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Panel of monoclonal antibodies to sperm surface proteins as a tool for monitoring localization and identification of sperm-zona pellucida receptors
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SYSNO ASEP 0439114 Document Type J - Journal Article R&D Document Type Journal Article Subsidiary J Článek ve WOS Title Panel of monoclonal antibodies to sperm surface proteins as a tool for monitoring localization and identification of sperm-zona pellucida receptors Author(s) Zigo, Michal (BTO-N) RID
Dorosh, Andriy (BTO-N) RID
Pohlová, Alžběta (BTO-N)
Jonáková, Věra (BTO-N) RID
Šulc, Miroslav (MBU-M) RID, ORCID
Maňásková-Postlerová, Pavla (BTO-N) ORCIDNumber of authors 6 Source Title Cell and Tissue Research - ISSN 0302-766X
March, č. 359 (2015), s. 895-908Number of pages 14 s. Language eng - English Country DE - Germany Keywords zona pellucida-binding receptors ; monoclonal antibodies against sperm surface proteins ; sperm surface proteins ; RAB-2A ; lactahedrin P47 Subject RIV CE - Biochemistry R&D Projects GA14-05547S GA ČR - Czech Science Foundation (CSF) ED1.1.00/02.0109 GA MŠMT - Ministry of Education, Youth and Sports (MEYS) GAP503/12/1834 GA ČR - Czech Science Foundation (CSF) Institutional support BTO-N - RVO:86652036 ; MBU-M - RVO:61388971 CEZ AV0Z50520701 - BTO-N (2007-2013) UT WOS 000350351400015 DOI 10.1007/s00441-014-2072-9 Annotation 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. Workplace Institute of Biotechnology Contact Monika Kopřivová, Monika.Koprivova@ibt.cas.cz, Tel.: 325 873 700 Year of Publishing 2016
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