猪精子中与卵透明带糖蛋白ZP3结合的蛋白质

依次经ＰＳＬ－Ｓｅｐｈａｒｏｓｅ亲和层析柱和纤维素ＣＭ－５２离子交换层析柱，从猪精子的ＣＨＡＰＳ抽提液分离得４个蛋白质组分。用固相透明带精蛋白结合试验（ＩＺＰＧＢＡ）检测；表明精子蛋白ＳＰ１和ＳＰ２具有结合透明带糖蛋白ＺＰ３的活性，ＳＰ２并显示凝集血球的活性。精子蛋白ＳＰ１与卵预温育明显抑制精卵结合，抑制活性与加入的精子蛋白的浓度呈正相关。用生物素标记的ＺＰ３和蛋白质印迹技术，证明ＳＰ１中的６８ｋＤ精子蛋白与ＺＰ３结合，提示６８ｋＤ精子蛋白参与精卵结合。

The Pellucida Glycoprotein ZP3-Binding Protein The Pellucida Glycoprotein ZP3-Binding Protein of the Boar Spermatozoa

Four protein components have been isolated from the CHAPS extract of boar spermatozoa by chromatography on PSL-Sepharose affinity column and cellulose CM-52 ion exchange column sequentially. Both boar sperm protein SP1 and SP2 exhibited binding activity with the zona pellucida glycoprotein ZP3 by immobilized zona pellucida glycoprotein binding assay (IZPGBA), and SP2 also showed hemagglutinating activity. The protein SP1 obviously inhabited sperm-oocyte binding when it was preincubated with oocytes. The inhibitory activity of SP1 was related to its concentration. By using Western blot technique and biotinylated ZP3,a ZP3-binding spermatozoa protein with molecular weight of 68kD was identified, which may be a protein involved in sperm-oocyte binding.