Park Tae-Ki, Kim Tae-Wuk
Department of Life Science, Hanyang University, Seoul 04763, Republic of Korea; Hanyang Institute of Bioscience and Biotechnology, Hanyang University, Seoul 04763, Republic of Korea.
Department of Life Science, Hanyang University, Seoul 04763, Republic of Korea; Hanyang Institute of Bioscience and Biotechnology, Hanyang University, Seoul 04763, Republic of Korea; Research Institute for Convergence of Basic Science, Hanyang University, Seoul 04763, Republic of Korea.
Mol Cells. 2025 Aug;48(8):100236. doi: 10.1016/j.mocell.2025.100236. Epub 2025 Jun 3.
Proximity labeling-mass spectrometry has emerged as a powerful biological tool for elucidating the intracellular protein interactome of target proteins. Here, we introduce experimental strategies and methodologies for understanding complex protein-protein interactions in plant cells using the recently developed TurboID-based PL approach. We provide a brief overview of the overall procedures, including the preparation of transgenic Arabidopsis plants, biotin treatment, affinity purification, on-bead digestion, liquid chromatography-tandem mass spectrometry, and data analysis.
邻近标记质谱法已成为一种强大的生物学工具,用于阐明目标蛋白的细胞内蛋白质相互作用组。在这里,我们介绍了使用最近开发的基于TurboID的邻近标记方法来理解植物细胞中复杂蛋白质-蛋白质相互作用的实验策略和方法。我们简要概述了整个流程,包括转基因拟南芥植物的制备、生物素处理、亲和纯化、珠上消化、液相色谱-串联质谱分析和数据分析。