组蛋白泛素化修饰参与调控DNA损伤修复、基因转录等重要生命过程，阐释泛素化修饰动态调控机制对相关疾病治疗开发具有重要意义。19F NMR能够实时动态分析不同时间尺度的蛋白质动力学信息，是研究蛋白质泛素化修饰动态调控机制的重要手段。本项目拟发展普适的氟代氨基酸合成方法，建立氟代氨基酸库；通过19F NMR开展组蛋白泛素化修饰对核小体的结构机制研究。具体内容为：a. 发展普适的氟代氨基酸合成方法，建立氟代氨基酸库；b. 建立合理的多肽或蛋白模型验证氟标氨基酸性能，掌握氟标氨基酸在多肽化学合成中的效率；c. 运用19F NMR技术探索泛素化修饰调控Dot1甲基转移酶活性机制，展示利用氟标非天然氨基酸通过核磁共振研究蛋白质动力学过程的优势。通过本项目的实施，有望建立一套结合19F标记蛋白质化学合成及核磁共振技术的泛素化修饰动态调控机制研究新策略。

Histone ubiquitination is involved in regulating numerous important life processes such as DNA damage repair, gene transcription, and so on. Interpretation of the dynamic regulation mechanism of ubiquitination is of guiding significance for the treatment of related diseases and drug development. 19F NMR can dynamically analyze the dynamic information of protein in different time scales, and is an important means to study the dynamic regulation mechanism of ubiquitination. In this project, we will develop general method for the synthesis of 19F labeled amino acids, and establish a 19F labeled amino acid library. With the chemical synthesis of 19F labeled protein, we will utilize 19F NMR to study the structural mechanism of histone ubiquitination on nucleosome. The specific content is: a. development of general method for the synthesis of 19F labeled amino acids and establishment of a 19F labeled amino acid library; b. establishment of protein model to verify the performance of 19F labeled amino acids and know well the efficiency of 19F labeled amino acids in the chemical synthesis of peptides; c. exploration of the mechanism of ubiquitination to regulate the activity of Dot1 methyltransferase, and demonstrate the advantages of 19F labeled non-natural amino acids using NMR to study the kinetic process of protein. Through the implementation of this project, it is expected to establish a new strategy for the study of the dynamic regulation mechanism of ubiquitination based on 19F labeled protein chemical synthesis and nuclear magnetic resonance technology.