Laboratory of Medical and Functional Glycomics, Graduate School of Advanced Life Science, Frontier Research Center for Post-Genome Science and Technology, Hokkaido University, Sapporo, Japan.
Anal Chem. 2011 Dec 15;83(24):9443-9. doi: 10.1021/ac2021079. Epub 2011 Nov 16.
Glycosaminoglycans (GAGs) play important roles in cell adhesion and growth, maintenance of extracellular matrix (ECM) integrity, and signal transduction. To fully understand the biological functions of GAGs, there is a growing need for sensitive, rapid, and quantitative analysis of GAGs. The present work describes a novel analytical technique that enables high throughput cellular/tissue glycosaminoglycomics for all three families of uronic acid-containing GAGs, hyaluronan (HA), chondroitin sulfate (CS)/dermatan sulfate (DS), and heparan sulfate (HS). A one-pot purification and labeling procedure for GAG Δ-disaccharides was established by chemo-selective ligation of disaccharides onto high density hydrazide beads (glycoblotting) and subsequent labeling by fluorescence. The 17 most common disaccharides (eight comprising HS, eight CS/DS, and one comprising HA) could be separated with a single chromatography for the first time by employing a zwitter-ionic type of hydrophilic-interaction chromatography column. These novel analytical techniques were able to precisely characterize the glycosaminoglycome in various cell types including embryonal carcinoma cells and ocular epithelial tissues (cornea, conjunctiva, and limbus).
糖胺聚糖（GAGs）在细胞黏附和生长、细胞外基质（ECM）完整性的维持以及信号转导中发挥着重要作用。为了充分了解 GAGs 的生物学功能，人们越来越需要对 GAGs 进行敏感、快速和定量的分析。本研究描述了一种新的分析技术，该技术可实现所有三种含糖醛酸的 GAG（透明质酸（HA）、硫酸软骨素/硫酸皮肤素（CS/DS）和硫酸乙酰肝素（HS））的高通量细胞/组织糖胺聚糖组学分析。通过糖选择性地将二糖连接到高密度酰肼珠上（糖印迹），并随后通过荧光标记，建立了 GAG Δ-二糖的一锅式纯化和标记程序。首次采用两性离子型亲水相互作用色谱柱，可通过单次色谱分离出 17 种最常见的二糖（8 种构成 HS，8 种 CS/DS，1 种构成 HA）。这些新的分析技术能够精确地描述包括胚胎癌细胞和眼部上皮组织（角膜、结膜和缘）在内的各种细胞类型中的糖胺聚糖组。
