小柯机器人

近日，美国斯坦福大学Alice Y. Ting及其课题组通过TransitID对活细胞内和活细胞间的蛋白质组贩运进行动态测绘。2023年6月28日，《细胞》杂志在线发表了这项成果。

研究人员报告了TransitID，一种在活细胞中以纳米级空间分辨率无偏见地绘制内源性蛋白质组贩运图谱的方法。两种接近标记（PL）酶，TurboID和APEX，分别针对源区和目的区，通过依次加入它们的小分子底物，用每种酶串联进行PL。质谱分析确定了被两种酶标记的蛋白质。利用TransitID，研究人员绘制了细胞膜和线粒体、细胞膜和细胞核以及细胞核和应激颗粒（SG）之间的蛋白质组贩运图谱，发现了SG在保护转录因子JUN免受氧化应激中的作用。TransitID还确定了巨噬细胞和癌细胞之间的细胞间信号的蛋白质。TransitID提供了一个强大的方法来区分基于细胞区段或细胞类型的蛋白质群。

据悉，在活细胞中一次性绘制数千种内源性蛋白贩运图谱的能力将揭示目前显微镜和质谱法都无法看到的生物学特性。

附：英文原文

Title: Dynamic mapping of proteome trafficking within and between living cells by TransitID

Author: Wei Qin, Joleen S. Cheah, Charles Xu, James Messing, Brian D. Freibaum, Steven Boeynaems, J. Paul Taylor, Namrata D. Udeshi, Steven A. Carr, Alice Y. Ting

Issue&Volume: 2023-06-28

Abstract: The ability to map trafficking for thousands of endogenous proteins at once in livingcells would reveal biology currently invisible to both microscopy and mass spectrometry.Here, we report TransitID, a method for unbiased mapping of endogenous proteome traffickingwith nanometer spatial resolution in living cells. Two proximity labeling (PL) enzymes,TurboID and APEX, are targeted to source and destination compartments, and PL witheach enzyme is performed in tandem via sequential addition of their small-moleculesubstrates. Mass spectrometry identifies the proteins tagged by both enzymes. UsingTransitID, we mapped proteome trafficking between cytosol and mitochondria, cytosoland nucleus, and nucleolus and stress granules (SGs), uncovering a role for SGs inprotecting the transcription factor JUN from oxidative stress. TransitID also identifiesproteins that signal intercellularly between macrophages and cancer cells. TransitIDoffers a powerful approach for distinguishing protein populations based on compartmentor cell type of origin.

DOI: 10.1016/j.cell.2023.05.044

Source: https://www.cell.com/cell/fulltext/S0092-8674(23)00596-2