Detection and Quantification of Biotinylated Proteins for TurboID-Based Proximity Labeling Mass Spectrometry in...

TurboID-based proximity labeling, combined with mass spectrometry (PL-MS), has been widely adopted for studies in both plants and animals for a variety of applications. In this approach, an engineered biotin ligase, TurboID, fused to a bait protein, biotinylates proteins in close proximity to the bait protein following biotin treatment, allowing their subsequent isolation using streptavidin beads and identification by mass spectrometry. This chapter provides a detailed step-by-step protocol for sample preparation using Arabidopsis tissues, along with an overview of data analysis strategies and software tools for the reliable identification and quantification of biotinylated proteins.