A wide range of technologies have been developed to study protein–protein interactions and to map comprehensive protein interaction networks. In recent years, advances in proximity labeling methods, coupled to protein identification by mass spectrometry, have led to exponential growth in the popularity of these methods. In this chapter, we present a detailed protocol using TurboID in Arabidopsis thaliana seedlings, using membrane-bound transcription factors as a case study. Additionally, we outline the workflow for analyzing mass spectrometry data to identify putative protein interactors.

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Characterization of the Protein Interactome of Membrane-Bound Transcription Factors Using TurboID-Based Proximity Labeling in Planta

  • Jonas De Backer,
  • Lindsy De Veirman,
  • Inge De Clercq

摘要

A wide range of technologies have been developed to study protein–protein interactions and to map comprehensive protein interaction networks. In recent years, advances in proximity labeling methods, coupled to protein identification by mass spectrometry, have led to exponential growth in the popularity of these methods. In this chapter, we present a detailed protocol using TurboID in Arabidopsis thaliana seedlings, using membrane-bound transcription factors as a case study. Additionally, we outline the workflow for analyzing mass spectrometry data to identify putative protein interactors.