Peptide Institute, Inc., Ibaraki, Osaka, 567-0085, Japan.
Graduate School of Pharmaceutical Sciences, Kyoto University, Sakyo-ku, Kyoto, 606-8501, Japan.
Angew Chem Int Ed Engl. 2018 Feb 19;57(8):2105-2109. doi: 10.1002/anie.201711546. Epub 2018 Jan 25.
A solubilizing Trt-K tag was developed for the effective chemical preparation of peptides/proteins with low solubility. The Trt-K tag comprises a hydrophilic oligo-Lys sequence and a trityl anchor, and can be selectively introduced to a side chain thiol of Cys of deprotected peptides/proteins with a trityl alcohol-type introducing reagent Trt(OH)-K under acidic conditions. Significantly, the ligation product in the reaction mixture of a thiol-additive-free native chemical ligation can be modified directly in a one-pot manner to facilitate the isolation of the product by high-performance liquid chromatography. Finally, the Trt-K tag can be readily removed with a standard trifluoroacetic acid cocktail. Using this easy-to-attach/detach tag-aided method, a hepatitis B virus capsid protein that is usually difficult to handle was synthesized successfully.
开发了一种可溶的 Trt-K 标签，用于有效制备低溶解度的肽/蛋白质。Trt-K 标签由亲水寡聚赖氨酸序列和三苯甲基锚组成，可在酸性条件下，用三苯甲醇型引入试剂 Trt(OH)-K 选择性地引入到脱保护的肽/蛋白质的 Cys 的侧链巯基上。重要的是，在无硫醇添加剂的天然化学连接反应混合物中的连接产物可以以一锅法直接进行修饰，从而通过高效液相色谱法方便地分离产物。最后，Trt-K 标签可以用标准的三氟乙酸混合物轻松去除。使用这种易于连接/去除的标签辅助方法，成功合成了通常难以处理的乙型肝炎病毒衣壳蛋白。
