Yan Qibo, Li Ming, Zhang Yanxin, Liu Hailong, Liu Feng, Liao Weiwei, Wang Yingwu, Duan Haifeng, Wei Zhonglin
College of Chemistry, Jilin University, Changchun, 130012, Jilin, China.
Division of Chemical Metrology & Analytical Science, National Institute of Metrology, Beijing, 100029, China.
Talanta. 2023 Jun 1;258:124421. doi: 10.1016/j.talanta.2023.124421. Epub 2023 Mar 10.
Chemical cross-linking mass spectrometry (XL-MS) significantly contributes to the analysis of protein structures and the elucidation of protein-protein interactions. Currently available cross-linkers mainly target N-terminus, lysine, glutamate, aspartate, and cysteine residues in protein. Herein, a bifunctional cross-linker, named [4,4'-(disulfanediylbis(ethane-2,1-diyl)) bis(1-methyl-1,2,4-triazolidine-3,5-dione)] (DBMT) has been designed and characterized aiming to extremely expand the application of XL-MS approach. DBMT is capable of selectively targeting tyrosine residue in protein via an electrochemical click reaction, and/or targeting histidine residue in protein in the presence of O generated under photocatalytic reaction. A novel cross-linking strategy based on this cross-linker has been developed and demonstrated using model proteins, which provides a complementary XL-MS tool analyzing protein structure, protein complexes, protein-protein interactions, and even protein dynamics.
化学交联质谱法（XL-MS）对蛋白质结构分析和蛋白质-蛋白质相互作用的阐明有重大贡献。目前可用的交联剂主要针对蛋白质中的N端、赖氨酸、谷氨酸、天冬氨酸和半胱氨酸残基。在此，一种名为[4,4'-(二硫烷二基双(乙烷-2,1-二基))双(1-甲基-1,2,4-三唑烷-3,5-二酮)]（DBMT）的双功能交联剂已被设计和表征，旨在极大地扩展XL-MS方法的应用。DBMT能够通过电化学点击反应选择性地靶向蛋白质中的酪氨酸残基，和/或在光催化反应产生的O存在下靶向蛋白质中的组氨酸残基。基于这种交联剂开发了一种新的交联策略，并使用模型蛋白进行了验证，该策略为分析蛋白质结构、蛋白质复合物、蛋白质-蛋白质相互作用甚至蛋白质动力学提供了一种互补的XL-MS工具。
