适用于活性化合物靶点鉴定的无标记蛋白质分析方法  

Label-free Protein Analysis Methods for Active Compound Targets Identification

作　　者：吕博海 勾文峰 许飞飞 李艳丽 李祎亮[2] 侯文彬 Lyu Bohai;Gou Wenfeng;Xu Feifei;Li Yanli;Li Yiliang;Hou Wenbin(Institute of Traditional Chinese Medicine,Tianjin University of Traditional Chinese Medicine,Tianjin 301617;Institute of Radiation Medicine,Peking Union Medical College&Chinese Academy of Medical Sciences,Tianjin 300192)

机构地区：[1]天津中医药大学中医药研究院,天津301617 [2]北京协和医学院&中国医学科学院放射医学研究所,天津300192

出　　处：《化学学报》2024年第6期629-640,共12页Acta Chimica Sinica

基　　金：国家自然科学基金(Nos.82104012,82202950,82303681);中国医学科学院医学与健康科技创新工程重大协同创新项目(No.2021-I2M-1-042);中央高校基本科研业务费专项资金(No.3332022063)资助。

摘　　要：随着越来越多的活性有效成分被发现,探寻这些有效成分的药理作用机制,鉴定它们有效靶点的需求逐渐增加.化学生物学作为近年来新发展的一门交叉学科,是承担此项任务的最佳选择.其中多种无标记蛋白质分析方法通过研究活性分子化合物与蛋白质相互作用,从而影响蛋白的化学物理性质,为药物研发提供新的思路.这些方法通过检测活性化合物对蛋白质的热稳定性、酶敏感性和分子结构引起的改变,以及结合光谱、质谱分析等相关技术,来评估活性化合物的选择性和作用范围,减少脱靶风险.总结了多种适用于靶点鉴定的无标记蛋白质分析方法,对药物的靶点发现具有参考意义.As more and more active ingredients are discovered,there is an increasing demand to explore the pharmacologi-cal mechanisms of these drugs and identify their effective targets.Chemical biology,as a newly developed interdisciplinary field in recent years,is the best choice to undertake this task.The identification method of unlabeled targets does not involve any chemical modification of small molecular drugs and has attracted wide attention in recent years.Most of the target iden-tification methods based on interaction belong to the type of unlabeled deconvolution.Label-free protein analysis can also help predict potential drug targets or candidates,develop new biomarker assays and diagnostic reagents,and evaluate the selectivity and range of active compounds to reduce the risk of off-target effects.It can achieve these goals using techniques such as changing protein thermal stability,enzyme sensitivity,and molecular structure and using mass spectrometry.In this paper,we review the reported Label-free protein analysis techniques for identifying different types of targets.Based on dif-ferent principles from three perspectives,hydrogen deuterium exchange mass spectrometry,surface plasmon resonance,mi-croscale thermophoresis,fluorescence correlation spectroscopy,and other techniques are introduced,and the principles and scope of application of these techniques are introduced to readers through figures,texts,and tables.The combined use of various techniques can improve the success rate of target discovery,but this review still has the limitation of incomplete summary of the techniques.To give the reader an initial impression of Label-free protein analysis techniques that can provide valuable insights for drug target discovery.The target identification of active compounds can deepen our understanding of the mode of action of clinical drugs,help to discover new undruggable proteins,and provide the possibility for innovative treat-ments.With the development of mass spectrometry-based proteomics and computational biology,

关 键 词：靶点鉴定 无标记 蛋白质分析 热力学作用 配体结合 荧光报告 

分 类 号：Q503[生物学—生物化学] TQ460.72[医药卫生—药物分析学]