Changes in the specificity of the interaction between cell growth/differentiation factors and glycosaminoglycans during development and differentiation.

糖胺聚糖(GAG)是存在于动物组织中的一种具有重复二糖单元的骨架结构的复杂的硫酸盐多糖。在GAG链中发现的高度异质性的硫酸盐化模式以及GAG与蛋白质因子的特定分子相互作用参与了各种生物过程的调节。为了阐明发育过程中精细结构的变化与生长因子结合特异性的变化之间的关系，进行了以下实验。此外，为了更好地了解硫酸软骨素(CS)链的结构与功能关系，还建立了一种高灵敏度的测定低pmo1下CS寡糖序列的方法。从非洲爪哇4个不同发育阶段的胚胎中分离得到乙酰肝素硫酸酯和CS/皮肤素硫酸酯链，并比较了它们的结构和结合生长因子的能力。用BlAcore系统…进行生长因子结合实验更多的证据表明，GAG以类似的亲和力结合了一些生长因子，然而，在发育过程中，这些亲和力有很大的不同。这些结果表明，与生长因子特异相互作用的糖序列的表达可能在非洲爪哇胚胎发育过程中受到调控。并对其进行了验证和表征。抗GAG抗体免疫染色表明，抗软骨素抗体473Al2的表位定位于发育中的线虫包囊中，线虫包囊是线虫特有的刺细胞器。软骨素或CS蛋白多糖可能为线囊成分的组装提供了环境，也可能是线囊的成分。从鲨鱼软骨CS-C和鱿鱼软骨CS-E中分离出多种新的八糖和十糖，并用新开发的测序方法测定了它们的糖序列。这些结构定义的寡糖被用于表征几种广泛使用的抗CS抗体的表位。较少

Glycosaminoglycans (GAGs) are complex sulfated polysaccharides with a backbone structure composed of a repeating disaccharide unit and occur in animal tissues. Highly heterogeneous sulfated patterns found in the GAG chains and the specific molecular interactions of GAGs with protein factors are involved in the regulation of various biological processes. To elucidate the relation between alterations in the fine structure and the changes in the specificity of the growth factor-binding during development, the following experiments were carried out. Furthermore, to better understand the structure-function relation of chondroitin sulfate (CS) chains, the highly sensitive method to determine CS oligosaccharide sequences at low pmol was also developed.1. Heparan sulfate and CS/dermatan sulfate chains were isolated from four different stages of Xenopus laevis embryos and their structure and growth factor-binding capacities were compared. Growth factor-binding experiments using a BlAcore system … More demonstrated that GAGs bound some growth factors with comparable affinities, which are, however, significantly varied during development. These results suggested that expression of saccharide sequences, which specifically interact with a growth factor, might be regulated during Xenopus embryogenesis.2. GAGs of Hydra magnipapillata were demonstrated and characterized. Immunostaining using anti-GAG antibodies showed that the epitope of anti-chondroitin antibody 473Al2 was localized in the developing nematocysts, which is a sting organelle specific to cnidarians. The chondroitin or CS proteoglycans might furnish an environment for assembling nematocyst components, and might be components of nematocysts.3. A variety of novel octa-and decasaccharides were isolated from shark cartilage CS-C as well as squid cartilage CS-E, and their saccharide sequences were determined using the newly developed sequencing method. These structurally-defined oligosaccharides were applied for characterization of the epitopes of several widely used anti-CS antibodies. Less