Dev Biol 1997 Jul 15;1872:200-8. doi: 10.1006/dbio.1997.8608.

Mapping sperm binding domains on the sea urchin egg receptor for sperm.

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The receptor on the surface of the egg of the sea urchin Strongylocentrotus purpuratus that mediates species-specific binding of sperm is a 350-kDa cell surface glycoprotein. Earlier studies established that a recombinant protein encompassing a major portion of the N-terminal half of the receptor inhibited fertilization when tested in a competitive fertilization bioassay. To identify in more detail the sites in this domain of the receptor that are involved in binding sperm, a series of deletion constructs were expressed as glutathione S-transferase fusion proteins and tested for inhibitory activity in a fertilization bioassay. In addition, a novel assay for directly testing the sperm binding activity of these proteins was developed. In this assay we quantitated sperm binding to recombinant proteins representing various domains of the receptor immobilized on glutathione agarose beads. Using this new assay, two domains in the N-terminal half of the receptor were found to be involved in sperm binding. One of the peptide domains, composed of 247 amino acids, binds both the sperm of S. purpuratus and the sperm of another genus of sea urchin, Lytechinus pictus. In contrast, binding to the second domain consisting of a 32-amino-acid residue peptide was found to be genus specific; no binding of L. pictus sperm was observed. A working model is proposed incorporating these findings with earlier studies on the function of the oligosaccharide chains of the receptor. In this model it is postulated that the sperm initially interact with the nonspecific binding domain on the polypeptide and the sulfated O-linked oligosaccharide chains of the receptor. This interaction is followed by binding to the more specific polypeptide binding site on the receptor. It is proposed that only subsequent to binding at this second site can gamete fusion occur.

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Genes referenced: LOC100887844 LOC115919910 LOC752853