A role for the migrating sperm surface antigen PH-20 in guinea pig sperm binding to the egg zona peilucida

Abstract

After the acrosome reaction, the PH-20 surface antigen of guinea pig sperm migrates from its original location on the posterior head surface to a new location on the inner acrosomal membrane (Myles, D. G., and P. Primakoff, 1984, J. Cell Biol., 99: 1634-1641). We have isolated three monoclonal antibodies (MAbs) of the IgG1 subclass, PH-20, PH-21, and PH-22, that bind to the PH-20 antigen. The PH-20 MAb strongly inhibited (~90%) sperm binding to the guinea pig egg zona pellucida at saturating antibody concentrations (>20 µg/ ml). Half-maximal inhibition of sperm binding to the zona was obtained with ~2 µg/ml PH-20 MAb. The PH-21 MAb at saturating concentration (50 µg/ml) partially inhibited (~45%) spermzona binding, and the PH-22 MAb (50 µg/ml) did not inhibit (0%) sperm-zona binding. Essentially the same amounts of the three MAbs were bound to sperm under the conditions where inhibition (PH-20, PH-21) or no inhibition (PH-22) of sperm-zona binding was observed, which indicates that the different levels of inhibition did not arise from different levels of MAb binding. Competition binding assays with 125I-labeled MAbs showed that PH-21 binding to sperm was not affected by the binding of PH-20 or PH-22. However, that PH-20 and PH-22 blocked each other’s binding to sperm suggests that their recognized determinants may be relatively close to one another. The results indicate that the migrating PH-20 antigen has a required function in sperm binding to the zona pellucida and that the PH-20 MAb affects its active site. © 1985, Rockefeller University Press., All rights reserved.