Proximity labelling of interacting proteins by TurboID

Proximity labelling is a today commonly used biotin-based labelling method used to investigate protein-protein interactions in vivo and in vitro. Recently, TurboID was developed as an advanced version of the original proximity labelling protocols which allows better to analyse even small proteins in plants, as plants have a high background of endogenous biotin. The methodology, including detailed protocols for sample preparation and enrichment, gel electrophoresis, and immunodetection, as well as the necessary materials and equipment are shown in this chapter. It highlights the significance of TurboID in elucidating protein interaction networks within cells, including its application in studying organelle-specific proteins. Despite some limitations, such as potential non-specific labelling, the technique offers acquiring robust data that contribute to a deeper understanding of cellular processes. Future directions for research include refining the specificity of TurboID and expanding its use across various cell types and organisms.