Membrane proteins have key functions in signal transduction, transport, and metabolism. Therefore, deciphering the interactions between membrane proteins provides crucial information on signal transduction and the spatiotemporal organization of protein complexes. However, detecting the interactions and behaviors of membrane proteins in their native environments remains difficult. Förster resonance energy transfer (FRET) is a powerful tool for quantifying the dynamic interactions and assembly of membrane proteins without disrupting their local environment, supplying nanometer-scale spatial information and nanosecond-scale temporal information. In this review, we briefly introduce the basic principles of FRET and assess the current state of progress in the development of new FRET techniques (such as FRET-FLIM, homo-FRET, and smFRET) for the analysis of plant membrane proteins. We also describe the various FRET-based biosensors used to quantify the homeostasis of signaling molecules and the active state of kinases. Furthermore, we summarize recent applications of these advanced FRET sensors in probing membrane protein interactions, stoichiometry, and protein clustering, which have shed light on the complex biological functions of membrane proteins in living plant cells.

中文翻译：

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通过 Förster 共振能量转移分析探测植物中的膜蛋白相互作用和信号分子稳态

膜蛋白在信号转导、转运和代谢中具有关键作用。因此，破译膜蛋白之间的相互作用提供了关于信号转导和蛋白质复合物时空组织的重要信息。然而，检测膜蛋白在其原生环境中的相互作用和行为仍然很困难。Förster 共振能量转移 (FRET) 是一种强大的工具，用于在不破坏其局部环境的情况下量化膜蛋白的动态相互作用和组装，提供纳米级空间信息和纳秒级时间信息。在这篇综述中，我们简要介绍了 FRET 的基本原理，并评估了新 FRET 技术（如 FRET-FLIM、homo-FRET、和 smFRET) 用于分析植物膜蛋白。我们还描述了各种基于 FRET 的生物传感器，用于量化信号分子的稳态和激酶的活性状态。此外，我们总结了这些先进的 FRET 传感器在探测膜蛋白相互作用、化学计量和蛋白质聚类方面的最新应用，这些应用揭示了活植物细胞中膜蛋白的复杂生物学功能。