项目名称：面向蛋白甲基化的智能富集新材料
项目来源：国家自然科学基金面上项目
项目编号：21775116
项目金额：65万元（直接经费）
蛋白甲基化是最常见的翻译后修饰方式之一，在表观遗传调控中发挥着至关重要的作用。然而，甲基化形式的多样性、低丰度和生物样品的复杂性，导致在蛋白质组学水平上捕获、分析、鉴定甲基化位点非常困难。因此，急需开发对各种类型的甲基化肽链具有高效捕获能力的人工材料和富集方法。在本项目中，申请人创新地利用生物分子响应性聚合物的设计思想，尝试解决甲基化肽链的富集和分离挑战。借助模仿甲基化位点结合蛋白结构域中的氨基酸序列，构筑合适的短肽，实现对甲基化肽的精确识别。进而将这些短肽与各种功能单体共聚，制备智能聚合物材料。通过调节溶剂极性、温度、pH值、离子强度等参数，实现甲基化肽段在聚合物表面上动态可调的吸附、脱附过程。在此基础上建立富集方法学，评估材料在组蛋白模型样品、细胞、组织样本中对甲基化肽的富集性能。智能聚合物结合力强、选择性高、可控性好、易洗脱、易扩展等特点，将为甲基化肽富集材料的开发提供一种新的思路。
Title of funded project：Smart Enrichment Materials for Protein Methylation
Funding agency：National Natural Science Foundation of China
Grant number：21775116
Total funding amount：CNY 650,000
Protein methylation is one of the most common and important post-translational modification types, which plays essential roles in epigenetic regulation. However, owing to the diversity of protein methylation, ultra-low abundance and the inherent complexity of biosamples, it is extremely difficult to capture, analysis and identify the methylation sites at the level of proteomics. Thus, developing high-efficiency enrichment materials and corresponding methods capable of capturing various types of methylated peptides is highly urgent, which could not be resolved by artificial materials by far. In this project, we innovatively introduce the design idea of biomolecule-responsive polymer to tackle the challenges of methylated peptide enrichment and separation. By mimicking the peptide sequences and the unique conformation of binding domains in methylation site-specific binding proteins, the optimized oligopeptide sequences could be designed for recognizing the methylation sites precisely. In addition, these oligopeptides would be copolymerized with diverse functional monomers to prepare smart polymer-based enrichment materials. Then dynamic adsorption or desorption behaviours of methylated peptides on these smart polymer surfaces could be modulated by various chromatographic conditions, such as solvent polarity, solution pH, temperature or ionic strength. Based on these researches, the optimized enrichment methodology could be built reasonably, and the enrichment efficiency of our materials towards methylated peptides will be evaluated in model histone protein, cell and tissue samples. The remarkable advantages of smart polymers in high binding affinity, satisfactory selectivity, good controllability, easy elution and expandable material platform, provide a new insight for the development of methylated peptide enrichment materials.