antibodies, 8 recognized proteins localized on the sperm head and also detected proteins of interest by Western blotting. Only three other antibodies recognized proteins that also coincided in binding to ZP. These three antibodies were used for immunoprecipitation, and further protein sequencing of immunoprecipitates revealed that these antibodies distinguished acrosin precursor, RAB-2A protein, and lactadherin P47. This is not the first time we have detected acrosin on the surface of ejaculated and capacitated sperm. However, to our knowledge, this is the first time RAB-2A has been detected on the sperm surface. Lactadherin P47 has already been characterized and its physiological function in reproduction has been proposed.
Permanent Link: http://hdl.handle.net/11104/0246334