SPECIFICITY OF BINDING OF ZONA-PELLUCIDA GLYCOPROTEINS TO SPERM PROACROSIN AND RELATED PROTEINS

A major regulatory site for species specificity of fertilization in mammals lies at the level of sperm binding to the zona pellucida. This implies a high degree of complementarity between gamete receptor molecules. One putative receptor molecule on spermatozoa is proacrosin/acrosin which binds to zona pellucida glycoproteins (ZPGPs) non-enzymically and with high affinity. The mechanism of recognition involves polysulphate groups in a particular stereochemical orientation and it has been suggested that this may contribute to species specificity of sperm-egg binding. In the present work this hypothesis has been tested by challenging I-125-labelled ZPGPs from pig, cow, and hamster eggs to recognize heterologous sperm proacrosins as well as a variety of sequence-related and unrelated proteins. Results show that pig and cow I-125-ZPGPs bind readily to boar, ram, and bull proacrosin but do not recognize guinea-pig proacrosin or any of the polysulphate binding proteins from rat, hamster, or mouse spermatozoa. Hamster I-125-ZPGPs also recognise boar, ram, and bull proacrosin as well as a range of unidentified proteins in pH3 extracts of hamster, rat, and mouse spermatozoa. The binding of ZPGPs to a variety of proteins not related to proacrosin is strongest to those with a high content of basic residues (i.e., pI > 8.5), although the secondary folding of the target protein is of major importance as boar proacrosin (pI 6.75) has the highest binding capacity of all proteins tested. Cross-reaction of ZPGP probes was not observed to guinea-pig proacrosin, suggesting that in this species the conformation of the protein is different to other sperm proacrosins. These results suggest that there are significant differences in the affinity of ZPGP-proacrosin and ZPGP-protein interactions to contribute to species specificity of sperm-egg binding.