College of Chemistry and Chemical Engineering, State Key Laboratory of Applied Organic Chemistry, Lanzhou University, Lanzhou 730000, PR China.
Biochemistry. 2010 Nov 16;49(45):9839-47. doi: 10.1021/bi101305b. Epub 2010 Oct 26.
Glycosaminoglycans (GAGs) are known to participate in central nervous system processes such as development, cell migration, and neurite outgrowth. In this paper, we report an initial glycomics study of GAGs from the porcine central nervous system. GAGs of the porcine central nervous system, brain and spinal cord were isolated and purified by defatting, proteolysis, anion-exchange chromatography, and methanol precipitation. The isolated GAG content in brain was 5 times higher than in spinal cord (0.35 mg/g of dry sample, compared to 0.07 mg/g of dry sample). In both tissues, chondroitin sulfate (CS) and heparan sulfate (HS) were the major and the minor GAG, respectively. The average molecular masses of CS from brain and spinal cord were 35.5 and 47.1 kDa, respectively, and those for HS from brain and spinal cord were 56.9 and 34 kDa, respectively. The disaccharide analysis showed that the compositions of CS from brain and spinal cords are similar, with uronic acid (1→3) 4-O-sulfo-N-acetylgalactosamine residue corresponding to the major disaccharide unit (CS type A) along with five minor disaccharide units. The major disaccharides of both brain and spinal cord HS were uronic acid (1→4) N-acetylglucosamine and uronic acid (1→4) 6-O-sulfo-N-sulfoglucosamine, but their composition of minor disaccharides differed. Analysis by (1)H and two-dimensional NMR spectroscopy confirmed these disaccharide analyses and provided the glucuronic/iduronic acid ratio. Finally, both purified CS and HS were biotinylated and immobilized on BIAcore SA biochips. Interactions between these GAGs and fibroblast growth factors (FGF1 and FGF2) and sonic hedgehog (Shh) were investigated by surface plasmon resonance.
糖胺聚糖 (GAGs) 已知参与中枢神经系统的发育、细胞迁移和神经突生长等过程。在本文中，我们报告了猪中枢神经系统 GAG 的初步糖组学研究。通过脱脂、蛋白水解、阴离子交换色谱和甲醇沉淀从猪中枢神经系统、脑和脊髓中分离和纯化 GAG。脑 GAG 的分离含量是脊髓的 5 倍（0.35mg/g 干样品，相比之下，0.07mg/g 干样品）。在这两种组织中，硫酸软骨素 (CS) 和硫酸乙酰肝素 (HS) 分别是主要和次要的 GAG。脑和脊髓 CS 的平均分子量分别为 35.5 和 47.1 kDa，脑和脊髓 HS 的分子量分别为 56.9 和 34 kDa。二糖分析表明，脑和脊髓 CS 的组成相似，主要二糖单元（CS 型 A）对应的是带有五个次要二糖单元的 4-O-磺基-N-乙酰半乳糖胺残基。脑和脊髓 HS 的主要二糖均为葡萄糖醛酸（1→4）N-乙酰葡萄糖胺和葡萄糖醛酸（1→4）6-O-磺基-N-磺基葡萄糖胺，但它们的次要二糖组成不同。（1）H 和二维 NMR 光谱分析证实了这些二糖分析，并提供了葡萄糖醛酸/艾杜糖醛酸比。最后，纯化的 CS 和 HS 均被生物素化并固定在 BIAcore SA 生物芯片上。通过表面等离子体共振研究了这些 GAG 与成纤维细胞生长因子（FGF1 和 FGF2）和 sonic hedgehog（Shh）之间的相互作用。
