Immunoglobulins against gp273, the ligand for sperm-egg interaction in the mollusc bivalve Unio elongatulus, are directed against charged O-linked oligosaccharide chains bearing a Lewis-like structure and interact with epitopes of the human zona pellucida.
In oocytes of the mollusc bivalve Unio elongatulus, gp273 is the ligand molecule for sperm-egg interaction and binding is mediated by its O-glycans. A serum raised against this protein enabled its localization in the crater region, the area of the vitelline coat where sperm recognition occurs, and showed that after cyanogen bromide fragmentation, the anti-gp273 epitope(s) was retained by a peptide where the O-glycans are localized. In this article, we utilized purified anti-gp273 immunoglobulins to characterize the corresponding epitope by: (i) immunoblotting analysis of the protein after removal of O- and N-glycans; (ii) solid phase binding analysis of anti-gp273 IgG to gp273 N- and O-glycans; and (iii) binding analysis of the same antibody to commercially available oligosaccharides. The results showed that the epitope consists of O-glycans and contains a Lewis-like structure with fucose as determinant. Anti-gp273 IgG were then used to investigate human zona pellucida by immunoelectronmicroscopy and immunoblotting. Epitopes recognized by the antibody were demonstrated on the outer surface of the zona pellucida and shown to belong to a zona pellucida protein having electrophoretic mobility similar to human ZP3. Since human sperm specifically bind to gp273, and anti-gp273 interferes with this binding a functional role for these epitopes is suggested.
Focarelli
Riccardo
R
Department of Evolutionary Biology, University of Siena, Siena, Italy. focarelli@unisi.it
Capone
Antonietta
A
Ermini
Leonardo
L
Del Buono
Francesca
F
Battista La Sala
Giovanni
G
Balasini
Marcella
M
Rosati
Floriana
F
D000818:Animals; D000906:Antibodies / Q000276:immunology*; D049872:Bivalvia / Q000378:metabolism*; D000939:Epitopes / Q000276:immunology; D005260:Female; D006023:Glycoproteins / Q000276:immunology*; D006801:Humans; D008297:Male; D009844:Oligosaccharides / Q000276:immunology*; D010063:Ovum / Q000378:metabolism; D013094:Spermatozoa / Q000378:metabolism; D015044:Zona Pellucida / Q000276:immunology
D000906:Antibodies; D000939:Epitopes; D006023:Glycoproteins; D009844:Oligosaccharides