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Zinc-binding proteins from boar seminal plasma - isolation, biochemical characteristics and influence on spermatozoa stored at 4°C

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Mogielnicka-Brzozowska M., Wysocki P., Strzeżek J., Kordan W.
Acta Biochimica Polonica
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2011
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vol. 58
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issue 2
171-177
EN
Affinity chromatography on Chelating Sepharose Fast Flow Gel-Zn2+ was used for fractionation of boar seminal plasma proteins. Approximately 30% of total boar seminal plasma proteins showed affinity for zinc ions (ZnBP fraction). Native electrophoresis (PAGE) of ZnBP revealed six protein fractions which separated into 27 bands under denaturing conditions (SDS/PAGE). Two-dimensional electrophoresis (2D PAGE) showed 148 polypeptides with isoelectric points mostly in the basic and neutral pH range. The zinc-binding proteins comprise mainly 10-20 kDa polypeptides which are probably members of the spermadhesin family. ZnBP present in the incubation mixture of spermatozoa stored for 1 or 24 h at 4 °C allowed preservation of a higher percentage of cells exhibiting linear motility in comparison to a control sample stored in PBS. Presented results indicate that proteins binding Zn2+ ions have a shielding effect on the sperm plasma membrane and acrosome of spermatozoa, protecting these structures against consequences of cold shock.
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The antigenic character of zinc-binding proteins and their localization in boar reproductive tract - A preliminary study

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Mogielnicka-Brzozowska M., Fraser L., Strzeżek J., Kordan W.
Acta Biochimica Polonica
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2013
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vol. 60
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issue 2
255-258
EN
In this study immunoelectrophoretic and double immunodiffusion analyses were used to investigate the antigenic character of zinc-binding proteins (ZnBPs), whereas the indirect immunofluorescence technique was used to identify their origin in boar reproductive tract. The mmunoelectrophoretic analysis of ZnBPs of the seminal plasma resulted in the appearance of three antigenic protein complexes, while specific immunoreactivity patterns of the anti-ZnBP serum were detected by double immunodiffusion analysis. Indirect immunofluorescence technique confirmed that ZnBPs were secreted by different reproductive tract tissues, suggesting their contributions to the seminal plasma.
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