Tuning the reactivity of a substrate for SNAP-tag expands its application for recognition-driven DNA-protein conjugation
Tuning the reactivity of a substrate for SNAP-tag expands its application for recognition-driven DNA-protein conjugation
复制标题
调整 SNAP 标签底物的反应性扩展了其在识别驱动的 DNA-蛋白质缀合方面的应用
DOI：
10.1002/chem.202103304
复制
发表时间：
2021
影响因子：
4.3
通讯作者：
Takashi Morii
中科院分区：
文献类型：
--
作者：
Zhengxiao Zhang;Eiji Nakata;Huyen Dinh;Masayuki Saimura;Arivazhagan Rajendran;Kazunari Matsuda;Takashi Morii
识别驱动的修饰已成为一种新颖的方法，可以特异性、高效地修饰感兴趣的生物分子靶标。为此，蛋白质模块化接头 (MA) 是识别驱动的 DNA 修饰的理想反应模型，因为它们由序列特异性 DNA 结合域 (DBD) 和自连接蛋白质标签组成。将 DBD 的 DNA 识别与蛋白质标签的化学选择性反应结合起来可以提供高效的序列特异性反应。然而，将由反应蛋白标签及其底物（例如 SNAP 标签和苄基鸟嘌呤 (BG)）组成的 MA 结合起来，显示出与 DNA 的非选择性反应。因此，设计了 SNAP 标签的新底物，以实现 MA 与 SNAP 标签的序列选择性快速交联反应。通过动力学分析验证了带有 SNAP 标签的底物的反应，以实现 MA 的序列选择性交联反应。新的底物使 SNAP 标签与 CLIP 标签具有独特的正交性，从而通过六个独特的 MA 实现正交 DNA-蛋白质交联。
Recognition‐driven modification has been emerging as a novel approach to modifying biomolecular targets of interest site‐specifically and efficiently. To this end, protein modular adaptors (MAs) are the ideal reaction model for recognition‐driven modification of DNA as they consist of both a sequence‐specific DNA‐binding domain (DBD) and a self‐ligating protein‐tag. Coupling DNA recognition by DBD and the chemoselective reaction of the protein tag could provide a highly efficient sequence‐specific reaction. However, combining an MA consisting of a reactive protein‐tag and its substrate, for example, SNAP‐tag and benzyl guanine (BG), revealed rather nonselective reaction with DNA. Therefore new substrates of SNAP‐tag have been designed to realize sequence‐selective rapid crosslinking reactions of MAs with SNAP‐tag. The reactions of substrates with SNAP‐tag were verified by kinetic analyses to enable the sequence‐selective crosslinking reaction of MA. The new substrate enables the distinctive orthogonality of SNAP‐tag against CLIP‐tag to achieve orthogonal DNA‐protein crosslinking by six unique MAs.