Structure and function of the zona pellucida: identification and characterization of the proteins of the mouse oocyte's zona pellucida.

摘要: Abstract The zona pellucida is an acellular coat which surrounds the plasma membrane of fully grown mammalian oocytes and performs a variety important functions during oogenesis, fertilization, preimplantation development. In this investigation proteins mouse oocyte's have been identified characterized by using zonae pellucidae isolated individually from with mouth-operated micropipets. Various morphological biochemical criteria were employed to assess purity and, in most cases, they found be virtually free contamination other oocyte proteins. It was determined that each contains 4.8 ng protein, represents 80% or more dry weight about 17% total protein. Electrophoretic analyses as few five treated diazotized [ 125 I]iodosulfanilic acid revealed presence only three radiolabeled proteins, designated ZP1, ZP2, ZP3. same Coomassie blue staining when large numbers (approximately 750) subjected SDS-polyacrylamide gel electrophoresis. These migrate broad bands on gels, consistent their being glycoproteins, apparent molecular weights 200,000 (ZP1), 120,000 (ZP2), 83,000 (ZP3). intact acid, reagent does not penetrate membrane, 3 H-labeled 1-dimethylaminonaphthalene-5-sulfonyl chloride. Results amino analysis high-resolution two-dimensional electrophoresis individual suggest protein unique polypeptide chain. ZP3 represent 36, 47, 17%, respectively, pellucida. reducing agents cause dissolution pellucida, ZP1 converted into species migrates 130,000, suggesting it exists oligomer, stabilized disulfide bonds, unreduced state. results these experiments are discussed terms properties before after fertilization compared obtained vitelline envelopes eggs nonmammalian animal species.