Qualitative characterization of oligosaccharide chains present on the rat zona pellucida glycoconjugates.
Qualitative characterization of oligosaccharide chains present on the rat zona pellucida glycoconjugates.
复制标题
大鼠透明带糖缀合物上存在的寡糖链的定性表征。
DOI：
10.1095/biolreprod46.5.912
复制
发表时间：
1992
影响因子：
3.6
通讯作者：
Tulsiani,DR
中科院分区：
文献类型：
--
作者：
Araki,Y;Orgebin-Crist,MC;Tulsiani,DR
透明带(ZP)是环绕在哺乳动物卵母细胞周围的细胞外糖萼，被认为是介导物种特异性精卵相互作用的物质。尽管有许多关于ZP糖结合物在几个物种中的特征的研究，但关于大鼠ZP的各种成分的数量和化学性质的信息很少或根本没有。在这项研究中，我们尝试使用内糖水解酶和/或外糖水解酶对大鼠ZP进行生化表征。用氯胺-T法对超排大鼠的完整卵进行放射性碘标记，并在非还原条件下将标记的ZP组分在SDS-PAGE上进行分离。这些研究表明，大鼠ZP由三个组分组成，表观分子质量分别为205 kDa(ZP1)、119 kDa(ZP2)和115 kDa(ZP3)。与小鼠ZP2和ZP3不同，小鼠ZP2和ZP3在SDS-PAGE上分离为不同的组分，而大鼠ZP2和ZP3在分子大小和等电点上显示出很大的重叠。用外切酶(神经氨酸酶和α-L岩藻糖苷酶)和/或内切糖苷酶(内切糖苷酶H，内切糖苷酶F，N-糖苷酶，N-葡聚糖酶)处理大鼠ZP组分，结果表明：1)大鼠ZP_2和ZP_3都含有N-连接的寡糖(OS)单位，这表明它们对内切糖苷酶F和N-糖苷酶的敏感性。2)经N-葡聚糖酶处理后，大鼠ZP2和ZP3组分的大小分别减小了近50%和60%，表明这两种ZP组分是高度糖基化的。3)大鼠ZP3是敏感的To-葡聚糖酶，提示该ZP组分含有O-连接的OS单位(S)。4)大鼠ZP_3的O-连接OS单位(S)上未发现岩藻糖基或唾液酸基残基(S)，提示该ZP组分含有O-连接OS单位(S)。4)大鼠ZP3Theo连接的OS单元(S)上存在岩藻糖基或唾液酰基残基，未得到cvldc：ncc。讨论了哺乳动物ZP上OS链在精子-卵子识别和结合中的潜在作用。
Zona pellucida (ZP), the extracellular glycocalyx surrounding the mammalian oocyte, is believed to mediate species-specific sperm-egg interaction. Despite numerous studies on characterization of ZP glycoconjugates in several species, little or no information is available on the number and chemical nature of the various components of the rat ZP. In this study we have attempted the biochemical characterization of the rat ZP using endo- and/or exo-glycohydrolases. Intact eggs from superovulated rats were radioiodinated by the chloramine-T method, and the labeled ZP components were resolved on SDS-PAGE under nonreducing conditions. These studies show that the rat ZP consists of three components with apparent molecular masses of 205 kDa (ZP1), 119 kDa (ZP2), and 115 kDa (ZP3). Unlike mouse ZP2 and ZP3, which resolve as distinct components on SDS-PAGE, rat ZP2 and ZP3 show substantial overlap in their molecular sizes and isoelectric points. Treatment of the rat ZP components with exo- (neuraminidase and α-L-fucosidase) and/or endo- (endoglycosidase H, endoglycosidase F,N-glycanase, andO-glycanase) glycohydrolases indicated the following: 1) Both rat ZP2 and ZP3 contain N-linked oligosaccharide (OS) units as indicated by their sensitivity to endoglycosidase F andN-glycanase. 2) Treatment withN-glycanase caused a reduction in size of the rat ZP2 and ZP3 components by nearly 50% and 60%, respectively, indicating that the two ZP components are highly glycosylated. 3) Rat ZP3 was sensitive toO-glycanase, suggesting that this ZP component containsO-linked OS unit(s). 4) No evidence was obtained for the presence of fucosyl or sialyl residue(s) on theO-linked OS unit(s) of rat ZP3 was sensitive toO-glycanasc, suggesting that this ZP component containsO-linked OS unit(s). 4) No cvldc:ncc was obtained for the presence of fucosyl or sialyl residue{s) on theO-linked OS unit(s) of rat ZP3. The potential role of the OS chains present on mammalian ZP in sperm-egg recognition and binding is discussed.