Gao Ningguo
Department of Pharmacology, University of Texas, Southern Medical Center, Dallas, TX, USA.
Methods Mol Biol. 2006;347:1-12. doi: 10.1385/1-59745-167-3:1.
Defects in the synthesis of dolichol-linked oligosaccharide (or lipid-linked oligosaccharide [LLO]) cause severe, multisystem human diseases called type 1 congenital disorders of glycosylation (CDG type 1). LLOs are also involved in another disease, neuronal ceroid lipofuscinosis. Because of the low abundance of LLOs, almost all studies of LLO synthesis have relied upon metabolic labeling of the oligosaccharides with radioactive sugar precursors such as [3H]mannose or [14C]glucosamine, and therefore have been limited almost entirely to cell cultures and tissue slices. A procedure is presented for a facile, accurate, and sensitive non-radioactive method for LLO pathway analysis based on fluorophore-assisted carbohydrate electrophoresis (FACE). It is feasible to analyze almost any component in the LLO pathway with the application FACE, from sugar precursors to mature LLO (Glc3Man9GlcNAc2-P-P-dolichol).
多萜醇连接的寡糖（或脂连接寡糖[LLO]）合成缺陷会导致严重的多系统人类疾病，称为1型先天性糖基化障碍（1型CDG）。LLO还与另一种疾病——神经元蜡样脂褐质沉积症有关。由于LLO丰度较低，几乎所有关于LLO合成的研究都依赖于用放射性糖前体（如[3H]甘露糖或[14C]葡糖胺）对寡糖进行代谢标记，因此几乎完全局限于细胞培养和组织切片。本文介绍了一种基于荧光团辅助碳水化合物电泳（FACE）的简便、准确且灵敏的非放射性LLO途径分析方法。应用FACE分析LLO途径中的几乎任何成分，从糖前体到成熟的LLO（Glc3Man9GlcNAc2-P-P-多萜醇）都是可行的。
