Intermolecular cross-linking of vitelline envelope polypeptides predominates in the hardened sea urchin fertilization envelope.
At fertilization, the sea urchin egg vitelline envelope (VE) elevates, and a subset of released cortical granule proteins, paracrystalline protein fraction (PCF), associates with the VE to form the fertilization envelope (FE). Cortical granule peroxidase cross-links FE polypeptides by phenolic coupling of tyrosyl residues. We have used an immunological approach to determine which polypeptides are linked together in the hardened FE of Strongylocentrotus purpuratus. Soluble polypeptides were extracted from hardened FEs, and antibodies were prepared in rabbits against the insoluble envelope matrix (FE ghost). Whole immune serum and purified IgGs each reacted with FE ghosts when using an enzyme-linked immunosorbent assay. VEs isolated by means of three published procedures cross-reacted with the immune serum and purified IgGs. Soluble FE polypeptides also cross-reacted with whole immune serum and IgGs owing to the presence of VE polypeptides. Hyalin, a protein not found in FEs, and PCF did not cross-react with antiserum against FE ghosts. To determine which VE polypeptides were cross-linked in the hardened FE, VE polypeptides were immunoblotted by using antiserum against FE ghosts. Most of the VE polypeptides that ranged from 68,000 to 283,000 molecular weight cross-reacted with the antibody.
D000818:Animals; D000906:Antibodies; D000936:Antigen-Antibody Complex / Q000032:analysis; D003429:Cross Reactions; D003432:Cross-Linking Reagents; D004797:Enzyme-Linked Immunosorbent Assay; D005260:Female; D005306:Fertilization; D007074:Immunoglobulin G / Q000302:isolation & purification; D008297:Male; D008565:Membrane Proteins / Q000032:analysis* / Q000276:immunology; D010455:Peptides / Q000032:analysis* / Q000276:immunology; D012617:Sea Urchins / Q000502:physiology*; D014817:Vitelline Membrane / Q000032:analysis*
D000906:Antibodies; D000936:Antigen-Antibody Complex; D003432:Cross-Linking Reagents; D007074:Immunoglobulin G; D008565:Membrane Proteins; D010455:Peptides