Boar spermadhesin AWN-1. Oligosaccharide and zona pellucida binding characteristics.
We have analyzed the oligosaccharide recognition ability of boar spermadhesin AWN-1 using biotinylated glycoproteins with defined carbohydrate chains as probes. Our results show that AWN-1 bound to proteins containing O-linked NeuAc alpha(2-3/6)-Gal beta(1-3)-GalNAc (PDC-109 and fetuin) with a Kd = 0.7 microM. AWN-1 also bound to NeuAc alpha(2-3/6)-Gal beta(1-4)-GlcNAc sequences in N-linked triantennary structures of fetuin, but not to the same oligosaccharide in the diantennary structures of IgG or fibrinogen. The absence of terminal sialic acid decreased fivefold the binding affinity. By competitive ELISA, peptides containing the N-linked oligosaccharide sequence inhibited the binding of the parent glycoprotein to immobilized AWN-1 5-45 times less effectively than those carrying O-linked NeuAc alpha(2-3/6)-Gal beta(1-3)-GalNAc structures. In addition, AWN-1 bound with a Kd = 0.3 microM to solubilized, biotinylated porcine zona pellucida glycoproteins. PDC-109 competed effectively with zona pellucida glycoproteins for AWN binding, whereas fetuin was a poor competitor. On the other hand, AWN epitopes were demonstrated on in vitro capacitated boar spermatozoa which were able to bind to, and penetrate, zona-encased oocytes. These data indicate that spermadhesin AWN-1 may play a role in pig fertilization as a sperm-associated lectin of broad specificity though preferential affinity for certain O-linked oligosaccharide structures of the oocyte's zona pellucida.
Dostálová
Z
Z
Institut für Reproduktionsmedizin, Tierärztliche Hochschule Hannover, Germany.
Calvete
J J
JJ
Sanz
L
L
Töpfer-Petersen
E
E
D000595:Amino Acid Sequence; D000818:Animals; D002240:Carbohydrate Sequence; D002352:Carrier Proteins / Q000378:metabolism*; D005260:Female; D008969:Molecular Sequence Data; D009844:Oligosaccharides / Q000378:metabolism*; D009865:Oocytes / Q000378:metabolism; D029589:Seminal Plasma Proteins; D013552:Swine; D015044:Zona Pellucida / Q000378:metabolism*
D002352:Carrier Proteins; D009844:Oligosaccharides; D029589:Seminal Plasma Proteins; C425042:spermadhesin