邻近标记技术在蛋白质-蛋白质相互作用研究中的应用和进展

doi:10.16288/j.yczz.24-307

遗传 ›› 2025, Vol. 47 ›› Issue (6): 625-635.doi: 10.16288/j.yczz.24-307

邻近标记技术在蛋白质-蛋白质相互作用研究中的应用和进展

许昕薷¹(), 仇本华²^,³(), 鄢秀敏¹()

1.上海交通大学医学院附属新华医院环境与儿童健康重点实验室，上海 200092
2.中国科学院分子细胞科学卓越创新中心(生物化学与细胞生物学研究所)，细胞生物学国家重点实验室，上海 200031
3.中国科学院大学，北京 100049

收稿日期:2024-11-18 修回日期:2024-12-23 出版日期:2025-06-20 发布日期:2025-01-10
通讯作者: 鄢秀敏，博士，教授，研究方向：纤毛发生与功能。E-mail: yanx@shsmu.edu.cn
作者简介:许昕薷，博士研究生，研究方向：纤毛发生与功能。E-mail: 9894678@sjtu.edu.cn
仇本华，博士研究生，研究方向：纤毛发生与功能。E-mail: qiubenhua2018@sibcb.ac.cn
第一联系人：
许昕薷和仇本华并列第一作者。
基金资助:
基金项目：国家自然科学基金(32470724)

Application and progress of proximity labeling technology in the study of protein-protein interactions

Xinru Xu¹(), Benhua Qiu²^,³(), Xiumin Yan¹()

1. Ministry of Education-Shanghai Key Laboratory of Children’s Environmental Health, Xinhua Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai 200092, China
2. Key Laboratory of Multi-Cell Systems, Shanghai Institute of Biochemistry and Cell Biology, Center for Excellence in Molecular Cell Science, Chinese Academy of Sciences, Shanghai 200031, China
3. University of Chinese Academy of Sciences, Beijing 100049, China

Received:2024-11-18 Revised:2024-12-23 Published:2025-06-20 Online:2025-01-10
Supported by:
National Natural Science Foundation of China(32470724)

摘要/Abstract

摘要：

蛋白质-蛋白质相互作用在细胞内的几乎所有生物过程中都扮演着至关重要的角色。近年来，邻近标记技术作为一种前沿实验方法，被广泛应用于细胞内蛋白质相互作用和亚细胞区室空间蛋白质组等方面的研究。这项技术通过在目标蛋白质周围引入化学标记物，实现对邻近蛋白质的标记和捕捉。结合质谱鉴定与分析，邻近标记技术能够以高时空分辨率捕捉蛋白质间的相互作用及细胞区室的蛋白质组信息。与传统方法相比，邻近标记技术凭借其高时空分辨率和高灵敏度，成为研究细胞内蛋白质功能、相互作用及蛋白质空间组学的重要工具。本文系统介绍了蛋白质邻近标记技术的原理、分类及其应用，以期为研究人员提供全面的技术视角，促进蛋白质邻近标记技术在细胞生物学、蛋白质组学以及疾病机制研究中的更广泛应用。

关键词: 邻近标记技术, 蛋白质相互作用, 空间蛋白质组

Abstract:

Protein-protein interactions are essential to almost all biological processes within cells. In recent years, proximity labeling technology has emerged as a cutting-edge approach for studying protein interactions and subcellular compartment proteomics. This technique enables labeling and capture of neighboring proteins by introducing chemical tags in the vicinity of a target protein. When combined with mass spectrometry, proximity labeling facilitates the effective identification of protein-protein interactions and proteomic landscapes of subcellular compartments with high spatiotemporal resolution. Compared to traditional methods, proximity labeling offers superior resolution, sensitivity, and real-time tracking capabilities，making it a powerful tool for investigating protein functions, interactions, and spatial proteomics within cells. In this review, we provide a comprehensive overview of the principles, classifications, and applications of proximity labeling technology, aiming to offer researchers valuable insights into this innovative methodology and to promote its broader application in cell biology, proteomics, and the study of disease mechanisms.

Key words: proximity labeling technology, protein-protein interactions, spatial proteome

许昕薷, 仇本华, 鄢秀敏. 邻近标记技术在蛋白质-蛋白质相互作用研究中的应用和进展[J]. 遗传, 2025, 47(6): 625-635.

Xinru Xu, Benhua Qiu, Xiumin Yan. Application and progress of proximity labeling technology in the study of protein-protein interactions[J]. Hereditas(Beijing), 2025, 47(6): 625-635.

图/表 2

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方法	基于生物素连接酶的邻近标记技术			基于过氧化物酶的邻近标记技术
方法	BioID	TurboID	miniTurboID	HRP	APEX	APEX2
分子量大小	35 kDa	35 kDa	28 kDa	44 kDa	27 kDa	27 kDa
特点	BirA来自大肠杆菌，BioID基于BirA的R118G突变体(BirA*)	改良型BirA*，通过15个点突变提高催化效率，标记范围约为10~20 nm，标记时间为10 min	TurboID的改良版本，删除了N端区域并只携带有13个点突变，小分子量减少对目标蛋白功能和定位的干扰	酶催化活性受pH值的影响，在不同的细胞区室中酶催化活性有所不同	APX是一种植物酶，经过工程化改造的APEX，催化活性高	APEX的A134P突变体，具有更高的催化活性和稳定性
是否需要H₂O₂	否			是
反应底物	Biotin			Biotin-phenol
反应时间	18~24 h	10 min	10 min	5~10 min	≤1 min	≤1 min
标记范围	10 nm	10~20 nm	10~20 nm	200~300 nm	20 nm	20 nm
背景噪音	中等	中等	低	高	中等	中等
应用	用于识别邻近蛋白，适合研究细胞内相互作用	捕获瞬时和动态蛋白质相互作用，应用于多种模式生物		用于标记邻近蛋白，但应用受到细胞环境限制	用于空间蛋白质组和蛋白质相互作用研究以及电镜成像

邻近标记技术在蛋白质-蛋白质相互作用研究中的应用和进展

Application and progress of proximity labeling technology in the study of protein-protein interactions

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