The Laboratory of Toxicology, Faculty of Pharmacy, Osaka Ohtani University, Osaka, Japan.
Methods Mol Biol. 2024;2763:159-169. doi: 10.1007/978-1-0716-3670-1_14.
Structural analysis of O-glycans from mucins and characterization of the interaction of these glycans with other biomolecules are essential for a full understanding of mucins. Various techniques have been developed for the structural and functional analysis of glycans. While 9-fluorenylmethyl chloroformate (Fmoc-Cl) is generally used to protect amino groups in peptide synthesis, it can also be used as a glycan-labeling reagent for structural analysis. Fmoc-labeled glycans are strongly fluorescent and can be analyzed with high sensitivity using liquid chromatography-fluorescence detection (LC-FD) analysis as well as being analyzed with high sensitivity by matrix-assisted laser desorption/ionization-time of flight mass spectrometry (MALDI-TOF MS). Fmoc-labeled glycans can be easily delabeled and converted to glycosylamine-form or free (hemiacetal or aldehyde)-form glycans that can be used to fabricate glycan arrays or synthesize glycosyl dendrimers. This derivatization allows for the isolation from biological samples of glycans that are difficult to synthesize chemically, as well as the fabrication of immobilized-glycan devices. The Fmoc labeling method promises to be a tool for accelerating O-glycan structural analysis and an understanding of molecular interactions. In this chapter, we introduce the Fmoc labeling method for analysis of O-glycans and fabrication of O-glycan arrays.
糖蛋白中 O-聚糖的结构分析以及这些聚糖与其他生物分子相互作用的特性对于充分了解糖蛋白至关重要。已经开发了各种技术来进行聚糖的结构和功能分析。虽然 9-芴甲氧羰基氯甲酸酯（Fmoc-Cl）通常用于保护肽合成中的氨基，但它也可用作结构分析的聚糖标记试剂。Fmoc 标记的聚糖具有很强的荧光性，可以通过液相色谱-荧光检测（LC-FD）分析进行高灵敏度分析，也可以通过基质辅助激光解吸/电离飞行时间质谱（MALDI-TOF MS）进行高灵敏度分析。Fmoc 标记的聚糖可以很容易地去标记并转化为糖苷胺形式或游离（半缩醛或醛）形式的聚糖，这些聚糖可用于制造糖基阵列或合成糖基树枝状聚合物。这种衍生化允许从生物样品中分离出难以化学合成的聚糖，并制造固定化聚糖装置。Fmoc 标记方法有望成为加速 O-聚糖结构分析和理解分子相互作用的工具。在本章中，我们介绍了用于分析 O-聚糖和制造 O-聚糖阵列的 Fmoc 标记方法。
