Department of Biochemistry & Research Center for Emerging Infections and Zoonoses (RIZ), University of Veterinary Medicine Hannover, Hannover, Germany; Institute for Experimental Virology, TWINCORE, Centre for Experimental and Clinical Infection Research, a Joint Venture Between the Medical School Hannover and the Helmholtz Centre for Infection Research, Hannover, Germany; Department of Clinical Microbiology, Virology, Umeå University, Umeå, Sweden; Wallenberg Centre for Molecular Medicine (WCMM), Umeå University, Umeå, Sweden.
Department of Biochemistry & Research Center for Emerging Infections and Zoonoses (RIZ), University of Veterinary Medicine Hannover, Hannover, Germany; Institute for Experimental Virology, TWINCORE, Centre for Experimental and Clinical Infection Research, a Joint Venture Between the Medical School Hannover and the Helmholtz Centre for Infection Research, Hannover, Germany.
Adv Virus Res. 2021;109:63-104. doi: 10.1016/bs.aivir.2021.02.001. Epub 2021 Apr 16.
Cellular compartmentalization of proteins and protein complex formation allow cells to tightly control biological processes. Therefore, understanding the subcellular localization and interactions of a specific protein is crucial to uncover its biological function. The advent of proximity labeling (PL) has reshaped cellular proteomics in infection biology. PL utilizes a genetically modified enzyme that generates a "labeling cloud" by covalently labeling proteins in close proximity to the enzyme. Fusion of a PL enzyme to a specific antibody or a "bait" protein of interest in combination with affinity enrichment mass spectrometry (AE-MS) enables the isolation and identification of the cellular proximity proteome, or proxisome. This powerful methodology has been paramount for the mapping of membrane or membraneless organelles as well as for the understanding of hard-to-purify protein complexes, such as those of transmembrane proteins. Unsurprisingly, more and more infection biology research groups have recognized the potential of PL for the identification of host-pathogen interactions. In this chapter, we introduce the enzymes commonly used for PL labeling as well as recent promising advancements and summarize the major achievements in organelle mapping and nucleic acid PL. Moreover, we comprehensively describe the research on host-pathogen interactions using PL, giving special attention to studies in the field of virology.
蛋白质的细胞区室化和蛋白质复合物的形成允许细胞严格控制生物过程。因此，了解特定蛋白质的亚细胞定位和相互作用对于揭示其生物学功能至关重要。邻近标记 (PL) 的出现改变了感染生物学中的细胞蛋白质组学。PL 利用一种经过基因改造的酶，通过共价标记酶附近的蛋白质来产生“标记云”。将 PL 酶融合到特定的抗体或感兴趣的“诱饵”蛋白上，并结合亲和富集质谱 (AE-MS)，可用于分离和鉴定细胞邻近蛋白质组，或邻近蛋白质组。这种强大的方法对于膜或无膜细胞器的映射以及难以纯化的蛋白质复合物（如跨膜蛋白复合物）的理解至关重要。毫不奇怪，越来越多的感染生物学研究小组已经认识到 PL 用于鉴定宿主-病原体相互作用的潜力。在本章中，我们介绍了常用于 PL 标记的酶，以及最近有前途的进展，并总结了在细胞器映射和核酸 PL 方面的主要成就。此外，我们全面描述了使用 PL 进行宿主-病原体相互作用的研究，特别关注病毒学领域的研究。
