Bakos M A, Kurosky A, Hedrick J L
Department of Biochemistry and Biophysics, University of California, Davis 95616.
Biochemistry. 1990 Jan 23;29(3):609-15. doi: 10.1021/bi00455a003.
Previous studies have shown that the Xenopus laevis egg envelope exists in three forms with differing ultrastructural, macromolecular, and sperm penetrability properties. The coelomic envelope (CE) is derived from eggs released from the ovary into the body cavity of the female, the vitelline envelope (VE) from eggs which have passed through the oviduct, and the fertilization envelope (FE) from fertilized eggs. In the present study, the physicochemical characteristics of these three envelope types were differentiated. Investigation of envelope solubility, deformability, sulfhydryl reactivity, and hydrophobic dye and ferritin binding capacity demonstrated that profound physicochemical changes occur in envelope conversions CE----VE----FE. The physical strength of the envelopes, as evidenced by deformability studies, ranked FE greater than CE greater than VE. These differences were not accountable by differences in the number of disulfide bonds, although the CE sulfhydryl groups were significantly less accessible than those in the VE or FE. All three envelope forms were hydrophilic in nature, exhibiting little ability to bind 1-anilino-8-naphthalenesulfonic acid. The CE bound greater amounts of ferritin in comparison to the VE and FE, indicating the presence of a basic domain, presumably in the 43-kDa glycoprotein, which is lost upon proteolysis to 41 kDa during the CE----VE conversion. The envelope integrity of all three forms was maintained by both noncovalent and covalent (disulfide) bonds. Measurements of the effect of pH on envelope solubilization indicated the involvement of an ionizable group with pKa of 8.0 in maintaining envelope structure.
先前的研究表明,非洲爪蟾的卵膜存在三种形式,它们具有不同的超微结构、大分子结构和精子穿透性。体腔膜(CE)源自从卵巢释放到雌性体腔中的卵,卵黄膜(VE)源自已经通过输卵管的卵,受精膜(FE)源自受精卵。在本研究中,对这三种膜类型的物理化学特性进行了区分。对膜的溶解性、可变形性、巯基反应性以及疏水染料和铁蛋白结合能力的研究表明,在膜的转化过程CE→VE→FE中发生了深刻的物理化学变化。通过可变形性研究证明,膜的物理强度排序为FE>CE>VE。尽管CE的巯基比VE或FE中的巯基明显更难接近,但这些差异不能用二硫键数量的差异来解释。所有三种膜形式本质上都是亲水的,几乎没有结合1-苯胺基-8-萘磺酸的能力。与VE和FE相比,CE结合了更多的铁蛋白,这表明存在一个碱性结构域,可能存在于43 kDa的糖蛋白中,在CE→VE转化过程中经蛋白水解为41 kDa时该结构域会丢失。所有三种形式的膜完整性都是通过非共价键和共价键(二硫键)维持的。pH对膜溶解作用的测量表明,一个pKa为8.0的可电离基团参与维持膜结构。