期刊
WILEY INTERDISCIPLINARY REVIEWS-DEVELOPMENTAL BIOLOGY
卷 10, 期 1, 页码 -出版社
WILEY
DOI: 10.1002/wdev.392
关键词
APEX; BioID; HRP; proximity labeling PUP-IT
资金
- Damon Runyon Foundation [2258-16]
- NIH [5R01DK121409]
Proteome composition of organelles and subcellular regions can be analyzed using proximity labeling-based methods coupled with mass spectrometry (MS) to understand cellular organization and protein interactome networks. These methods utilize enzymes that generate reactive radicals to covalently tag neighboring proteins, allowing for systematic analysis of spatially restricted proteomes. By fusing enzymes to specific proteins or signal peptides targeting to particular subcellular regions, spatial expression can be achieved for studying protein-protein interactions or identifying components in discrete subcellular compartments.
Characterizing the proteome composition of organelles and subcellular regions of living cells can facilitate the understanding of cellular organization as well as protein interactome networks. Proximity labeling-based methods coupled with mass spectrometry (MS) offer a high-throughput approach for systematic analysis of spatially restricted proteomes. Proximity labeling utilizes enzymes that generate reactive radicals to covalently tag neighboring proteins. The tagged endogenous proteins can then be isolated for further analysis by MS. To analyze protein-protein interactions or identify components that localize to discrete subcellular compartments, spatial expression is achieved by fusing the enzyme to specific proteins or signal peptides that target to particular subcellular regions. Although these technologies have only been introduced recently, they have already provided deep insights into a wide range of biological processes. Here, we provide an updated description and comparison of proximity labeling methods, as well as their applications and improvements. As each method has its own unique features, the goal of this review is to describe how different proximity labeling methods can be used to answer different biological questions. This article is categorized under: Technologies > Analysis of Proteins
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