Genetically encoding ε-N-methacryllysine into proteins in live cells

杨兵 研究员

浙江大学生命科学研究院

时间：2024年11月15日（星期五）15:30

地点：思明校区化学楼234会议室　　　

厦门大学谱学分析与仪器教育部重点实验室

2024年11月11日

报告人简介：

杨兵，浙江大学生命科学研究院研究员，博士生导师，致力于生物质谱技术的开发与应用研究，拥有20年以上的生物质谱相关研究经历。设计与合成了近百种小分子交联剂和交联非天然氨基酸，其中10种以上已发表，并且开发了相应的生物质谱分析技术以及多种自主知识产权分析软件，用于在活细胞中原位的鉴定蛋白质之间的直接相互作用。相关研究成果以通讯作者（含共同）发表在Nature、Nature Cell Biology、Nature Communications、Science Advances、PNAS等国际期刊，发表论文的总引用率超过5500次，单篇最高引用率超过600次。授权美国发明专利一项、中国发明专利一项，申请中国发明专利一项。参与国家重点研发项目一项，主持国家级科研项目四项，省级科研项目两项。

报告摘要：

Lysine acylation is a ubiquitous post-translational modification (PTM) that plays pivotal roles in various cellular processes, such as transcription, metabolism, protein location and folding. Thousands of lysine acylation sites have been identified based on advances in antibody enrichment strategies, highly sensitive mass spectrometry (MS) analysis and bioinformatics. However, only 27 lysine methacrylation (Kmea) sites were identified, and they are exclusively located on histones. It is hard to separate, purify and differentiate Kmea peptides or proteins from its structural isomer lysine crotonylation (Kcr) with general biochemical approaches. Here, for the first time, we identified Kmea sites on a non-histone protein, Cyclophillin A (CypA). To investigate the function of Kmea on CypA, we developed a general genetic code expansion approach to incorporate an unnatural amino acid (Uaa) ε-N-Methacryllysine (MeaK) into target proteins and identified interacting proteins of methacrylated CypA using affinity-purification MS. We found that Kmea at CypA site 125 regulated cellular redox homeostasis and HDAC1 is the regulator of Kmea on CypA.