Chen Rui, Tolbert Thomas J
Methods Mol Biol. 2011;751:343-55. doi: 10.1007/978-1-61779-151-2_22.
This chapter describes a rapid and efficient approach for the solid-phase synthesis of N-linked glycopeptides that utilizes on-resin glycosylamine coupling to produce N-linked glycosylation sites. In this method, the full-length nonglycosylated peptide is first synthesized on a solid-phase support using standard Fmoc chemistry. The glycosylation site is then introduced through an orthogonally protected 2-phenylisopropyl (PhiPr) aspartic acid (Asp) residue. After selective deprotection of the Asp residue, a high mannose type oligosaccharide glycosylamine is coupled on-resin to the free Asp side chain to form a N-glycosidic bond. Subsequent protecting group removal and peptide cleavage from the resin ultimately yields the desired glycopeptide. This strategy provides an effective route for conducting glycosylation reactions on a solid-phase support, simplifies the process of glycopeptide purification relative to solution-phase glycopeptide synthesis strategies, and enables the recovery of potentially valuable, un-reacted oligosaccharides. This approach has been applied to the solid-phase synthesis of the N-linked high mannose glycosylated form of peptide T (ASTTTNYT), a fragment of the HIV-1 envelope glycoprotein gp120.
本章描述了一种用于N-连接糖肽固相合成的快速高效方法，该方法利用树脂上糖基胺偶联来产生N-连接糖基化位点。在此方法中，首先使用标准的Fmoc化学在固相载体上合成全长非糖基化肽。然后通过正交保护的2-苯基异丙基（PhiPr）天冬氨酸（Asp）残基引入糖基化位点。在Asp残基选择性脱保护后，将高甘露糖型寡糖糖基胺在树脂上偶联到游离的Asp侧链上，形成N-糖苷键。随后从树脂上去除保护基团并裂解肽，最终得到所需的糖肽。该策略为在固相载体上进行糖基化反应提供了一条有效途径，相对于溶液相糖肽合成策略简化了糖肽纯化过程，并能够回收潜在有价值的未反应寡糖。此方法已应用于HIV-1包膜糖蛋白gp120片段肽T（ASTTTNYT）的N-连接高甘露糖糖基化形式的固相合成。
