Laboratory of Proteoglycan Signaling and Therapeutics, Graduate School of Life Science, Hokkaido University, Frontier Research Center for Post-Genomic Science and Technology, Nishi 11-choume, Kita 21-jo, Kita-ku, Sapporo, Hokkaido, 001-0021, Japan.
Glycoconj J. 2010 Jan;27(1):49-60. doi: 10.1007/s10719-009-9253-x. Epub 2009 Aug 29.
The amniotic membrane (AM) is the innermost layer of fetal membranes and possesses various biological activities. Although the mechanism underlying these biological activities remains unclear, unique components seem to be involved. AM contains various extracellular matrix components such as type I collagen, laminin, fibronectin, hyaluronan, and proteoglycans bearing chondroitin sulfate/dermatan sulfate (CS/DS) glycosaminoglycan side chains. Since CS/DS have been implicated in various biological processes, we hypothesized that CS/DS in AM may play a major role in the biological activities of AM. Therefore, the structure and bioactivity of the CS/DS chains from porcine fetal membranes (FM-CS/DS) were investigated. A compositional analysis using various chondroitinases revealed that the characteristic DS domain comprised of iduronic acid-containing disaccharide units is embedded in FM-CS/DS, along with predominant disaccharide units, GlcA-GalNAc, GlcA-GalNAc(4-O-sulfate), and GlcA-GalNAc(6-O-sulfate), where GlcA and GalNAc represent D-glucuronic acid and N-acetyl-D-galactosamine, respectively. The average molecular mass of FM-CS/DS chains was unusually large and estimated to be 250 - 300 kDa. The FM-CS/DS chains showed neurite outgrowth-promoting activity, which was eliminated by digestion with chondroitinase ABC of the CS/DS chains. This activity was suppressed by antibodies against growth factors including pleiotrophin, midkine, and fibroblast growth factor-2, suggesting the involvement of these growth factors in the neurite outgrowth-promoting activity. The binding of these growth factors to FM-CS/DS was also demonstrated by surface plasmon resonance spectroscopy.
羊膜（AM）是胎儿膜的最内层，具有各种生物活性。尽管这些生物活性的机制尚不清楚，但似乎涉及到独特的成分。AM 含有各种细胞外基质成分，如 I 型胶原、层粘连蛋白、纤维连接蛋白、透明质酸和带有软骨素硫酸盐/硫酸皮肤素（CS/DS）糖胺聚糖侧链的蛋白聚糖。由于 CS/DS 参与了各种生物过程，我们假设 AM 中的 CS/DS 可能在 AM 的生物活性中起主要作用。因此，研究了来自猪胎儿膜（FM-CS/DS）的 CS/DS 链的结构和生物活性。使用各种软骨素酶的组成分析表明，特征性的 DS 结构域由含有艾杜糖醛酸的二糖单元组成，嵌入 FM-CS/DS 中，还存在主要的二糖单元，GlcA-GalNAc、GlcA-GalNAc(4-O-硫酸盐）和 GlcA-GalNAc(6-O-硫酸盐），其中 GlcA 和 GalNAc 分别代表 D-葡萄糖醛酸和 N-乙酰-D-半乳糖胺。FM-CS/DS 链的平均分子量非常大，估计为 250-300 kDa。FM-CS/DS 链具有促进神经突生长的活性，这种活性可以通过 CS/DS 链的软骨素酶 ABC 消化消除。这种活性被针对包括多效蛋白、中期因子和碱性成纤维细胞生长因子-2 在内的生长因子的抗体抑制，表明这些生长因子参与了促进神经突生长的活性。表面等离子体共振光谱也证明了这些生长因子与 FM-CS/DS 的结合。
