A photo-cross-linking approach to monitor protein dynamics in living cells
A photo-cross-linking approach to monitor protein dynamics in living cells
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DOI：
10.1016/j.bbagen.2019.03.003
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发表时间：
2020-02-01
影响因子：
3
通讯作者：
Mori, Hiroyuki
中科院分区：
文献类型：
--
作者：
Miyazaki, Ryoji;Akiyama, Yoshinori;Mori, Hiroyuki
背景：蛋白质是构成细胞的主要生物分子之一，在其生物发生和功能状态中与其他细胞因子相互作用。研究蛋白质的静态和瞬时相互作用对了解蛋白质的生理作用和调控机制具有重要意义。然而，只有有限的方法可以在分子水平上分析活细胞中蛋白质的动态行为。位点定向的体内光交联方法是一种在活细胞中以高空间分辨率捕获蛋白质相互作用的优雅技术。综述范围：本文综述了体内光交联方法，包括其最近的应用和需要考虑的潜在问题。我们还介绍了一种新的基于体内光交联的技术（PiXie），以高时空分辨率研究蛋白质动力学。主要结论：体内光交联使我们能够以高空间分辨率捕获弱/瞬态蛋白质相互作用，并允许识别相互作用因素。此外，PiXie方法可用于监测活细胞中蛋白质的快速折叠/组装过程。一般意义：体内光交联是一种简单的方法，已被用于分析许多细胞蛋白质的动态相互作用。该系统最初是在大肠杆菌中开发的，现已扩展到各种生物体的研究，使其成为研究许多细胞过程中动态蛋白质相互作用的基本技术。这篇文章是Daisuke Kohda博士编辑的题为“可视化“活”蛋白质分子的主要新技术”的特刊的一部分。
Background: Proteins, which comprise one of the major classes of biomolecules that constitute a cell, interact with other cellular factors during both their biogenesis and functional states. Studying not only static but also transient interactions of proteins is important to understand their physiological roles and regulation mechanisms. However, only a limited number of methods are available to analyze the dynamic behaviors of proteins at the molecular level in a living cell. The site-directed in vivo photo-cross-linking approach is an elegant technique to capture protein interactions with high spatial resolution in a living cell.Scope of review: Here, we review the in vivo photo-cross-linking approach including its recent applications and the potential problems to be considered. We also introduce a new in vivo photo-cross-linking-based technique (PiXie) to study protein dynamics with high spatiotemporal resolution.Major conclusions: In vivo photo-cross-linking enables us to capture weak/transient protein interactions with high spatial resolution, and allows for identification of interacting factors. Moreover, the PiXie approach can be used to monitor rapid folding/assembly processes of proteins in living cells.General significance: In vivo photo-cross-linking is a simple method that has been used to analyze the dynamic interactions of many cellular proteins. Originally developed in Escherichia coli, this system has been extended to studies in various organisms, making it a fundamental technique for investigating dynamic protein interactions in many cellular processes. This article is part of a Special issue entitled "Novel major techniques for visualizing `live' protein molecules" edited by Dr. Daisuke Kohda.