Structural and Functional Analysis of glycoproteins participating in the mammalian fertilization and development

参与哺乳动物受精和发育的糖蛋白的结构和功能分析

• 批准号: 18570103
• 负责人: NAKANO Minoru
• 金额: $ 2.55万
• 依托单位: Chiba University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 2006
• 资助国家: 日本
• 起止时间: 2006 至 2007
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-18570103/
• 关键词: Zona Pellucida; Mammalian Ovum; Glycoprotein; Glycolipid; In vitro fertilization; Sperm-egg binding; Placenta; Pregnancy Associated Glycoprote; 免疫回避; 精子・卵子間結合

1) Binding of porcine and bovine sperm to glycolipid analogues We prepared plastic plates to which six kinds of glycolipid analogues having a monosaccharide at the end of alkyl chain are attached. To these plates, porcine or bovine sperm were added. After washing the plates, the numbers of residual sperm were counted. By this method, it was revealed that porcine sperm mainly bind to β-galactose residue of the glycolipid analogue, while bovine sperm to α-mannose residue. Furthermore, the acrosome-reacted sperm of both species bind to α-mannose residue.2) Binding of porcine and bovine sperm to the egg zona pellucida It is universally accepted that sperm species-specifically bind to the zona pellucida. Here we investigated whether porcine and bovine sperm bind to the zona pellucida of heterologous egg. As a result, it was shown that almost the same number of the sperm bound to the heterologous zona as that of the homologous sperm did, reflecting the distribution of the carbohydrate chains on the surface of the zona.3) The binding activity of recombinant zona pellucida glycoproteins to sperm Three zona glycoprotein components (ZPA, ZPB, ZPC) and their combinations were expressed in Sf9 cells and the binding activities of the recombinants to porcine and bovine sperm were analyzed. Since the high-mannose type chains are attached to the proteins by this expression system, the recombinants were bound not to porcine sperm but to bovine sperm. Any single component was not bound to bovine sperm and ZPB/ZPC coexpressed was bound.4) Determination of the carbohydrate structure of glycoproteins expressed at pregnancy The structure of main N-linked chain of bovine pregnancy associated glycoproteins was determined. The N-glycan has an unusual structure having GalNAc.

1）猪和牛精子与糖脂类似物的结合我们制备了塑料板，塑料板上六种类型的糖脂类似物在烷基链末端的末端具有单糖。在这些板上，加入了猪或牛精子。洗涤板后，计数残留精子的数量。通过这种方法，揭示了猪精子主要与糖脂类似物的β-半乳糖残基结合，而牛精子与α-甘露糖残基。此外，这两种物种的基础化体反应精子与α-甘露糖的退休结合。2）猪和牛精子与卵Zona pellucida的结合，普遍认为，精子物种特异性地结合了Zona Pellucida。在这里，我们研究了猪和牛精子是否与异源卵的Zona卵黄结合。结果，结果表明，几乎与同源精子绑定到异源Zona的精子数量相同，反映了Zona表面上的碳水化合物链的分布。分析了重组者与猪和牛精子的结合活性。由于该表达系统将高甘露糖型链连接到蛋白质上，因此重组剂与猪精子无关，而与牛精子结合。任何单个成分与牛精子无结合，ZPB/ZPC共表达均被结合。4）确定妊娠时表达的糖蛋白的碳氢化物结构。确定牛妊娠妊娠链的主要N-连接链的结构。 N-Glycan具有具有GALNAC的异常结构。

项目成果

Recombinant bovine zona pellucida glycoproteins ZP3 and ZP4 coexpressed in Sf9 cells form a snerm-binding active hetero-complex

在Sf9细胞中共表达的重组牛透明带糖蛋白ZP3和ZP4形成神经结合活性异源复合物

• 发表时间: 2007
• 期刊: FEBS Journal 274
• 作者: Saeko;Kanai
• 通讯作者: Kanai

Disulfide linkage patterns of pig zona pellucida glycoproteins ZP3 and ZP4

猪透明带糖蛋白ZP3和ZP4的二硫键连接模式

• 发表时间: 2008
• 期刊: Molecular Reproduction and Development 75
• 作者: Saeko Kanai;Tetsushi Kitayama;Naoto Yonezawa;Yoriko Sawano;Masaru Tanokura;Minoru Nakano
• 通讯作者: Minoru Nakano

Species-selective recognition of sperm by zona pellucida glycoproteins viacarbohydrate chains in pig and bovine

猪和牛的透明带糖蛋白通过碳水化合物链对精子进行物种选择性识别

• 发表时间: 2006
• 作者: Naoto;Yonezawa
• 通讯作者: Yonezawa

Recombinant bovine zona pellucida glycoproteins ZP3 and ZP4 coexpressed in Sf9 cells form a sperm-binding active hetero-complex

• DOI: 10.1111/j.1742-4658.2007.06065.x
• 发表时间: 2007-10-01
• 期刊: FEBS JOURNAL
• 影响因子: 5.4
• 作者: Kanai, Saeko;Yonezawa, Naoto;Nakano, Minoru
• 通讯作者: Nakano, Minoru

Society of Reproduction and Fertility Supplement63

生殖和生育补充协会63

• 发表时间: 2006
• 作者: Naoto;Yonezawa
• 通讯作者: Yonezawa