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TurboID 实现了蛋白质复合物和细胞类型特异性细胞器蛋白质组的邻近标记。

Proximity labeling of protein complexes and cell-type-specific organellar proteomes in enabled by TurboID.

机构信息

Department of Biology, Stanford University, Stanford, United States.

Howard Hughes Medical Institute, Chevy Chase, United States.

出版信息

Elife. 2019 Sep 19;8:e47864. doi: 10.7554/eLife.47864.

Abstract

Defining specific protein interactions and spatially or temporally restricted local proteomes improves our understanding of all cellular processes, but obtaining such data is challenging, especially for rare proteins, cell types, or events. Proximity labeling enables discovery of protein neighborhoods defining functional complexes and/or organellar protein compositions. Recent technological improvements, namely two highly active biotin ligase variants (TurboID and miniTurbo), allowed us to address two challenging questions in plants: (1) what are in vivo partners of a low abundant key developmental transcription factor and (2) what is the nuclear proteome of a rare cell type? Proteins identified with FAMA-TurboID include known interactors of this stomatal transcription factor and novel proteins that could facilitate its activator and repressor functions. Directing TurboID to stomatal nuclei enabled purification of cell type- and subcellular compartment-specific proteins. Broad tests of TurboID and miniTurbo in and and versatile vectors enable customization by plant researchers.

摘要

确定特定的蛋白质相互作用和空间或时间限制的局部蛋白质组有助于我们理解所有的细胞过程,但获得这些数据具有挑战性,特别是对于稀有蛋白质、细胞类型或事件。邻近标记可发现定义功能复合物和/或细胞器蛋白质组成的蛋白质邻域。最近的技术改进,即两种高活性的生物素连接酶变体(TurboID 和 miniTurbo),使我们能够解决植物中的两个具有挑战性的问题:(1)低丰度关键发育转录因子的体内伙伴是什么,以及(2)稀有细胞类型的核蛋白质组是什么?使用 FAMA-TurboID 鉴定的蛋白质包括这个气孔转录因子的已知相互作用蛋白和新的蛋白,这些蛋白可能有助于其激活剂和抑制剂功能。将 TurboID 引导至气孔核中,可纯化细胞类型和亚细胞区室特异性蛋白。TurboID 和 miniTurbo 在 和 中的广泛测试以及多功能载体,使植物研究人员能够进行定制。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/a19c/6791687/ce5dbda170d5/elife-47864-fig1.jpg

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