Proximity-dependent labeling methods for proteomic profiling in living cells: An update.

Q1 Biochemistry, Genetics and Molecular Biology
Justin A Bosch, Chiao-Lin Chen, Norbert Perrimon
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引用次数: 46

Abstract

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.

活细胞蛋白质组学分析的邻近依赖标记方法:更新。
表征活细胞细胞器和亚细胞区域的蛋白质组组成可以促进对细胞组织以及蛋白质相互作用网络的理解。基于接近标记的方法与质谱(MS)相结合,为空间受限蛋白质组的系统分析提供了一种高通量方法。邻近标记利用酶产生活性自由基共价标记邻近的蛋白质。然后,标记的内源性蛋白可以被分离出来进行ms进一步分析。为了分析蛋白-蛋白相互作用或鉴定定位于离散亚细胞区室的成分,可以通过将酶融合到特定亚细胞区域的特定蛋白质或信号肽中来实现空间表达。虽然这些技术最近才被引入,但它们已经为广泛的生物过程提供了深刻的见解。在这里,我们提供了一个更新的描述和接近标记方法的比较,以及它们的应用和改进。由于每种方法都有其独特的特点,本综述的目的是描述如何使用不同的接近标记方法来回答不同的生物学问题。本文的分类为:技术>蛋白质分析。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
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来源期刊
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期刊介绍: Developmental biology is concerned with the fundamental question of how a single cell, the fertilized egg, ultimately produces a complex, fully patterned adult organism. This problem is studied on many different biological levels, from the molecular to the organismal. Developed in association with the Society for Developmental Biology, WIREs Developmental Biology will provide a unique interdisciplinary forum dedicated to fostering excellence in research and education and communicating key advances in this important field. The collaborative and integrative ethos of the WIREs model will facilitate connections to related disciplines such as genetics, systems biology, bioengineering, and psychology. The topical coverage of WIREs Developmental Biology includes: Establishment of Spatial and Temporal Patterns; Gene Expression and Transcriptional Hierarchies; Signaling Pathways; Early Embryonic Development; Invertebrate Organogenesis; Vertebrate Organogenesis; Nervous System Development; Birth Defects; Adult Stem Cells, Tissue Renewal and Regeneration; Cell Types and Issues Specific to Plants; Comparative Development and Evolution; and Technologies.
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