Fourier Transform Infrared Spectroscopic Analysis of the Intact Zona Pellucida of the Mammalian Egg: Changes in the Secondary Structure of Bovine Zona Pellucida Proteins During Fertilization
Fourier Transform Infrared Spectroscopic Analysis of the Intact Zona Pellucida of the Mammalian Egg: Changes in the Secondary Structure of Bovine Zona Pellucida Proteins During Fertilization
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DOI：
10.1177/153537020623100206
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发表时间：
2006-02
影响因子：
3.2
通讯作者：
M. Nara;N. Yonezawa;Takeshi Shimada;Kazuya Takahashi;M. Tanokura;F. Yumoto;H. Nakagawa;K. Ohashi;S. Hamano;M. Nakano
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文献类型：
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作者：
M. Nara;N. Yonezawa;Takeshi Shimada;Kazuya Takahashi;M. Tanokura;F. Yumoto;H. Nakagawa;K. Ohashi;S. Hamano;M. Nakano
卵透明膜是哺乳动物卵母细胞外的透明膜。分析卵透明质酸蛋白质的结构变化对于理解卵透明质酸在受精和着床前的重要生理作用的分子机制是必不可少的。受精过程中精子结构的变化导致精子的硬化被普遍认为是阻碍多精受精的原因。在这项研究中，我们分析了在受精过程中的二级结构的傅里叶变换红外光谱（FTIR）和透射电子显微镜的变化。利用FTIR光谱确定了猪卵清蛋白在水中以β折叠结构为主。还存在α-受阻胺结构。完整的、不溶解的猪卵透明带的衰减全反射（ATR）-FTIR光谱表明，天然透明带中的蛋白质也以β-结构为主。用衰减全反射-傅里叶变换红外光谱法（ATR-FTIR）对牛卵巢卵和受精卵的透明细胞进行了研究，结果表明，在受精过程中，透明细胞的β-结构含量增加。此外，在受精过程中的厚度的减少，观察到使用透射电子显微镜。因此，在受精过程中导致精子硬化的精子结构的变化伴随着精子蛋白二级结构的变化。
The zona pellucida is the acellular transparent envelope surrounding the mammalian oocyte. An analysis of the changes in the structures of zona pellucida proteins is essential for understanding the molecular mechanisms underlying the important physiological roles of the zona during fertilization and prelmplantatlon. The hardening of the zona caused by the structural changes during fertilization is generally accepted to be responsible for blocking polyspermy. In this study, we analyzed changes in the secondary structure of the zona during fertilization by Fourier transform infrared (FTIR) spectroscopy and transmission electron microscopy. The predominance of β-sheet structure in porcine ovarian egg zona proteins in water was ascertained using FTIR spectra. α-Helix structure was also present. The attenuated total reflection (ATR)-FTIR spectrum of intact, unsolubilized porcine zonae pellucidae from ovarian eggs Indicated that the zona proteins in the native zona pellucida also have β-structure as the main constituent. Attenuated total reflectlon-FTIR spectroscopy of intact bovine zona pellucida obtained from ovarian and fertilized eggs at the blastocyst stage revealed that the β-structure content Increased during fertilization. Furthermore, a reduction of the thickness of the zona during fertilization was observed using transmission electron microscopy. Therefore, the change in the zona architecture that causes hardening of the zona during fertilization is accompanied by changes in the secondary structure of the zona proteins.