Department of Laboratory Medicine, Institute of Biomedicine, Sahlgrenska Academy at the University of Gothenburg, Gothenburg, Sweden.
Department of Experimental Medical Science, Division of Neuroscience, Glycobiology Group, Lund University, Lund, Sweden.
Methods Mol Biol. 2022;2303:173-182. doi: 10.1007/978-1-0716-1398-6_15.
Although glycosaminoglycans (GAGs) are known to be involved in a variety of physiological and pathological processes, knowledge about their expression by cells or tissues, the GAGome, is limited. Xylosides can be used to induce the formation of GAGs without the presence of a proteoglycan core protein. The administration of xylosides to living cells tends to result in a considerable amplification in GAG production, and the xylosides can, therefore, be used as analytical tools to study the GAG produced by a certain cell type. One of the most common ways to analyze the GAGs structurally is by disaccharide analysis, which involves depolymerization of the GAGs into disaccharides, fluorescent labeling of the disaccharides with 2-aminoacridone, and quantification using high-pressure liquid chromatography (HPLC). Here, we describe the procedure of producing xyloside-primed GAGs and how to study them structurally by disaccharide analysis.
尽管糖胺聚糖 (GAGs) 已知参与多种生理和病理过程，但对细胞或组织中 GAG 的表达（即 GAGome）的了解有限。木糖苷可用于在没有核心蛋白聚糖的情况下诱导 GAG 的形成。木糖苷给药于活细胞往往会导致 GAG 产量的显著增加，因此，木糖苷可用作分析工具来研究特定细胞类型产生的 GAG。分析 GAG 结构的最常见方法之一是二糖分析，它涉及将 GAG 解聚成二糖，用 2-氨基吖啶酮对二糖进行荧光标记，然后使用高压液相色谱 (HPLC) 进行定量。在这里，我们描述了产生木糖苷引发的 GAG 的过程，以及如何通过二糖分析来研究它们的结构。
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