Regulation of extracellular matrix assembly: in vitro reconstitution of a partial fertilization envelope from isolated components.
At fertilization, the glycocalyx (vitelline layer) of the sea urchin egg is transformed into an elevated fertilization envelope by the association of secreted peptides and the formation of intermolecular dityrosine bonds. Dityrosine cross-links are formed by a secreted ovoperoxidase that exists in a Ca2+-stabilized complex with proteoliaisin in the fertilization envelope. By using purified proteins, we now show that proteoliaisin is necessary and sufficient to link ovoperoxidase to the egg glycocalyx. Specifically, we have found that ovoperoxidase can associate with the vitelline layer only when complexed with proteoliaisin; proteoliaisin binds to the vitelline layer independently of its association with ovoperoxidase; proteolytic modification of the vitelline layer is not required for this interaction to occur; the binding of proteoliaisin to the vitelline layer is mediated by the synergistic action of the two major seawater divalent cations, Ca2+ and Mg2+; the number of proteoliaisin-binding sites on the vitelline layer of unfertilized eggs is equivalent to the amount of proteoliaisin secreted at fertilization; and the binding of ovoperoxidase to the vitelline layer, via proteoliaisin, permits the in vitro cross-linking of these two in vivo substrates. The association of purified ovoperoxidase and proteoliaisin with the vitelline layer of unfertilized eggs reconstitutes part of the morphogenesis of the fertilization envelope.
D000818:Animals; D002118:Calcium / Q000494:pharmacology; D005109:Extracellular Matrix / Q000378:metabolism*; D005260:Female; D005306:Fertilization; D006023:Glycoproteins; D008274:Magnesium / Q000494:pharmacology; D008562:Membrane Glycoproteins; D008565:Membrane Proteins / Q000502:physiology; D009024:Morphogenesis; D010063:Ovum / Q000378:metabolism* / Q000648:ultrastructure; D010544:Peroxidases / Q000502:physiology; D011134:Polysaccharides; D011485:Protein Binding / Q000187:drug effects; D012617:Sea Urchins / Q000196:embryology; D014443:Tyrosine / Q000378:metabolism; D014817:Vitelline Membrane / Q000378:metabolism*
D006023:Glycoproteins; D008562:Membrane Glycoproteins; D008565:Membrane Proteins; D011134:Polysaccharides; C044807:proteoliaisin protein, Strongylocentrotus purpuratus; D014443:Tyrosine; D010544:Peroxidases; D008274:Magnesium; D002118:Calcium