Ligation of Soluble but Unreactive Peptide Segments in the Chemical Synthesis of Haemophilus Influenzae DNA Ligase
Ligation of Soluble but Unreactive Peptide Segments in the Chemical Synthesis of Haemophilus Influenzae DNA Ligase
复制标题
化学合成流感嗜血杆菌 DNA 连接酶中可溶但不反应的肽段的连接
DOI：
10.1002/anie.201905149
复制
发表时间：
2019
期刊：
影响因子：
--
通讯作者：
Liu Lei
中科院分区：
文献类型：
--
作者：
Zhang Baochang;Deng Qiang;Zuo Chong;Yan Bingjia;Zuo Chao;Cao Xiu Xiu;Zhu Ting F;Zheng Ji Shen;Liu Lei
在水溶性球状流感嗜血杆菌DNA连接酶（Hin-Lig）的全化学合成过程中，我们观察到可溶性肽段未能进行天然化学连接的令人惊讶的现象。基于动态光散射和透射电子显微镜实验，我们确定该肽在含有6m盐酸胍的均匀溶液中形成可溶性胶体颗粒。传统的肽性能改进策略，例如安装末端/侧链 Arg 标签或 O-酰基异肽，未能实现该反应，可能是因为它们无法破坏可溶性胶体颗粒的形成。然而，最近开发的用于合成膜蛋白的可移除主链修饰策略确实破坏了胶体的形成，并且最终实现了这种可溶但不反应的系统的所需连接。这项工作表明，除了良好的肽溶解度之外，适当的溶液分散状态也是肽连接成功的先决条件。
During the total chemical synthesis of the water‐soluble globularHaemophilus InfluenzaeDNA ligase (Hin‐Lig), we observed the surprising phenomenon of a soluble peptide segment that failed to undergo native chemical ligation. Based on dynamic light scattering and transmission electron microscopy experiments, we determined that the peptide formed soluble colloidal particles in a homogeneous solution containing 6mguanidine hydrochloride. Conventional peptide performance‐improving strategies, such as installation of a terminal/side‐chain Arg tag orO‐acyl isopeptide, failed to enable the reaction, presumably because of their inability to disrupt the formation of soluble colloidal particles. However, a removable backbone modification strategy recently developed for the synthesis of membrane proteins did disrupt the formation of the colloids, and the desired ligation of this soluble but unreactive system was eventually accomplished. This work demonstrates that an appropriate solution dispersion state, in addition to good peptide solubility, is a prerequisite for successful peptide ligation.