Department of Chemistry and Chemical Biology, Center for Biotechnology and Interdisciplinary Studies, Rensselaer Polytechnic Institute, Troy, NY 12180, USA.
J Chromatogr A. 2012 Feb 17;1225:91-8. doi: 10.1016/j.chroma.2011.12.063. Epub 2011 Dec 26.
Glycosaminoglycans are a family of polysaccharides widely distributed in all eukaryotic cells. These polyanionic, linear chain polysaccharides are composed of repeating disaccharide units that are often differentially substituted with sulfo groups. The diversity of glycosaminoglycan structures in cells, tissues and among different organisms reflect their functional an evolutionary importance. Glycosaminoglycan composition and structure also changes in development, aging and in disease progression, making their accurate and reliable analysis a critical, albeit, challenging endeavor. Quantitative disaccharide compositional analysis is one of the primary ways to characterize glycosaminoglycan composition and structure and has a direct relationship with glycosaminoglycan biological functions. In this study, glycosaminoglycan disaccharides, prepared from heparan sulfate/heparin, chondroitin sulfate/dermatan sulfate and neutral hyaluronic acid using multiple polysaccharide lyases, were fluorescently labeled with 2-aminoacridone, fractionated into 17 well-resolved components by reverse-phase ultra-performance liquid chromatography, and analyzed by electrospray ionization mass spectrometry. This analysis was successfully applied to cell, tissue, and biological fluid samples for the picomole level detection of glycosaminoglycan composition and structure.
糖胺聚糖是广泛分布于所有真核细胞中的多糖家族。这些带负电荷的线性链多糖由重复的二糖单元组成，这些二糖单元通常带有不同数量的磺酸基团。细胞、组织和不同生物体中糖胺聚糖结构的多样性反映了它们的功能和进化重要性。糖胺聚糖的组成和结构在发育、衰老和疾病进展中也会发生变化，因此准确可靠地分析它们是一项至关重要但具有挑战性的任务。定量二糖组成分析是表征糖胺聚糖组成和结构的主要方法之一，与糖胺聚糖的生物学功能直接相关。在本研究中，使用多种多糖裂解酶从硫酸乙酰肝素/肝素、硫酸软骨素/硫酸皮肤素和中性透明质酸中制备糖胺聚糖二糖，并用 2-氨基吖啶酮进行荧光标记，通过反相超高效液相色谱分离成 17 个分辨率良好的组分，并通过电喷雾电离质谱进行分析。该分析方法成功应用于细胞、组织和生物体液样品，可用于检测糖胺聚糖组成和结构的皮摩尔水平。
