Department of Physiology, School of Medicine, Toho University, 5-21-16 Ohmori-nishi, Ohta-ku, Tokyo 143-8540.
Department of Medical Education, School of Medicine, Kitasato University, 1-15-1 Kitasato, Sagamihara, Kanagawa 228-8555.
J Biol Chem. 2010 May 14;285(20):15627-15636. doi: 10.1074/jbc.M109.079483. Epub 2010 Mar 18.
Fertilization comprises oligosaccharide-mediated sperm-egg interactions, including sperm binding to an extracellular egg envelope, sperm penetration through the envelope, and fusion with an egg plasma membrane. We show that Xenopus dicalcin, an S100-like Ca(2+)-binding protein, present in the extracellular egg envelope (vitelline envelope (VE)), is a suppressive mediator of sperm-egg interaction. Preincubation with specific antibody greatly increased the efficiency of in vitro fertilization, whereas prior application of exogenous dicalcin substantially inhibited fertilization as well as sperm binding to an egg and in vitro sperm penetration through the VE protein layer. Dicalcin showed binding to protein cores of gp41 and gp37, constituents of VE, in a Ca(2+)-dependent manner and increased in vivo reactivity of VE with a lectin, Ricinus communis agglutinin I, which was accounted for by increased binding ability of gp41 to the lectin and greater exposure of gp41 to an external environment. Our findings strongly suggest that dicalcin regulates the distribution of oligosaccharides within the VE through its binding to the protein core of gp41, probably by modulating configuration of oligosaccharides on gp41 and the three-dimensional structure of VE framework, and thereby plays a pivotal role in sperm-egg interactions during fertilization.
受精包括糖基化介导的精子-卵子相互作用,包括精子与细胞外卵包膜的结合、精子穿透包膜以及与卵质膜的融合。我们发现,Xenopus dicalcin(一种存在于细胞外卵包膜(卵黄包膜(VE)中的 S100 样 Ca(2+)结合蛋白)是精子-卵子相互作用的抑制性介质。与特异性抗体预孵育大大提高了体外受精的效率,而外源性 dicalcin 的预先应用则显著抑制了受精以及精子与卵子的结合和体外精子穿透 VE 蛋白层。Dicalcin 以 Ca(2+)依赖性方式与 VE 的组成部分 gp41 和 gp37 的蛋白核心结合,并增加了 VE 与凝集素蓖麻凝集素 I 的体内反应性,这归因于 gp41 与凝集素的结合能力增加以及 gp41 向外部环境的更大暴露。我们的研究结果强烈表明,dicalcin 通过与 gp41 的蛋白核心结合,调节 VE 内寡糖的分布,可能通过调节 gp41 上寡糖的构象和 VE 框架的三维结构,从而在受精过程中的精子-卵子相互作用中发挥关键作用。