Proximity-dependent labeling methods for proteomic profiling in living cells.
Proximity-dependent labeling methods for proteomic profiling in living cells.
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DOI：
10.1002/wdev.272
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发表时间：
2017-07
期刊：
影响因子：
--
通讯作者：
Perrimon N
中科院分区：
文献类型：
--
作者：
Chen CL;Perrimon N
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蛋白与生物素。生物素化的内源性蛋白可以通过质谱分离进一步分析。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，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，我们描述和比较当前的接近标记方法及其应用。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。
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 with biotin. The biotinylated 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 describe and compare current methods of proximity labeling as well as their applications. 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.
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影响因子：
6.4
作者：
Chen AL;Kim EW;Toh JY;Vashisht AA;Rashoff AQ;Van C;Huang AS;Moon AS;Bell HN;Bentolila LA;Wohlschlegel JA;Bradley PJ
通讯作者：
Bradley PJ
影响因子：
14.8
作者：
Hung V;Udeshi ND;Lam SS;Loh KH;Cox KJ;Pedram K;Carr SA;Ting AY
通讯作者：
Ting AY
影响因子：
8
作者：
Choi-Rhee, E;Schulman, H;Cronan, JE
通讯作者：
Cronan, JE
影响因子：
5.6
作者：
Cronan, JE
通讯作者：
Cronan, JE
影响因子：
4.7
作者：
Iwamaru, Yoshifumi;Kitani, Hiroshi;Yokoyama, Takashi
通讯作者：
Yokoyama, Takashi