Højlys-Larsen Kim B, Jensen Knud J
Symphogen A/S, Lyngby, Denmark.
Methods Mol Biol. 2013;1047:191-9. doi: 10.1007/978-1-62703-544-6_13.
Phosphopeptides are generally prepared by incorporation of suitable, protected phosphoamino acid derivatives during peptide synthesis using routine coupling protocols. The feasibility of chemical synthesis of phosphorylated peptides by Fmoc-SPPS was greatly enhanced by the introduction of the monobenzyl protecting group for the phosphate group. This minimized β-elimination of the phosphate group and made Fmoc-based synthesis of phosphopeptides the preferred synthesis strategy. Described here is our strategy for the synthesis of phosphopeptides attached to the solid support PEGA via a backbone amide linker type. This linker allows removal of side-chain protection groups without releasing the phosphopeptide from the solid support, thus enabling solid-phase-based pull-down reactions and peptide-protein interaction studies.
磷酸化肽通常是在肽合成过程中，使用常规偶联方案引入合适的、受保护的磷酸氨基酸衍生物来制备的。通过引入用于磷酸基团的单苄基保护基团，Fmoc固相肽合成法（Fmoc-SPPS）化学合成磷酸化肽的可行性得到了极大提高。这将磷酸基团的β消除降至最低，并使基于Fmoc的磷酸化肽合成成为首选的合成策略。本文描述了我们通过主链酰胺连接体类型合成连接到固相载体PEGA上的磷酸化肽的策略。这种连接体允许去除侧链保护基团，而不会使磷酸化肽从固相载体上释放，从而能够进行基于固相的下拉反应和肽-蛋白质相互作用研究。
