Aggregated and monomeric forms of proteins in boar seminal plasma: characterization and binding properties.
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.
Manásková
P
P
Institute of Molecular Genetics, Academy of Sciences of the Czech Republic, Prague.
Liberda
J
J
Tichá
M
M
Jonáková
V
V
D000176:Acrosin / Q000037:antagonists & inhibitors; D000818:Animals; D001360:Avidin; D019921:Biotinylation; D002352:Carrier Proteins / Q000737:chemistry / Q000378:metabolism*; D002784:Cholesterol / Q000031:analogs & derivatives / Q000378:metabolism; D002809:Chondroitin Sulfates / Q000737:chemistry / Q000494:pharmacology; D002850:Chromatography, Gel; D002851:Chromatography, High Pressure Liquid; D005260:Female; D005456:Fluorescent Dyes; D006023:Glycoproteins / Q000737:chemistry / Q000378:metabolism*; D006493:Heparin / Q000031:analogs & derivatives / Q000378:metabolism; D008297:Male; D008562:Membrane Glycoproteins / Q000737:chemistry / Q000378:metabolism*; D011485:Protein Binding; D012661:Semen / Q000737:chemistry*; D029589:Seminal Plasma Proteins; D029607:Seminal Vesicle Secretory Proteins; D013084:Sperm-Ovum Interactions; D013094:Spermatozoa / Q000737:chemistry / Q000378:metabolism*; D013552:Swine; D015044:Zona Pellucida / Q000737:chemistry / Q000378:metabolism
D000176:Acrosin; D001360:Avidin; D002352:Carrier Proteins; D002784:Cholesterol; D002809:Chondroitin Sulfates; D005456:Fluorescent Dyes; D006023:Glycoproteins; D006493:Heparin; D008562:Membrane Glycoproteins; D029589:Seminal Plasma Proteins; D029607:Seminal Vesicle Secretory Proteins; C121098:BSP1 protein, Sus scrofa; C425042:spermadhesin; C074520:seminal vesicle secretory protein 109, porcine