title: Studies on mouse sperm binding molecule (ligand) to egg zona pellucida. II. Purification of ligand molecule from cauda epididymal sperm.
abstract: One of the ligand molecules was purified from mouse cauda epididymal sperm for an indication of its inhibitory activity in sperm-egg binding. It was isolated in an electrophoretically homogeneous form by ionophore A23187 treatment of sperm, sedimentation by ultracentrifugation, solubilization by 2% Nonidet P-40 (NP-40), repeated ion exchange chromatography on DEAE-Sepharose CL-6B and gel filtration on Sephacryl S-200. The purified ligand exhibited a molecular mass of 60,000 dalton by gel filtration and 67,000 dalton by sodium dodecyl sulfate polyacrylamide-gel electrophoresis (SDS-PAGE). This molecule was radioiodinated by the lactoperoxidase method and assayed the binding ability to egg zona pellucida. The labeled ligand specifically bound to both cumulus-free egg zona pellucida and isolated-zona pellucida of unfertilized egg in a similar extent, but didn't bind to 2-cell egg zona pellucida. The maximal binding site of ligand per egg zona pellucida was estimated about 660000 molecules by Scatchard plot analysis.
authors.affiliation: Faculty of Pharmaceutical Sciences, Kobe-Gakuin University, Japan.
authors.forename: Y
authors.initials: Y
authors.lastname: Kawai
authors.forename: K
authors.initials: K
authors.lastname: Funakoshi
authors.forename: T
authors.initials: T
authors.lastname: Hama
authors.forename: T
authors.initials: T
authors.lastname: Mayumi
mesh_terms: D000818:Animals; D000001:Calcimycin / Q000494:pharmacology; D002848:Chromatography, DEAE-Cellulose; D002852:Chromatography, Ion Exchange; D004591:Electrophoresis, Polyacrylamide Gel; D004822:Epididymis / Q000737:chemistry / Q000166:cytology*; D005260:Female; D066298:In Vitro Techniques; D007700:Kinetics; D008024:Ligands; D008297:Male; D051379:Mice; D008813:Mice, Inbred ICR; D008970:Molecular Weight; D013084:Sperm-Ovum Interactions / Q000502:physiology*; D013094:Spermatozoa / Q000737:chemistry*; D015044:Zona Pellucida / Q000378:metabolism*; D001619:beta-N-Acetylhexosaminidases / Q000032:analysis
chemical_list: D008024:Ligands; D000001:Calcimycin; D001619:beta-N-Acetylhexosaminidases