Capturing Protein-Protein Interactions with Acidic Amino Acids Reactive Cross-Linkers

文献类型： 外文期刊

第一作者： Liao, Qing-Qing

作者： Liao, Qing-Qing;Shu, Xin;Sun, Wei;Zhang, Long;Yang, Bing;Liao, Qing-Qing;Xie, Feng;Zhang, Zhengkui;Dai, Tong;Wang, Shuai;Zhou, Fangfang;Mandapaka, Hyma;Wu, Haifan;Zhao, Jinghua;Lin, Jinzhong;Jiang, Hong;Li, Shu-Wei;Coin, Irene;Yang, Fan;Peng, Jinrong;Li, Kui

作者机构：

关键词： acidic amino acid cross-linking; chemical cross-linker; cross-linking mass spectrometry; p53

期刊名称：SMALL （ 影响因子：13.3； 五年影响因子：13.2 ）

ISSN： 1613-6810

年卷期： 2023 年

页码：

收录情况： SCI

摘要： Acidic residues (Asp and Glu) have a high prevalence on protein surfaces, but cross-linking reactions targeting these residues are limited. Existing methods either require high-concentration coupling reagents or have low structural compatibility. Here a previously reported "plant-and-cast" strategy is extended to develop heterobifunctional cross-linkers. These cross-linkers first react rapidly with Lys sidechains and then react with Asp and Glu sidechains, in a proximity-enhanced fashion. The cross-linking reaction proceeds at neutral pH and room temperature without coupling reagents. The efficiency and robustness of cross-linking using model proteins, ranging from small monomeric proteins to large protein complexes are demonstrated. Importantly, it is shown that this type of cross-linkers are efficient at identifying protein-protein interactions involving acidic domains. The Cross-linking mass spectrometry (XL-MS) study with p53 identified 87 putative binders of the C-terminal domain of p53. Among them, SARNP, ZRAB2, and WBP11 are shown to regulate the expression and alternative splicing of p53 target genes. Thus, these carboxylate-reactive cross-linkers will further expand the power of XL-MS in the analysis of protein structures and protein-protein interactions. Acidic amino acids Asp/Glu widely distribute on protein surfaces and are involved in protein electrostatic interaction. In this work, stable and highly efficient acidic amino acids reactive cross-linkers are synthesized and applied to capture and identify p53 c-terminal domain-mediated direct protein interactions.image

分类号：

  • 相关文献
作者其他论文 更多>>

微信小程序