Isolation and biochemical characterization of a zona pellucida‐binding glycoprotein of boar spermatozoa

Abstract

Lectin‐like molecules on the sperm surface are implicated in the process of gamete recognition and adhesion. We have isolated and biochemically characterized a 15 kDa glycoprotein from ejaculated boar sperm which possess zona pellucida‐binding‐ and haemagglutinating‐activity. The zona/ 15 kDa protein interaction is inhibited by fucoldan, suggesting that the glycoprotein is one of the sperm components which participate in the initial gamete interaction. N‐Terminal sequence analysis of the isolated 15 kDa glycoprotein showed that it may belong to the same sperm/egg recognition‐mediating protein family as the sea urchin sperm protein binding.

Localization and structural characterization of an oligosaccharide O‐linked to bovine PDC‐109 Quantitation of the glycoprotein in seminal plasma and on the surface of ejaculated and capacitated spermatozoa

“…PDC-109 was isolated from either seminal plasma or acid extracts of ejaculated sperm [15] by reverse-phase HPLC using a SuperPac Pep-S (Pharmacia) eluting at 1 ml/min with a gradient of 0.1% (v/v) trifluoroacetic acid in (A) water and (B) acetonitrile. Chromatographic conditions were: isocratically (25% B) for 5 min, followed by 25-30% B for 5 min, 3@45% B for 60 min, and 45-70% B for 20 min.…”

“…The ability of purified PDC-109 to bind biotinylated ligands (zona pellucida glycoproteins, heparin, or soybean trypsin inhibitor) was assessed as in [15,18,19].…”

Electrophoretic and zymographic characterization of proteins isolated by various extraction methods from ejaculated and capacitated boar sperms

“…Isolation using OBG as a mild detergent most probably yielded membrane proteins, both peripheral and integral [25], but it is doubtful Electrophoresis 2011Electrophoresis , 32, 1309Electrophoresis -1318 whether intraacrosomal proteins were present as in the case when acidic extraction is performed (Fig. 1A; lane 3) and sperm surface proteins bound only by weak non-covalent interactions and intraacrosomal proteins are preferentially extracted [22,24]. SDS extracts (Fig.…”

“…This method is therefore excellent for acquisition of the acrosome content, but not adequate for the isolation of the membrane proteins only. This method is utilized for the isolation of surface proteins with ZP-binding activity [24].…”

Cloning of boar SPMI gene which is expressed specifically in seminal vesicle and codes for a sperm motility inhibitor protein

“…The spermadhesin family presently includes 11 members: PSP-1 (or SP3/SP6), PSP-2 (or SP4/SP7) [11], AWN, AQN-I, AQN-2, AQN-3 [12,13,[16][17]20], ACR-3 antigen (or Eja) [18], EJc, EJd, EJe and EJf [19]. The present data clearly indicate that, because of its 97% similarity to AQN-3 peptide, boar SPMI should became the 12th member reported of this family (Table 2).…”