Chinese Bulletin of Botany ›› 2021, Vol. 56 ›› Issue (5): 584-593.DOI: 10.11983/CBB21104

• INVITED PROTOCOL • Previous Articles     Next Articles

Methods for TurboID-based Proximal Labeling in Plants

Jiayi Kuang, Hongqing Li, Wenjin Shen*(), Caiji Gao*()   

  1. Guangdong Provincial Key Laboratory of Biotechnology for Plant Development, School of Life Sciences, South China Normal University, Guangzhou 510631, China
  • Received:2021-07-01 Accepted:2021-08-09 Online:2021-09-01 Published:2021-08-31
  • Contact: Wenjin Shen,Caiji Gao

Abstract: Proximity labeling (PL), a recently developed technique to detect protein-protein interactions and subcellular structural proteomes in living cells, has been successfully applied in various animal and plant systems. Proximity labeling is conducted by fusing an engineered enzyme with catalytic activity to a protein of interest (bait protein). With the catalysis of the enzyme, small molecular substrates such as biotin are covalently linked to endogenous proximal proteins, which can be further enriched and analyzed to identify the interactome of the bait protein. TurboID, a biotin ligase produced by directed evolution, has the advantages of non-toxicity and high catalytic efficiency. Using TurboID-based proximity labeling to analyze proximal proteins of bait proteins, we can study transient or weak protein interactions, which helps to understand the complex biological processes occurring inside cells. Here, we describe methods and related tips for TurboID-based proximal labeling in Arabidopsis thaliana, and hope to provide a reference for studying plant protein-protein interactions.

Key words: TurboID, proximity labeling, biotin, protein interactions