The First Affiliated Hospital of USTC, Division of Life Sciences and Medicine, Hefei National Laboratory for Physical Sciences at the Microscale, University of Science and Technology of China, Hefei, Anhui, China.
Methods Mol Biol. 2022;2530:241-256. doi: 10.1007/978-1-0716-2489-0_16.
Chemical synthesis can provide hydrophobic proteins with natural or man-made modifications (e.g. S-palmitoylation, site-specific isotope labeling and mirror-image proteins) that are difficult to obtain through the recombinant expression technology. The difficulty of chemical synthesis of hydrophobic proteins stems from the hydrophobic nature. Removable backbone modificaiton (RBM) strategy has been developed for solubilizing the hydrophobic peptides/proteins. Here we take the chemical synthesis of a S-palmitoylated peptide as an example to describe the detailed procedure of RBM strategy. Three critical steps of this protocol are: (1) installation of Lys6-tagged RBM groups into the peptides by Fmoc (9-fluorenylmethyloxycarbonyl) solid-phase peptide synthesis, (2) chemical ligation of the peptides, and (3) removal of the RBM tags by TFA (trifluoroacetic acid) cocktails to give the target peptide.
化学合成可以为疏水性蛋白质提供天然或人工修饰（例如 S-棕榈酰化、定点同位素标记和镜像蛋白质），这些修饰很难通过重组表达技术获得。疏水性蛋白质的化学合成困难源于其疏水性。可去除骨架修饰（RBM）策略已被开发用于溶解疏水性肽/蛋白质。在这里，我们以 S-棕榈酰化肽的化学合成为例，描述 RBM 策略的详细步骤。该方案有三个关键步骤：（1）通过 Fmoc（9-芴甲氧羰基）固相肽合成将带有 Lys6 标记的 RBM 基团安装到肽中，（2）肽的化学连接，以及（3）通过 TFA（三氟乙酸）混合物去除 RBM 标记以得到目标肽。
