Boar seminal plasma was separated into five protein fractions (I-V) (>100, 55, 45, 30, 5-15 kDa) by gel filtration chromatography on Sephadex G-75 SF at pH 7.4. RP HPLC of protein fractions I-V and N-terminal sequencing of their individual components revealed that high-molecular-weight aggregates consisted mainly of DQH sperm surface protein and AQN, AWN, PSP II spermadhesins, while fraction IV consisted of heterodimers of PSP spermadhesins only. Spermadhesins as monomers were present in seminal plasma in a very low amount. Biotinylated fractions I-IV containing AWN, AQN, DQH, and PSP proteins were bound to boar epididymal and ejaculated spermatozoa with the same efficiency. Aggregates containing AWN, AQN, DQH, PSP II proteins (fractions I-III) and their HPLC-separated monomeric forms interacted with phosphorylcholine. Aggregates containing the DQH protein and AWN spermadhesins as well as their separated monomeric proteins interacted strongly with acidic polysaccharides. PSP II interacted with some acidic polysaccharides, while the fraction IV corresponding to heterodimer PSP IPSP II did not show any binding to acidic polysaccharides and zona pellucida. Fractions I-III showed affinity to cholesterol. The strongest interaction was observed between biotinylated glycoproteins of porcine zona pellucida and AWN 1-containing aggregates and separated proteins. AQN 1 spermadhesin effectively blocked the sperm binding to oocytes. These results suggest that under physiological conditions, the aggregated forms of seminal plasma proteins (DQH, AQN, AWN, PSP II) rather than the individual proteins might take part in coating the sperm surface, in sperm capacitation and in primary binding of spermatozoa to zona pellucida of the ovum.

• MeSH
• akrosin antagonisté a inhibitory   MeSH
• avidin MeSH
• biotinylace MeSH
• cholesterol analogy a deriváty   metabolismus   MeSH
• chondroitinsulfáty chemie   farmakologie   MeSH
• fluorescenční barviva MeSH
• gelová chromatografie MeSH
• glykoproteiny chemie   metabolismus   MeSH
• heparin analogy a deriváty   metabolismus   MeSH
• interakce spermie a vajíčka MeSH
• membránové glykoproteiny chemie   metabolismus   MeSH
• prasata * MeSH
• proteiny semenné plazmy * MeSH
• sekreční proteiny semenných váčků * MeSH
• sperma chemie   MeSH
• spermie chemie   metabolismus   MeSH
• transportní proteiny chemie   metabolismus   MeSH
• vazba proteinů MeSH
• vysokoúčinná kapalinová chromatografie MeSH
• zona pellucida chemie   metabolismus   MeSH
• zvířata MeSH
• Check Tag
• mužské pohlaví MeSH
• ženské pohlaví MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• akrosin MeSH
• avidin MeSH
• cholesterol MeSH
• chondroitinsulfáty MeSH
• DQH sperm surface protein, Sus scrofa MeSH Prohlížeč
• fluorescenční barviva MeSH
• glykoproteiny MeSH
• heparin MeSH
• membránové glykoproteiny MeSH
• proteiny semenné plazmy * MeSH
• sekreční proteiny semenných váčků * MeSH
• seminal vesicle secretory protein 109, porcine MeSH Prohlížeč
• spermadhesin MeSH Prohlížeč
• transportní proteiny MeSH

Boar seminal plasma proteins were separated by gel chromatography on Sephadex G-75 into five fractions (I-V). Serine proteinase inhibitors were found mainly in the protein fraction with relative molecular weight 5-25kDa. Small amounts of these inhibitors were also found in the high molecular weight protein fraction (M(r)>100kDa). The protein fraction containing most of the proteinase inhibitory activity was further separated by RP HPLC. Isolated proteins were characterized by SDS electrophoresis and immunoblotting, N-terminal amino acid sequencing and by determination of the proteinase inhibitory activity. In the fraction containing proteinase inhibitors, also beta-microseminoprotein (beta-MSP), AQN 1 and lactoferrin were identified. The possible existence of complexes of protein components in the fraction with relative molecular weight 5-25kDa was studied in detail using gel chromatographic separation on Sephadex G-50. A part of proteinase inhibitors with M(r) 8kDa was eluted together with AQN 1 spermadhesin. An interaction of isolated spermadhesin AQN 1 and proteinase inhibitor was shown.

• MeSH
• gelová chromatografie MeSH
• inhibitory proteas chemie   izolace a purifikace   farmakologie   MeSH
• molekulární sekvence - údaje MeSH
• molekulová hmotnost MeSH
• sekvence aminokyselin MeSH
• sperma chemie   MeSH
• Sus scrofa * MeSH
• vysokoúčinná kapalinová chromatografie MeSH
• zvířata MeSH
• Check Tag
• mužské pohlaví MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• inhibitory proteas MeSH

Boar seminal plasma proteins were separated by affinity chromatography on immobilized heparin into two portions: heparin-binding (H+) and non-heparin-binding (H-) proteins. Gel chromatography of the H+ portion yielded four main protein fractions of >150, 45, 30 and 20 kDa, while that of the H- portion resulted in the separation into three main protein fractions of >150, 30 and 20 kDa. HPLC analysis and N-terminal sequencing used to characterize the composition of the protein fractions obtained by gel chromatography revealed that all consisted of low (12-16 kDa) molecular weight components: the H+ fraction consisted of DQH sperm protein, AQN and AWN spermadhesins whereas the H- fraction consisted of PSPI and PSPII spermadhesins. The high molecular weight values of fractions obtained by gel chromatography thus suggest that the proteins are present in boar seminal plasma in the form of aggregates. Interactions of individual boar seminal plasma proteins and their aggregates present in the H+ and H- fractions with acid polysaccharides were estimated.

• MeSH
• chondroitinsulfáty metabolismus   MeSH
• chromatografie afinitní MeSH
• heparin metabolismus   MeSH
• kapacitace spermií MeSH
• kyselina hyaluronová metabolismus   MeSH
• makromolekulární látky MeSH
• molekulová hmotnost MeSH
• polysacharidy metabolismus   MeSH
• prasata metabolismus   MeSH
• proteiny izolace a purifikace   metabolismus   MeSH
• sekvenční analýza proteinů MeSH
• síran dextranu metabolismus   MeSH
• sperma chemie   MeSH
• tělesné tekutiny chemie   MeSH
• vazba proteinů MeSH
• vysokoúčinná kapalinová chromatografie MeSH
• zona pellucida metabolismus   MeSH
• zvířata MeSH
• Check Tag
• mužské pohlaví MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• srovnávací studie MeSH
• Názvy látek
• chondroitinsulfáty MeSH
• fucoidan MeSH Prohlížeč
• heparin MeSH
• kyselina hyaluronová MeSH
• makromolekulární látky MeSH
• polysacharidy MeSH
• proteiny MeSH
• síran dextranu MeSH