Li Jiahui, Cui Zenghui, Fan Chaochao, Zhou Yifei, Ren Mengtian, Zhou Chuanzheng
State Key Laboratory of Elemento-Organic Chemistry, Frontiers Science Center for New Organic Matter, Department of Chemical Biology, College of Chemistry, Nankai University Tianjin 300071 China
Chem Sci. 2023 Sep 19;14(39):10884-10891. doi: 10.1039/d3sc03719c. eCollection 2023 Oct 11.
Covalent trapping of DNA-binding proteins 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.
DNA结合蛋白的共价捕获：光交联是研究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，这些中间体与附近的官能团迅速且非选择性地反应，导致低的目标捕获产率和高的非目标背景捕获。在此，我们报道光笼蔽的2-丁烯-1,4-二醛（PBDA）是一种用于捕获DNA结合蛋白的高效光交联剂。对PBDA修饰的DNA进行光照射（360 nm）会产生2-丁烯-1,4-二醛（BDA），这是一种小的、寿命长的中间体，它与DNA结合蛋白的赖氨酸残基选择性反应，几分钟内即可形成产率高达70%的稳定DNA-蛋白质交联。此外，BDA对目标蛋白具有高特异性，导致非目标背景捕获率低。高光交联产率和目标特异性使PBDA成为研究DNA-蛋白质相互作用的有力工具。
