Covalent trapping of DNA-binding proteins via photo-crosslinking is an advantageous method for studying DNA–protein interactions. However, traditional photo-crosslinkers generate highly reactive intermediates that rapidly and non-selectively react with nearby functional groups, resulting in low target-capture yields and high non-target background capture. Herein, we report that photo-caged 2-butene-1,4-dial (PBDA) is an efficient photo-crosslinker for trapping DNA-binding proteins. Photo-irradiation (360 nm) of PBDA-modified DNA generates 2-butene-1,4-dial (BDA), a small, long-lived intermediate that reacts selectively with Lys residues of DNA-binding proteins, leading in minutes to stable DNA–protein crosslinks in up to 70% yield. In addition, BDA exhibits high specificity for target proteins, leading to low non-target background capture. The high photo-crosslinking yield and target specificity make PBDA a powerful tool for studying DNA–protein interactions.

中文翻译：

---

光笼 2-丁烯-1,4-dial 作为一种有效的、目标特异性的光交联剂，用于共价捕获 DNA 结合蛋白

通过光交联共价捕获 DNA 结合蛋白是研究 DNA-蛋白质相互作用的一种有利方法。然而，传统的光交联剂会产生高反应性中间体，这些中间体会与附近的官能团快速且非选择性地发生反应，导致目标捕获率低和非目标背景捕获率高。在此，我们报告光笼 2-丁烯-1,4-拨号 (PBDA) 是一种有效的光交联剂，用于捕获 DNA 结合蛋白。PBDA 修饰的 DNA 经光照射 (360 nm) 生成 2-丁烯-1,4-二旋体 (BDA)，这是一种小型、长寿命的中间体，可选择性地与 DNA 结合蛋白的赖氨酸残基反应，在几分钟内形成稳定的产物。 DNA-蛋白质交联率高达 70%。此外，BDA 对目标蛋白表现出高度特异性，导致非目标背景捕获率较低。高光交联产率和目标特异性使 PBDA 成为研究 DNA-蛋白质相互作用的强大工具。